National Library of Energy BETA

Sample records for glycoside hydrolase family

  1. Strategies to reduce end-product inhibition in family 48 glycoside hydrolases

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Chen, Mo; Bu, Lintao; Alahuhta, Markus; Brunecky, Roman; Xu, Qi; Lunin, Vladimir V.; Brady, John W.; Crowley, Michael F.; Himmel, Michael E.; Bomble, Yannick J.

    2016-02-01

    Family 48 cellobiohydrolases are some of the most abundant glycoside hydrolases in nature. They are able to degrade cellulosic biomass and therefore serve as good enzyme candidates for biofuel production. Family 48 cellulases hydrolyze cellulose chains via a processive mechanism, and produce end products composed primarily of cellobiose as well as other cellooligomers (dp ≤ 4). The challenge of utilizing cellulases in biofuel production lies in their extremely slow turnover rate. A factor contributing to the low enzyme activity is suggested to be product binding to enzyme and the resulting performance inhibition. In this study, we quantitatively evaluated the productmore » inhibitory effect of four family 48 glycoside hydrolases using molecular dynamics simulations and product expulsion free-energy calculations. We also suggested a series of single mutants of the four family 48 glycoside hydrolases with theoretically reduced level of product inhibition. As a result, the theoretical calculations provide a guide for future experimental studies designed to produce mutant cellulases with enhanced activity.« less

  2. Variants of glycoside hydrolases

    DOE Patents [OSTI]

    Teter, Sarah; Ward, Connie; Cherry, Joel; Jones, Aubrey; Harris, Paul; Yi, Jung

    2013-02-26

    The present invention relates to variants of a parent glycoside hydrolase, comprising a substitution at one or more positions corresponding to positions 21, 94, 157, 205, 206, 247, 337, 350, 373, 383, 438, 455, 467, and 486 of amino acids 1 to 513 of SEQ ID NO: 2, and optionally further comprising a substitution at one or more positions corresponding to positions 8, 22, 41, 49, 57, 113, 193, 196, 226, 227, 246, 251, 255, 259, 301, 356, 371, 411, and 462 of amino acids 1 to 513 of SEQ ID NO: 2 a substitution at one or more positions corresponding to positions 8, 22, 41, 49, 57, 113, 193, 196, 226, 227, 246, 251, 255, 259, 301, 356, 371, 411, and 462 of amino acids 1 to 513 of SEQ ID NO: 2, wherein the variants have glycoside hydrolase activity. The present invention also relates to nucleotide sequences encoding the variant glycoside hydrolases and to nucleic acid constructs, vectors, and host cells comprising the nucleotide sequences.

  3. Variants of glycoside hydrolases

    DOE Patents [OSTI]

    Teter, Sarah; Ward, Connie; Cherry, Joel; Jones, Aubrey; Harris, Paul; Yi, Jung

    2011-04-26

    The present invention relates to variants of a parent glycoside hydrolase, comprising a substitution at one or more positions corresponding to positions 21, 94, 157, 205, 206, 247, 337, 350, 373, 383, 438, 455, 467, and 486 of amino acids 1 to 513 of SEQ ID NO: 2, and optionally further comprising a substitution at one or more positions corresponding to positions 8, 22, 41, 49, 57, 113, 193, 196, 226, 227, 246, 251, 255, 259, 301, 356, 371, 411, and 462 of amino acids 1 to 513 of SEQ ID NO: 2 a substitution at one or more positions corresponding to positions 8, 22, 41, 49, 57, 113, 193, 196, 226, 227, 246, 251, 255, 259, 301, 356, 371, 411, and 462 of amino acids 1 to 513 of SEQ ID NO: 2, wherein the variants have glycoside hydrolase activity. The present invention also relates to nucleotide sequences encoding the variant glycoside hydrolases and to nucleic acid constructs, vectors, and host cells comprising the nucleotide sequences.

  4. Differential Recognition and Hydrolysis of Host Carbohydrate Antigens by Streptococcus pneumoniae Family 98 Glycoside Hydrolases

    SciTech Connect (OSTI)

    Higgins, M.; Whitworth, G; El Warry, N; Randriantsoa, M; Samain, E; Burke, R; Vocadlo, D; Boraston, A

    2009-01-01

    The presence of a fucose utilization operon in the Streptococcus pneumoniae genome and its established importance in virulence indicates a reliance of this bacterium on the harvesting of host fucose-containing glycans. The identities of these glycans, however, and how they are harvested is presently unknown. The biochemical and high resolution x-ray crystallographic analysis of two family 98 glycoside hydrolases (GH98s) from distinctive forms of the fucose utilization operon that originate from different S. pneumoniae strains reveal that one enzyme, the predominant type among pneumococcal isolates, has a unique endo-{beta}-galactosidase activity on the LewisY antigen. Altered active site topography in the other species of GH98 enzyme tune its endo-{beta}-galactosidase activity to the blood group A and B antigens. Despite their different specificities, these enzymes, and by extension all family 98 glycoside hydrolases, use an inverting catalytic mechanism. Many bacterial and viral pathogens exploit host carbohydrate antigens for adherence as a precursor to colonization or infection. However, this is the first evidence of bacterial endoglycosidase enzymes that are known to play a role in virulence and are specific for distinct host carbohydrate antigens. The strain-specific distribution of two distinct types of GH98 enzymes further suggests that S. pneumoniae strains may specialize to exploit host-specific antigens that vary from host to host, a factor that may feature in whether a strain is capable of colonizing a host or establishing an invasive infection.

  5. Targeted discovery of glycoside hydrolases from a switchgrass-adapted compost community

    SciTech Connect (OSTI)

    Allgaier, M.; Reddy, A.; Park, J. I.; Ivanova, N.; D'haeseleer, P.; Lowry, S.; Sapra, R.; Hazen, T.C.; Simmons, B.A.; VanderGheynst, J. S.; Hugenholtz, P.

    2009-11-15

    Development of cellulosic biofuels from non-food crops is currently an area of intense research interest. Tailoring depolymerizing enzymes to particular feedstocks and pretreatment conditions is one promising avenue of research in this area. Here we added a green-waste compost inoculum to switchgrass (Panicum virgatum) and simulated thermophilic composting in a bioreactor to select for a switchgrass-adapted community and to facilitate targeted discovery of glycoside hydrolases. Small-subunit (SSU) rRNA-based community profiles revealed that the microbial community changed dramatically between the initial and switchgrass-adapted compost (SAC) with some bacterial populations being enriched over 20-fold. We obtained 225 Mbp of 454-titanium pyrosequence data from the SAC community and conservatively identified 800 genes encoding glycoside hydrolase domains that were biased toward depolymerizing grass cell wall components. Of these, {approx}10% were putative cellulases mostly belonging to families GH5 and GH9. We synthesized two SAC GH9 genes with codon optimization for heterologous expression in Escherichia coli and observed activity for one on carboxymethyl cellulose. The active GH9 enzyme has a temperature optimum of 50 C and pH range of 5.5 to 8 consistent with the composting conditions applied. We demonstrate that microbial communities adapt to switchgrass decomposition using simulated composting condition and that full-length genes can be identified from complex metagenomic sequence data, synthesized and expressed resulting in active enzyme.

  6. Targeted Discovery of Glycoside Hydrolases from a Switchgrass-Adapted Compost Community

    SciTech Connect (OSTI)

    Reddy, Amitha; Allgaier, Martin; Park, Joshua I.; Ivanoval, Natalia; Dhaeseleer, Patrik; Lowry, Steve; Sapra, Rajat; Hazen, Terry C.; Simmons, Blake A.; VanderGheynst, Jean S.; Hugenholtz, Philip

    2011-05-11

    Development of cellulosic biofuels from non-food crops is currently an area of intense research interest. Tailoring depolymerizing enzymes to particular feedstocks and pretreatment conditions is one promising avenue of research in this area. Here we added a green-waste compost inoculum to switchgrass (Panicum virgatum) and simulated thermophilic composting in a bioreactor to select for a switchgrass-adapted community and to facilitate targeted discovery of glycoside hydrolases. Smallsubunit (SSU) rRNA-based community profiles revealed that the microbial community changed dramatically between the initial and switchgrass-adapted compost (SAC) with some bacterial populations being enriched over 20-fold. We obtained 225 Mbp of 454-titanium pyrosequence data from the SAC community and conservatively identified 800 genes encoding glycoside hydrolase domains that were biased toward depolymerizing grass cell wall components. Of these, ,10percent were putative cellulasesmostly belonging to families GH5 and GH9. We synthesized two SAC GH9 genes with codon optimization for heterologous expression in Escherichia coli and observed activity for one on carboxymethyl cellulose. The active GH9 enzyme has a temperature optimum of 50uC and pH range of 5.5 to 8 consistent with the composting conditions applied. We demonstrate that microbial communities adapt to switchgrass decomposition using simulated composting condition and that full-length genes can be identified from complex metagenomic sequence data, synthesized and expressed resulting in active enzyme.

  7. New perspective on glycoside hydrolase binding to lignin from pretreated corn stover

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Yarbrough, John M.; Mittal, Ashutosh; Mansfield, Elisabeth; Taylor, II, Larry E.; Hobdey, Sarah E.; Sammond, Deanne W.; Bomble, Yannick J.; Crowley, Michael F.; Decker, Stephen R.; Himmel, Michael E.; et al

    2015-12-18

    In this study, non-specific binding of cellulases to lignin has been implicated as a major factor in the loss of cellulase activity during biomass conversion to sugars. It is believed that this binding may strongly impact process economics through loss of enzyme activities during hydrolysis and enzyme recycling scenarios. The current model suggests glycoside hydrolase activities are lost though non-specific/non-productive binding of carbohydrate-binding domains to lignin, limiting catalytic site access to the carbohydrate components of the cell wall.

  8. Strategies to reduce end-product inhibition in family 48 glycoside...

    Office of Scientific and Technical Information (OSTI)

    Subject: 09 BIOMASS FUELS; 59 BASIC BIOLOGICAL SCIENCES glycoside hydrolases; product inhibition; biofuels; cellulose; molecular dynamics Word Cloud More Like This Free Publicly ...

  9. The Structure and Function of an Arabinan-specific [alpha]-1,2-Arabinofuranosidase Identified from Screening the Activities of Bacterial GH43 Glycoside Hydrolases

    SciTech Connect (OSTI)

    Cartmell, Alan; McKee, Lauren S.; Pena, Maria J.; Larsbrink, Johan; Brumer, Harry; Kaneko, Satoshi; Ichinose, Hitomi; Lewis, Richard J.; Vikso-Nielsen, Anders; Gilbert, Harry; Marles-Wright, Jon

    2012-03-26

    Reflecting the diverse chemistry of plant cell walls, microorganisms that degrade these composite structures synthesize an array of glycoside hydrolases. These enzymes are organized into sequence-, mechanism-, and structure-based families. Genomic data have shown that several organisms that degrade the plant cell wall contain a large number of genes encoding family 43 (GH43) glycoside hydrolases. Here we report the biochemical properties of the GH43 enzymes of a saprophytic soil bacterium, Cellvibrio japonicus, and a human colonic symbiont, Bacteroides thetaiotaomicron. The data show that C. japonicus uses predominantly exo-acting enzymes to degrade arabinan into arabinose, whereas B. thetaiotaomicron deploys a combination of endo- and side chain-cleaving glycoside hydrolases. Both organisms, however, utilize an arabinan-specific {alpha}-1,2-arabinofuranosidase in the degradative process, an activity that has not previously been reported. The enzyme can cleave {alpha}-1,2-arabinofuranose decorations in single or double substitutions, the latter being recalcitrant to the action of other arabinofuranosidases. The crystal structure of the C. japonicus arabinan-specific {alpha}-1,2-arabinofuranosidase, CjAbf43A, displays a five-bladed {beta}-propeller fold. The specificity of the enzyme for arabinan is conferred by a surface cleft that is complementary to the helical backbone of the polysaccharide. The specificity of CjAbf43A for {alpha}-1,2-L-arabinofuranose side chains is conferred by a polar residue that orientates the arabinan backbone such that O2 arabinose decorations are directed into the active site pocket. A shelflike structure adjacent to the active site pocket accommodates O3 arabinose side chains, explaining how the enzyme can target O2 linkages that are components of single or double substitutions.

  10. Colloid-based multiplexed method for screening plant biomass-degrading glycoside hydrolase activities in microbial communities

    SciTech Connect (OSTI)

    Reindl, W.; Deng, K.; Gladden, J.M.; Cheng, G.; Wong, A.; Singer, S.W.; Singh, S.; Lee, J.-C.; Yao, J.-S.; Hazen, T.C.; Singh, A.K; Simmons, B.A.; Adams, P.D.; Northen, T.R.

    2011-05-01

    The enzymatic hydrolysis of long-chain polysaccharides is a crucial step in the conversion of biomass to lignocellulosic biofuels. The identification and characterization of optimal glycoside hydrolases is dependent on enzyme activity assays, however existing methods are limited in terms of compatibility with a broad range of reaction conditions, sample complexity, and especially multiplexity. The method we present is a multiplexed approach based on Nanostructure-Initiator Mass Spectrometry (NIMS) that allowed studying several glycolytic activities in parallel under diverse assay conditions. Although the substrate analogs carried a highly hydrophobic perfluorinated tag, assays could be performed in aqueous solutions due colloid formation of the substrate molecules. We first validated our method by analyzing known {beta}-glucosidase and {beta}-xylosidase activities in single and parallel assay setups, followed by the identification and characterization of yet unknown glycoside hydrolase activities in microbial communities.

  11. Identifying the ionically bound cell wall and intracellular glycoside hydrolases in late growth stage Arabidopsis stems: Implications for the genetic engineering of bioenergy crops

    SciTech Connect (OSTI)

    Wei, Hui; Brunecky, Roman; Donohoe, Bryon S.; Ding, Shi -You; Ciesielski, Peter N.; Yang, Shihui; Tucker, Melvin P.; Himmel, Michael E.

    2015-05-13

    Identifying the cell wall-ionically bound glycoside hydrolases (GHs) in Arabidopsis stems is important for understanding the regulation of cell wall integrity. For cell wall proteomics studies, the preparation of clean cell wall fractions is a challenge since cell walls constitute an open compartment, which is more likely to contain a mixture of intracellular and extracellular proteins due to cell leakage at the late growth stage. Here, for this study, we utilize a CaCl2-extraction procedure to isolate non-structural proteins from Arabidopsis whole stems, followed by the in-solution and in-gel digestion methods coupled with Nano-LC-MS/MS, bioinformatics and literature analyses. This has led to the identification of 75 proteins identified using the in-solution method and 236 proteins identified by the in-gel method, among which about 10% of proteins predicted to be secreted. Together, eight cell wall proteins, namely AT1G75040, AT5G26000, AT3G57260, AT4G21650, AT3G52960, AT3G49120, AT5G49360, and AT3G14067, were identified by the in-solution method; among them, three were the GHs (AT5G26000, myrosinase 1, GH1; AT3G57260, β-1,3-glucanase 2, GH17; AT5G49360, bifunctional XYL 1/α-L-arabinofuranosidase, GH3). Moreover, four more GHs: AT4G30270 (xyloglucan endotransferase, GH16), AT1G68560 (bifunctional α-l-arabinofuranosidase/XYL, GH31), AT1G12240 (invertase, GH32) and AT2G28470 (β-galactosidase 8, GH35), were identified by the in-gel solution method only. Notably, more than half of above identified GHs are xylan- or hemicellulose-modifying enzymes, and will likely have an impact on cellulose accessibility, which is a critical factor for downstream enzymatic hydrolysis of plant tissues for biofuels production. Finally, the implications of these cell wall proteins identified at the late growth stage for the genetic engineering of bioenergy crops are discussed.

  12. Identifying the ionically bound cell wall and intracellular glycoside hydrolases in late growth stage Arabidopsis stems: Implications for the genetic engineering of bioenergy crops

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Wei, Hui; Brunecky, Roman; Donohoe, Bryon S.; Ding, Shi -You; Ciesielski, Peter N.; Yang, Shihui; Tucker, Melvin P.; Himmel, Michael E.

    2015-05-13

    Identifying the cell wall-ionically bound glycoside hydrolases (GHs) in Arabidopsis stems is important for understanding the regulation of cell wall integrity. For cell wall proteomics studies, the preparation of clean cell wall fractions is a challenge since cell walls constitute an open compartment, which is more likely to contain a mixture of intracellular and extracellular proteins due to cell leakage at the late growth stage. Here, for this study, we utilize a CaCl2-extraction procedure to isolate non-structural proteins from Arabidopsis whole stems, followed by the in-solution and in-gel digestion methods coupled with Nano-LC-MS/MS, bioinformatics and literature analyses. This has ledmore » to the identification of 75 proteins identified using the in-solution method and 236 proteins identified by the in-gel method, among which about 10% of proteins predicted to be secreted. Together, eight cell wall proteins, namely AT1G75040, AT5G26000, AT3G57260, AT4G21650, AT3G52960, AT3G49120, AT5G49360, and AT3G14067, were identified by the in-solution method; among them, three were the GHs (AT5G26000, myrosinase 1, GH1; AT3G57260, β-1,3-glucanase 2, GH17; AT5G49360, bifunctional XYL 1/α-L-arabinofuranosidase, GH3). Moreover, four more GHs: AT4G30270 (xyloglucan endotransferase, GH16), AT1G68560 (bifunctional α-l-arabinofuranosidase/XYL, GH31), AT1G12240 (invertase, GH32) and AT2G28470 (β-galactosidase 8, GH35), were identified by the in-gel solution method only. Notably, more than half of above identified GHs are xylan- or hemicellulose-modifying enzymes, and will likely have an impact on cellulose accessibility, which is a critical factor for downstream enzymatic hydrolysis of plant tissues for biofuels production. Finally, the implications of these cell wall proteins identified at the late growth stage for the genetic engineering of bioenergy crops are discussed.« less

  13. A constitutive expression system for glycosyl hydrolase family 7 cellobiohydrolases in Hypocrea jecorina

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Linger, Jeffrey G.; Taylor, II, Larry E.; Baker, John O.; Vander Wall, Todd; Hobdey, Sarah E.; Podkaminer, Kara; Himmel, Michael E.; Decker, Stephen R.

    2015-03-18

    One of the primary industrial-scale cellulase producers is the ascomycete fungus, Hypocrea jecorina, which produces and secretes large quantities of diverse cellulolytic enzymes. Perhaps the single most important biomass degrading enzyme is cellobiohydrolase I (cbh1or Cel7A) due to its enzymatic proficiency in cellulose depolymerization. However, production of Cel7A with native-like properties from heterologous expression systems has proven difficult. In this study, we develop a protein expression system in H. jecorina (Trichoderma reesei) useful for production and secretion of heterologous cellobiohydrolases from glycosyl hydrolase family 7. Building upon previous work in heterologous protein expression in filamentous fungi, we have integrated amore » native constitutive enolase promoter with the native cbh1 signal sequence. The results are the following: The constitutive eno promoter driving the expression of Cel7A allows growth on glucose and results in repression of the native cellulase system, severely reducing background endo- and other cellulase activity and greatly simplifying purification of the recombinant protein. Coupling this system to a Δcbh1 strain of H. jecorina ensures that only the recombinant Cel7A protein is produced. Two distinct transformant colony morphologies were observed and correlated with high and null protein production. Production levels in ‘fast’ transformants are roughly equivalent to those in the native QM6a strain of H. jecorina, typically in the range of 10 to 30 mg/L when grown in continuous stirred-tank fermenters. ‘Slow’ transformants showed no evidence of Cel7A production. Specific activity of the purified recombinant Cel7A protein is equivalent to that of native protein when assayed on pretreated corn stover, as is the thermal stability and glycosylation level. Purified Cel7A produced from growth on glucose demonstrated remarkably consistent specific activity. Purified Cel7A from the same strain grown on lactose

  14. Comparison of pectin-degrading fungal communities in temperate forests using glycosyl hydrolase family 28 pectinase primers targeting Ascomycete fungi

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Gacura, Matthew D.; Sprockett, Daniel D.; Heidenreich, Bess; Blackwood, Christopher B.

    2016-02-17

    Here, fungi have developed a wide assortment of enzymes to break down pectin, a prevalent polymer in plant cell walls that is important in plant defense and structure. One enzyme family used to degrade pectin is the glycosyl hydrolase family 28 (GH28). In this studywe developed primers for the amplification of GH28 coding genes from a database of 293 GH28 sequences from40 fungal genomes. The primerswere used to successfully amplify GH28 pectinases from all Ascomycota cultures tested, but only three out of seven Basidiomycota cultures. In addition, we further tested the primers in PCRs on metagenomic DNA extracted from senescedmore » tree leaves from different forest ecosystems, followed by cloning and sequencing. Taxonomic specificity for Ascomycota GH28 genes was tested by comparing GH28 composition in leaves to internal transcribed spacer (ITS) amplicon composition using pyrosequencing. All sequences obtained from GH28 primers were classified as Ascomycota; in contrast, ITS sequences indicated that fungal communitieswere up to 39% Basidiomycetes. Analysis of leaf samples indicated that both forest stand and ecosystemtype were important in structuring fungal communities. However, site played the prominent role in explaining GH28 composition, whereas ecosystem type was more important for ITS composition, indicating possible genetic drift between populations of fungi. Overall, these primers will have utility in understanding relationships between fungal community composition and ecosystem processes, as well as detection of potentially pathogenic Ascomycetes.« less

  15. On the protonation states, hydrogen bonding and catalytic mechanism of family 11 glycosidases: Direct visualization with neutrons

    SciTech Connect (OSTI)

    Fisher, Suzanne Zoe; Graham, David E.; Hanson, Leif; Kovalevskyi, Andrii Y.; Langan, Paul; Parks, Jerry M.; Wan, Qun; Ostermann, Andreas; Schrader, Tobias

    2015-10-06

    Most enzymatic reactions involve hydrogen or proton transfer among the enzyme, substrate, and water at physiological pH. Thus, enzyme catalysis cannot be fully understood without accurate mapping of hydrogen atom positions in these macromolecular catalysts. Direct information on the location of hydrogen atoms can be obtained using neutron crystallography. We used neutron crystallography and biomolecular simulation to characterize the initial stage of the glycoside hydrolysis reaction catalyzed by a family 11 glycoside hydrolase. We provide evidence that the catalytic glutamate residue alternates between two conformations bearing different basicities, first to obtain a proton from the bulk solvent, and then to deliver it to the glycosidic oxygen to initiate the hydrolysis reaction.

  16. A constitutive expression system for glycosyl hydrolase family 7 cellobiohydrolases in Hypocrea jecorina

    SciTech Connect (OSTI)

    Linger, Jeffrey G.; Taylor, II, Larry E.; Baker, John O.; Vander Wall, Todd; Hobdey, Sarah E.; Podkaminer, Kara; Himmel, Michael E.; Decker, Stephen R.

    2015-03-18

    One of the primary industrial-scale cellulase producers is the ascomycete fungus, Hypocrea jecorina, which produces and secretes large quantities of diverse cellulolytic enzymes. Perhaps the single most important biomass degrading enzyme is cellobiohydrolase I (cbh1or Cel7A) due to its enzymatic proficiency in cellulose depolymerization. However, production of Cel7A with native-like properties from heterologous expression systems has proven difficult. In this study, we develop a protein expression system in H. jecorina (Trichoderma reesei) useful for production and secretion of heterologous cellobiohydrolases from glycosyl hydrolase family 7. Building upon previous work in heterologous protein expression in filamentous fungi, we have integrated a native constitutive enolase promoter with the native cbh1 signal sequence. The results are the following: The constitutive eno promoter driving the expression of Cel7A allows growth on glucose and results in repression of the native cellulase system, severely reducing background endo- and other cellulase activity and greatly simplifying purification of the recombinant protein. Coupling this system to a Δcbh1 strain of H. jecorina ensures that only the recombinant Cel7A protein is produced. Two distinct transformant colony morphologies were observed and correlated with high and null protein production. Production levels in ‘fast’ transformants are roughly equivalent to those in the native QM6a strain of H. jecorina, typically in the range of 10 to 30 mg/L when grown in continuous stirred-tank fermenters. ‘Slow’ transformants showed no evidence of Cel7A production. Specific activity of the purified recombinant Cel7A protein is equivalent to that of native protein when assayed on pretreated corn stover, as is the thermal stability and glycosylation level. Purified Cel7A produced from growth on glucose demonstrated remarkably consistent specific activity. Purified Cel7A from the same strain grown on lactose

  17. On the protonation states, hydrogen bonding and catalytic mechanism of family 11 glycosidases: Direct visualization with neutrons

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Fisher, Suzanne Zoe; Graham, David E.; Hanson, Leif; Kovalevskyi, Andrii Y.; Langan, Paul; Parks, Jerry M.; Wan, Qun; Ostermann, Andreas; Schrader, Tobias

    2015-10-06

    Most enzymatic reactions involve hydrogen or proton transfer among the enzyme, substrate, and water at physiological pH. Thus, enzyme catalysis cannot be fully understood without accurate mapping of hydrogen atom positions in these macromolecular catalysts. Direct information on the location of hydrogen atoms can be obtained using neutron crystallography. We used neutron crystallography and biomolecular simulation to characterize the initial stage of the glycoside hydrolysis reaction catalyzed by a family 11 glycoside hydrolase. We provide evidence that the catalytic glutamate residue alternates between two conformations bearing different basicities, first to obtain a proton from the bulk solvent, and then tomore » deliver it to the glycosidic oxygen to initiate the hydrolysis reaction.« less

  18. Cellulase variants

    DOE Patents [OSTI]

    Blazej, Robert; Toriello, Nicholas; Emrich, Charles; Cohen, Richard N.; Koppel, Nitzan

    2015-07-14

    This invention provides novel variant cellulolytic enzymes having improved activity and/or stability. In certain embodiments the variant cellulotyic enzymes comprise a glycoside hydrolase with or comprising a substitution at one or more positions corresponding to one or more of residues F64, A226, and/or E246 in Thermobifida fusca Cel9A enzyme. In certain embodiments the glycoside hydrolase is a variant of a family 9 glycoside hydrolase. In certain embodiments the glycoside hydrolase is a variant of a theme B family 9 glycoside hydrolase.

  19. Community dynamics and glycoside hydrolase activities of thermophilic bacterial consortia adapted to switchgrass

    SciTech Connect (OSTI)

    Gladden, J.M.; Allgaier, M.; Miller, C.S.; Hazen, T.C.; VanderGheynst, J.S.; Hugenholtz, P.; Simmons, B.A.; Singer, S.W.

    2011-05-01

    Industrial-scale biofuel production requires robust enzymatic cocktails to produce fermentable sugars from lignocellulosic biomass. Thermophilic bacterial consortia are a potential source of cellulases and hemicellulases adapted to harsher reaction conditions than commercial fungal enzymes. Compost-derived microbial consortia were adapted to switchgrass at 60 C to develop thermophilic biomass-degrading consortia for detailed studies. Microbial community analysis using small-subunit rRNA gene amplicon pyrosequencing and short-read metagenomic sequencing demonstrated that thermophilic adaptation to switchgrass resulted in low-diversity bacterial consortia with a high abundance of bacteria related to thermophilic paenibacilli, Rhodothermus marinus, and Thermus thermophilus. At lower abundance, thermophilic Chloroflexi and an uncultivated lineage of the Gemmatimonadetes phylum were observed. Supernatants isolated from these consortia had high levels of xylanase and endoglucanase activities. Compared to commercial enzyme preparations, the endoglucanase enzymes had a higher thermotolerance and were more stable in the presence of 1-ethyl-3-methylimidazolium acetate ([C2mim][OAc]), an ionic liquid used for biomass pretreatment. The supernatants were used to saccharify [C2mim][OAc]-pretreated switchgrass at elevated temperatures (up to 80 C), demonstrating that these consortia are an excellent source of enzymes for the development of enzymatic cocktails tailored to more extreme reaction conditions.

  20. Computational Investigation of Glycosylation Effects on a Family 1 Carbohydrate-Binding Module

    SciTech Connect (OSTI)

    Taylor, C. B.; Talib, M. F.; McCabe, C.; Bu, L.; Adney, W. S.; Himmel, M. E.; Crowley, M. F.; Beckham, G. T.

    2012-01-27

    Carbohydrate-binding modules (CBMs) are ubiquitous components of glycoside hydrolases, which degrade polysaccharides in nature. CBMs target specific polysaccharides, and CBM binding affinity to cellulose is known to be proportional to cellulase activity, such that increasing binding affinity is an important component of performance improvement. To ascertain the impact of protein and glycan engineering on CBM binding, we use molecular simulation to quantify cellulose binding of a natively glycosylated Family 1 CBM. To validate our approach, we first examine aromatic-carbohydrate interactions on binding, and our predictions are consistent with previous experiments, showing that a tyrosine to tryptophan mutation yields a 2-fold improvement in binding affinity. We then demonstrate that enhanced binding of 3-6-fold over a nonglycosylated CBM is achieved by the addition of a single, native mannose or a mannose dimer, respectively, which has not been considered previously. Furthermore, we show that the addition of a single, artificial glycan on the anterior of the CBM, with the native, posterior glycans also present, can have a dramatic impact on binding affinity in our model, increasing it up to 140-fold relative to the nonglycosylated CBM. These results suggest new directions in protein engineering, in that modifying glycosylation patterns via heterologous expression, manipulation of culture conditions, or introduction of artificial glycosylation sites, can alter CBM binding affinity to carbohydrates and may thus be a general strategy to enhance cellulase performance. Our results also suggest that CBM binding studies should consider the effects of glycosylation on binding and function.

  1. Crystal Structure of the Cystic Fibrosis Transmembrane Conductance Regulator Inhibitory Factor Cif Reveals Novel Active-Site Features of an Epoxide Hydrolase Virulence Factor

    SciTech Connect (OSTI)

    Bahl, C.; Morisseau, C; Bomberger, J; Stanton, B; Hammock, B; O' Toole, G; Madden, D

    2010-01-01

    Cystic fibrosis transmembrane conductance regulator (CFTR) inhibitory factor (Cif) is a virulence factor secreted by Pseudomonas aeruginosa that reduces the quantity of CFTR in the apical membrane of human airway epithelial cells. Initial sequence analysis suggested that Cif is an epoxide hydrolase (EH), but its sequence violates two strictly conserved EH motifs and also is compatible with other {alpha}/{beta} hydrolase family members with diverse substrate specificities. To investigate the mechanistic basis of Cif activity, we have determined its structure at 1.8-{angstrom} resolution by X-ray crystallography. The catalytic triad consists of residues Asp129, His297, and Glu153, which are conserved across the family of EHs. At other positions, sequence deviations from canonical EH active-site motifs are stereochemically conservative. Furthermore, detailed enzymatic analysis confirms that Cif catalyzes the hydrolysis of epoxide compounds, with specific activity against both epibromohydrin and cis-stilbene oxide, but with a relatively narrow range of substrate selectivity. Although closely related to two other classes of {alpha}/{beta} hydrolase in both sequence and structure, Cif does not exhibit activity as either a haloacetate dehalogenase or a haloalkane dehalogenase. A reassessment of the structural and functional consequences of the H269A mutation suggests that Cif's effect on host-cell CFTR expression requires the hydrolysis of an extended endogenous epoxide substrate.

  2. Thermodynamics of enzyme-catalyzed reactions. Part 3. Hydrolases

    SciTech Connect (OSTI)

    Goldberg, R.N.; Tewari, Y.B.

    1994-11-01

    Equilibrium constants and enthalpy changes for reactions catalyzed by the hydrolase class of enzymes have been compiled. For each reaction the following information is given: the reference for the data; the reaction studied; the name of the enzyme used and its Enzyme Commission number; the method of measurement; the conditions of measurement (temperature, pH, ionic strength, and the buffer(s) and cofactor(s) used); the data and an evaluation of it; and, sometimes, commentary on the data and on any corrections which have been applied to it or any calculations for which the data have been used. The data from 146 references have been examined and evaluated. Chemical Abstract Service registry numbers are given for the substances involved in these various reactions. There is a cross reference between the substances and the Enzyme Commission numbers of the enzymes used to catalyze the reactions in which the substances participate.

  3. Conformational Variability of Organophosphorus Hydrolase upon Soman and Paraoxon Binding

    SciTech Connect (OSTI)

    Gomes, Diego Eb; Lins, Roberto D.; Pascutti, Pedro G.; Lei, Chenghong; Soares, Thereza A.

    2011-12-31

    The bacterial enzyme organophosphorus hydrolase (OPH) exhibits both catalytic and substrate promiscuity. It hydrolyzes bonds in a variety of phosphotriester (P-O), phosphonothioate (P-S), phosphofluoridate (P-F) and phosphonocyanate (F-CN) compounds. However, its catalytic efficiency varies markedly for different substrates, limiting the broad-range application of OPH as catalyst in the bioremediation of pesticides and chemical war agents. In the present study, pK{sub a} calculations and multiple explicit-solvent molecular dynamics (MD) simulations were performed to characterize and contrast the structural dynamics of OPH bound to two substrates hydrolyzed with very distinct catalytic efficiencies: the nerve agent soman (O-pinacolyl-methyl-phosphonofluoridate) and the pesticide paraoxon (diethyl p-nitrophenyl phosphate). pK{sub a} calculations for the substrate-bound and unbound enzyme showed a significant pK{sub a} shift from standard values ({Delta}pK{sub a} = {+-} 3 units) for residues 254His and 275Arg. MD simulations of the doubly protonated 254His revealed a dynamic hydrogen bond network connecting the catalytic residue 301Asp via 254His to 232Asp, 233Asp, 275Arg and 235Asp, and is consistent with a previously postulated proton relay mechanism to ferry protons away from the active site with substrates that do not require activation of the leaving group. Hydrogen bonds between 301Asp and 254His were persistent in the OPH-paraoxon complex but not in the OPH-soman one, suggesting a potential role for such interaction in the more efficient hydrolysis of paraoxon over soman by OPH. These results are in line with previous mutational studies of residue 254His, which led to an increase of the catalytic efficiency of OPH over soman yet decreased its efficiency for paraoxon. In addition, comparative analysis of the molecular trajectories for OPH bound to soman and paraoxon suggests that binding of the latter facilitates the conformational transition of OPH from the

  4. Structural Studies of the Nudix GDP-mannose Hydrolase from E. coli Reveals a New Motif for Mannose Recognition

    SciTech Connect (OSTI)

    A Boto; W Xu; J Jakoncic; A Pannuri; T Romeo; M Bessman; S Gabelli; L Amzel

    2011-12-31

    The Nudix hydrolase superfamily, characterized by the presence of the signature sequence GX5EX7REUXEEXGU (where U is I, L, or V), is a well-studied family in which relations have been established between primary sequence and substrate specificity for many members. For example, enzymes that hydrolyze the diphosphate linkage of ADP-ribose are characterized by having a proline 15 amino acids C-terminal of the Nudix signature sequence. GDPMK is a Nudix enzyme that conserves this characteristic proline but uses GDP-mannose as the preferred substrate. By investigating the structure of the GDPMK alone, bound to magnesium, and bound to substrate, the structural basis for this divergent substrate specificity and a new rule was identified by which ADP-ribose pyrophosphatases can be distinguished from purine-DP-mannose pyrophosphatases from primary sequence alone. Kinetic and mutagenesis studies showed that GDPMK hydrolysis does not rely on a single glutamate as the catalytic base. Instead, catalysis is dependent on residues that coordinate the magnesium ions and residues that position the substrate properly for catalysis. GDPMK was thought to play a role in biofilm formation because of its upregulation in response to RcsC signaling; however, GDPMK knockout strains show no defect in their capacity of forming biofilms.

  5. NREL Support for a Functional Genomics Approach to Investigate Regulation of Phenolic Glycoside: Cooperative Research and Development Final Report, CRADA number CRD-07-00218

    SciTech Connect (OSTI)

    Davis, M.

    2010-07-01

    NREL and MTU collaborated on a proposal 'A Functional Genomics Approach to Investigate Regulation of Phenolic Glycoside Metabolism in Populus' funded by the National Science Foundation.

  6. Thermal tolerant exoglucanase from Acidothermus cellulolyticus

    DOE Patents [OSTI]

    Adney, William S.; Ding, Shi-You; Vinzant, Todd B.; Himmel, Michael E.; Decker, Stephen R.; McCarter, Suzanne Lantz

    2008-07-01

    The invention provides a thermal tolerant cellulase that is a member of the glycoside hydrolase family. The invention further discloses this cellulase as Gux1. Gux1 has been isolated and characterized from Acidothermus cellulolyticus. The invention further provides recombinant forms of the identified Gux1. Methods of making and using Gux1 polypeptides, including fusions, variants, and derivatives, are also disclosed.

  7. Methods of using thermal tolerant avicelase from Acidothermus cellulolyticus

    DOE Patents [OSTI]

    Adney, William S.; Vinzant, Todd B.; Ding, Shih-You; Himmel, Michael E.

    2011-04-26

    The invention provides a thermal tolerant (thermostable) cellulase, AviIII, that is a member of the glycoside hydrolase (GH) family. AviIII was isolated and characterized from Acidothermus cellulolyticus, and, like many cellulases, the disclosed polypeptide and/or its derivatives may be useful for the conversion of biomass into biofuels and chemicals.

  8. Thermal tolerant avicelase from Acidothermus cellulolyticus

    DOE Patents [OSTI]

    Ding, Shi-You; Adney, William S.; Vinzant, Todd B.; Himmel, Michael E.

    2009-05-26

    The invention provides a thermal tolerant (thermostable) cellulase, AviIII, that is a member of the glycoside hydrolase (GH) family. AviIII was isolated and characterized from Acidothermus cellulolyticus and, like many cellulases, the disclosed polypeptide and/or its derivatives may be useful for the conversion of biomass into biofuels and chemicals.

  9. Thermal tolerant avicelase from Acidothermus cellulolyticus

    DOE Patents [OSTI]

    Ding, Shi-You; Adney, William S.; Vinzant, Todd B.; Himmel, Michael E.

    2008-04-29

    The invention provides a thermal tolerant (thermostable) cellulase, AviIII, that is a member of the glycoside hydrolase (GH) family. AviIII was isolated and characterized from Acidothermus cellulolyticus and, like many cellulases, the disclosed polypeptide and/or its derivatives may be useful for the conversion of biomass into biofuels and chemicals.

  10. Determination of Organophosphate Pesticides at a Carbon Nanotube/Organophosphorus Hydrolase Electrochemical Biosensor

    SciTech Connect (OSTI)

    Deo, R P.; Wang, Joseph; Block, I; Mulchandani, Ashok; Joshi, K; Trojanowicz, M; Scholz, F; Chen, Wilfred; Lin, Yuehe

    2005-02-08

    An amperometric biosensor for organophosphorus (OP) pesticides based on a carbon-nanotube (CNT) modified transducer and an organophosphorus hydrolase (OPH) biocatalyst is described. A bilayer approach with the OPH layer atop of the CNT film was used for preparing the CNT/OPH biosensor. The CNT layer leads to a greatly improved anodic detection of the enzymatically-generated p-nitrophenol product, including higher sensitivity and stability. The sensor performance was optimized with respect to the surface modification and operating conditions. Under the optimal conditions the biosensor was used to measure as low as 0.15 {micro}M paraoxon and 0.8 {micro}M methyl parathion with sensitivities of 25 and 6 nA/{micro}M, respectively.

  11. Insights into Substrate Specificity of NlpC/P60 Cell Wall Hydrolases Containing Bacterial SH3 Domains

    SciTech Connect (OSTI)

    Xu, Qingping; Mengin-Lecreulx, Dominique; Liu, Xueqian W.; Patin, Delphine; Farr, Carol L.; Grant, Joanna C.; Chiu, Hsiu-Ju; Jaroszewski, Lukasz; Knuth, Mark W.; Godzik, Adam; Lesley, Scott A.; Elsliger, Marc-André; Deacon, Ashley M.; Wilson, Ian A.

    2015-09-15

    ABSTRACT

    Bacterial SH3 (SH3b) domains are commonly fused with papain-like Nlp/P60 cell wall hydrolase domains. To understand how the modular architecture of SH3b and NlpC/P60 affects the activity of the catalytic domain, three putative NlpC/P60 cell wall hydrolases were biochemically and structurally characterized. These enzymes all have γ-d-Glu-A2pm (A2pm is diaminopimelic acid) cysteine amidase (ordl-endopeptidase) activities but with different substrate specificities. One enzyme is a cell wall lysin that cleaves peptidoglycan (PG), while the other two are cell wall recycling enzymes that only cleave stem peptides with an N-terminall-Ala. Their crystal structures revealed a highly conserved structure consisting of two SH3b domains and a C-terminal NlpC/P60 catalytic domain, despite very low sequence identity. Interestingly, loops from the first SH3b domain dock into the ends of the active site groove of the catalytic domain, remodel the substrate binding site, and modulate substrate specificity. Two amino acid differences at the domain interface alter the substrate binding specificity in favor of stem peptides in recycling enzymes, whereas the SH3b domain may extend the peptidoglycan binding surface in the cell wall lysins. Remarkably, the cell wall lysin can be converted into a recycling enzyme with a single mutation.

    IMPORTANCEPeptidoglycan is a meshlike polymer that envelops the bacterial plasma membrane and bestows structural integrity. Cell wall lysins and recycling enzymes are part of a set of lytic enzymes that target covalent bonds connecting the amino acid and amino sugar building blocks of the PG network. These hydrolases are involved in processes such as cell growth and division, autolysis, invasion, and PG turnover and recycling. To avoid cleavage of unintended substrates, these enzymes have very selective substrate specificities. Our biochemical and structural

  12. Insights into substrate specificity of NlpC/P60 cell wall hydrolases containing bacterial SH3 domains

    SciTech Connect (OSTI)

    Xu, Qingping; Mengin-Lecreulx, Dominique; Liu, Xueqian W.; Patin, Delphine; Farr, Carol L.; Grant, Joanna C.; Chiu, Hsiu -Ju; Jaroszewski, Lukasz; Knuth, Mark W.; Godzik, Adam; Lesley, Scott A.; Elsliger, Marc -André; Deacon, Ashley M.; Wilson, Ian A.

    2015-09-15

    Bacterial SH3 (SH3b) domains are commonly fused with papain-like Nlp/P60 cell wall hydrolase domains. To understand how the modular architecture of SH3b and NlpC/P60 affects the activity of the catalytic domain, three putative NlpC/P60 cell wall hydrolases were biochemically and structurally characterized. In addition, these enzymes all have γ-d-Glu-A2pm (A2pm is diaminopimelic acid) cysteine amidase (ordl-endopeptidase) activities but with different substrate specificities. One enzyme is a cell wall lysin that cleaves peptidoglycan (PG), while the other two are cell wall recycling enzymes that only cleave stem peptides with an N-terminall-Ala. Their crystal structures revealed a highly conserved structure consisting of two SH3b domains and a C-terminal NlpC/P60 catalytic domain, despite very low sequence identity. Interestingly, loops from the first SH3b domain dock into the ends of the active site groove of the catalytic domain, remodel the substrate binding site, and modulate substrate specificity. Two amino acid differences at the domain interface alter the substrate binding specificity in favor of stem peptides in recycling enzymes, whereas the SH3b domain may extend the peptidoglycan binding surface in the cell wall lysins. Remarkably, the cell wall lysin can be converted into a recycling enzyme with a single mutation.

    Peptidoglycan is a meshlike polymer that envelops the bacterial plasma membrane and bestows structural integrity. Cell wall lysins and recycling enzymes are part of a set of lytic enzymes that target covalent bonds connecting the amino acid and amino sugar building blocks of the PG network. These hydrolases are involved in processes such as cell growth and division, autolysis, invasion, and PG turnover and recycling. To avoid cleavage of unintended substrates, these enzymes have very selective substrate specificities. Our biochemical and structural analysis of three modular NlpC/P60

  13. Insights into substrate specificity of NlpC/P60 cell wall hydrolases containing bacterial SH3 domains

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Xu, Qingping; Mengin-Lecreulx, Dominique; Liu, Xueqian W.; Patin, Delphine; Farr, Carol L.; Grant, Joanna C.; Chiu, Hsiu -Ju; Jaroszewski, Lukasz; Knuth, Mark W.; Godzik, Adam; et al

    2015-09-15

    Bacterial SH3 (SH3b) domains are commonly fused with papain-like Nlp/P60 cell wall hydrolase domains. To understand how the modular architecture of SH3b and NlpC/P60 affects the activity of the catalytic domain, three putative NlpC/P60 cell wall hydrolases were biochemically and structurally characterized. In addition, these enzymes all have γ-d-Glu-A2pm (A2pm is diaminopimelic acid) cysteine amidase (ordl-endopeptidase) activities but with different substrate specificities. One enzyme is a cell wall lysin that cleaves peptidoglycan (PG), while the other two are cell wall recycling enzymes that only cleave stem peptides with an N-terminall-Ala. Their crystal structures revealed a highly conserved structure consisting ofmore » two SH3b domains and a C-terminal NlpC/P60 catalytic domain, despite very low sequence identity. Interestingly, loops from the first SH3b domain dock into the ends of the active site groove of the catalytic domain, remodel the substrate binding site, and modulate substrate specificity. Two amino acid differences at the domain interface alter the substrate binding specificity in favor of stem peptides in recycling enzymes, whereas the SH3b domain may extend the peptidoglycan binding surface in the cell wall lysins. Remarkably, the cell wall lysin can be converted into a recycling enzyme with a single mutation.Peptidoglycan is a meshlike polymer that envelops the bacterial plasma membrane and bestows structural integrity. Cell wall lysins and recycling enzymes are part of a set of lytic enzymes that target covalent bonds connecting the amino acid and amino sugar building blocks of the PG network. These hydrolases are involved in processes such as cell growth and division, autolysis, invasion, and PG turnover and recycling. To avoid cleavage of unintended substrates, these enzymes have very selective substrate specificities. Our biochemical and structural analysis of three modular NlpC/P60 hydrolases, one lysin, and two recycling enzymes, show

  14. Structure-Based Optimization of Arylamides as Inhibitors of Soluble Epoxide Hydrolase

    SciTech Connect (OSTI)

    Eldrup, Anne B.; Soleymanzadeh, Fariba; Taylor, Steven J.; Muegge, Ingo; Farrow, Neil A.; Joseph, David; McKellop, Keith; Man, Chuk C.; Kukulka, Alison; De Lombaert, Stephane

    2009-11-04

    Inhibition of soluble epoxide hydrolase (sEH) is hypothesized to lead to an increase in circulating levels of epoxyeicosatrienoic acids, resulting in the potentiation of their in vivo pharmacological properties. As part of an effort to identify inhibitors of sEH with high and sustained plasma exposure, we recently performed a high throughput screen of our compound collection. The screen identified N-(3,3-diphenyl-propyl)-nicotinamide as a potent inhibitor of sEH. Further profiling of this lead revealed short metabolic half-lives in microsomes and rapid clearance in the rat. Consistent with these observations, the determination of the in vitro metabolic profile of N-(3,3-diphenyl-propyl)-nicotinamide in rat liver microsomes revealed extensive oxidative metabolism and a propensity for metabolite switching. Lead optimization, guided by the analysis of the solid-state costructure of N-(3,3-diphenyl-propyl)-nicotinamide bound to human sEH, led to the identification of a class of potent and selective inhibitors. An inhibitor from this class displayed an attractive in vitro metabolic profile and high and sustained plasma exposure in the rat after oral administration.

  15. New Cellulase Identification Method Holds Promise for Lower-Cost Biofuels (Fact Sheet), NREL (National Renewable Energy Laboratory)

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    A new computational approach to genomic data effectively distinguishes cellulases and non-cellulases within the protein family GH48, a key component for degrading lignocellulose for biofuels. Cellulase enzymes, particularly from the glycoside hydrolase family 48 (GH48), are a critical component of many lignocellulose-degrading systems that produce biomass to use for biofu- els. Typically it has been difficult and costly-but necessary-to distinguish required cellulase enzymes from unwanted

  16. A thiocyanate hydrolase of Thiobacillus thioparus. A novel enzyme catalyzing the formation of carbonyl sulfide from thiocyanate

    SciTech Connect (OSTI)

    Katayama, Y.; Narahara, Y.; Inoue, Y.; Amano, F.; Kanagawa, T.; Kuraishi, H. (Faculty of Agriculture, Tokyo University of Agriculture and Technology, (Japan))

    1992-05-05

    A thiocyanate hydrolase that catalyzes the first step in thiocyanate degradation was purified to homogeneity from Thiobacillus thioparus, an obligate chemolithotrophic eubacterium metabolizing thiocyanate to sulfate as an energy source. The thiocyanate hydrolase was purified 52-fold by steps involving ammonium sulfate precipitation, DEAE-Sephacel column chromatography, and hydroxylapatite column chromatography. The enzyme hydrolyzed 1 mol of thiocyanate to form 1 mol of carbonyl sulfide and 1 mol of ammonia as follows: SCN- + 2H2O----COS + NH3 + OH-. This is the first report describing the hydrolysis of thiocyanate to carbonyl sulfide by an enzyme. The enzyme had a molecular mass of 126 kDa and was composed of three different subunits: alpha (19 kDa), beta (23 kDa), and gamma (32 kDa). The enzyme exhibited optimal activities at pH 7.5-8.0 and at temperatures ranging from 30 to 40 degrees C. The Km value for thiocyanate was approximately 11 mM. Immunoblot analysis with polyclonal antibodies against the purified enzyme suggested that it was induced in T. thioparus cells when the cells were grown with thiocyanate.

  17. Thermal Tolerant Cellulase from Acidothermus Cellulolyticus

    DOE Patents [OSTI]

    Ding, S. Y.; Adney, W. S.; Vinzant, T. B.; Himmel, M. E.; Decker, S. R.

    2006-06-13

    The invention provides a thermal tolerant cellulase that is a member of the glycoside hydrolase family. The invention further discloses this cellulase as GuxA. GuxA has been isolated and characterized from Acidothermus cellulolyticus. The invention further provides recombinant forms of the identified GuxA. Methods of making and using GuxA polypeptides, including fusions, variants, and derivatives, are also disclosed.

  18. Thermal tolerant cellulase from Acidothermus cellulolyticus

    DOE Patents [OSTI]

    Ding, Shi-You; Adney, William S.; Vinzant, Todd B.; Himmel, Michael E.; Decker, Stephen R.

    2006-06-13

    The invention provides a thermal tolerant cellulase that is a member of the glycoside hydrolase family. The invention further discloses this cellulase as GuxA. GuxA has been isolated and characterized from Acidothermus cellulolyticus. The invention further provides recombinant forms of the identified GuxA. Methods of making and using GuxA polypeptides, including fusions, variants, and derivatives, are also disclosed.

  19. Successful Family Engagement

    Broader source: Energy.gov [DOE]

    Families are a young child's most important first teachers, but engaging busy families in meaningful interactive learning can be a challenge. Join Pittsburgh's Carnegie Science Center and...

  20. Thermal tolerant mannanase from acidothermus cellulolyticus ...

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    - Visit the NREL Commercialization and Technology Transfer Website Abstract: The invention provides a thermal tolerant mannanase that is a member of the glycoside hydrolase...

  1. Thermal tolerant exoglucanase from acidothermus cellulolyticus...

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    - Visit the NREL Commercialization and Technology Transfer Website Abstract: The invention provides a thermal tolerant cellulase that is a member of the glycoside hydrolase...

  2. Thermal tolerant cellulase from acidothermus cellulolyticus ...

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    - Visit the NREL Commercialization and Technology Transfer Website Abstract: The invention provides a thermal tolerant cellulase that is a member of the glycoside hydrolase...

  3. Thermal tolerant cellulase from Acidothermus cellulolyticus ...

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Enzymes for the Conversion of Biomass to Biofuels and Chemicals Abstract: The invention provides a thermal tolerant cellulase that is a member of the glycoside hydrolase...

  4. Thermal tolerant exoglucanase from Acidothermus cellulolyticus...

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Enzymes for the Conversion of Biomass to Biofuels and Chemicals Abstract: The invention provides a thermal tolerant cellulase that is a member of the glycoside hydrolase...

  5. Thermal tolerant mannanase from acidothermus cellulolyticus ...

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Enzymes for the Conversion of Biomass to Biofuels and Chemicals Abstract: The invention provides a thermal tolerant mannanase that is a member of the glycoside hydrolase...

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    ... Filter Results Filter by Subject biomass fuels (3) basic biological sciences (1) basic biological sciences glycoside hydrolases (1) bioethanol (1) biofuels (1) cellulases (1) ...

  7. The Vital Function of Fe3O4@Au nanocomposites for Hydrolase Biosensor Design and Its Application in Detection of Methyl Parathion

    SciTech Connect (OSTI)

    Zhao, Yuting; Zhang, Weiying; Lin, Yuehe; Du, Dan

    2013-02-04

    A nanocomposite of gold nanoparticles (AuNPs) decorating a magnetic Fe3O4 core was synthesized using cysteamine (SHNH2) as linker, and characterized by TEM, XPS, UV and electrochemistry. Then a hydrolase biosensor, based on self-assembly of methyl parathion hydrolase (MPH) on the Fe3O4@Au nanocomposite, was developed for sensitive and selective detection of the organophosphorus pesticide (OP) methyl parathion. The magnetic nanocomposite provides an easy way to construct the enzyme biosensor by simply exerting an external magnetic field, and also provides a simple way to renew the electrode surface by removing the magnet. Unlike inhibition-based enzyme biosensors, the hydrolase is not poisoned by OPs and thus is reusable for continuous measurement. AuNPs not only provide a large surface area, high loading efficiency and fast electron transfer, but also stabilize the enzyme through electrostatic interactions. The MPH biosensor shows rapid response and high selectivity for detection of methyl parathion, with a linear range from 0.5 to 1000 ng/mL and a detection limit of 0.1 ng/mL. It also shows acceptable reproducibility and stability. The simplicity and ease of operation of the proposed method has great potential for on-site detection of PS containing pesticides and provides a promising strategy to construct a robust biosensor.

  8. X-ray Crystallographic Analysis of [alpha]-Ketoheterocycle Inhibitors Bound to a Humanized Variant of Fatty Acid Amide Hydrolase

    SciTech Connect (OSTI)

    Mileni, Mauro; Garfunkle, Joie; Ezzili, Cyrine; Kimball, F.Scott; Cravatt, Benjamin F.; Stevens, Raymond C.; Boger, Dale L.

    2010-11-03

    Three cocrystal X-ray structures of the {alpha}-ketoheterocycle inhibitors 3-5 bound to a humanized variant of fatty acid amide hydrolase (FAAH) are disclosed and comparatively discussed alongside those of 1 (OL-135) and its isomer 2. These five X-ray structures systematically probe each of the three active site regions key to substrate or inhibitor binding: (1) the conformationally mobile acyl chain-binding pocket and membrane access channel responsible for fatty acid amide substrate and inhibitor acyl chain binding, (2) the atypical active site catalytic residues and surrounding oxyanion hole that covalently binds the core of the {alpha}-ketoheterocycle inhibitors captured as deprotonated hemiketals mimicking the tetrahedral intermediate of the enzyme-catalyzed reaction, and (3) the cytosolic port and its uniquely important imbedded ordered water molecules and a newly identified anion binding site. The detailed analysis of their key active site interactions and their implications on the interpretation of the available structure-activity relationships are discussed providing important insights for future design.

  9. The Role of Non-Bonded Interactions in the Conformational Dynamics of Organophosphorous Hydrolase Adsorbed onto Functionalized Mesoporous Silica Surfaces

    SciTech Connect (OSTI)

    Gomes, Diego Enry B.; Lins, Roberto D.; Pascutti, Pedro G.; Lei, Chenghong; Soares, Thereza A.

    2010-01-14

    The enzyme organophosphorous hydrolase (OPH) catalyzes the hydrolysis of a wide variety of organophosphorous compounds with high catalytic efficiency and broad substrate specificity. The immobilization of OPH in functionalized mesoporous silica surfaces increases significantly its catalytic specific activity compared to the enzyme in solution with important applications for the detection and decontamination of insecticides and chemical warfare agents. Experimental measurements of immobilization efficiency as function of the charge and coverage percentage of different functional groups have been interpreted as electrostatic forces being the predominant interactions underlying the adsorption of OPH onto functionalized mesoporous silica surfaces. Explicit solvent molecular dynamics simulations have been performed for OPH in bulk solution and adsorbed onto two distinct interaction potential models of the mesoporous silica functional groups in order to investigate the relative contributions of non-bonded interactions to the conformational dynamics and adsorption of the protein. Our results support the conclusion that while electrostatic interactions are responsible for the specific binding of OPH to the FMS surface, van der Waals forces are detrimental for enhanced interfacial adhesion. Key-words: molecular dynamics simulations; bacterial phosphotriesterase; conformational changes; confined environments; coarse-grain and atomistic models; silanol molecular model.

  10. Energy Landscape for the Interaction of the Family 1 Carbohydrate-Binding Module and the Cellulose Surface is Altered by Hydrolyzed Glycosidic Bonds

    SciTech Connect (OSTI)

    Bu, L.; Beckham, G. T.; Crowley, M. F.; Chang, C. H.; Matthews, J. F.; Bomble, Y. J.; Adney, W. S.; Himmel, M. E.; Nimlos, M. R.

    2009-01-01

    A multiscale simulation model is used to construct potential and free energy surfaces for the carbohydrate-binding module [CBM] from an industrially important cellulase, Trichoderma reesei cellobiohydrolase I, on the hydrophobic face of a coarse-grained cellulose I{beta} polymorph. We predict from computation that the CBM alone exhibits regions of stability on the hydrophobic face of cellulose every 5 and 10 {angstrom}, corresponding to a glucose unit and a cellobiose unit, respectively. In addition, we predict a new role for the CBM: specifically, that in the presence of hydrolyzed cellulose chain ends, the CBM exerts a thermodynamic driving force to translate away from the free cellulose chain ends. This suggests that the CBM is not only required for binding to cellulose, as has been known for two decades, but also that it has evolved to both assist the enzyme in recognizing a cellulose chain end and exert a driving force on the enzyme during processive hydrolysis of cellulose.

  11. Development of monoclonal antibodies to human microsomal epoxide hydrolase and analysis of preneoplastic antigen-like molecules

    SciTech Connect (OSTI)

    Duan, Hongying; Yoshimura, Kazunori; Kobayashi, Nobuharu; Sugiyama, Kazuo; Sawada, Jun-ichi; Saito, Yoshiro; Morisseau, Christophe; Hammock, Bruce D.; Akatsuka, Toshitaka

    2012-04-01

    Microsomal epoxide hydrolase (mEH) is a drug metabolizing enzyme which resides on the endoplasmic reticulum (ER) membrane and catalyzes the hydration of reactive epoxide intermediates that are formed by cytochrome P450s. mEH is also thought to have a role in bile acid transport on the plasma membrane of hepatocytes. It is speculated that efficient execution of such multiple functions is secured by its orientation and association with cytochrome P450 enzymes on the ER membrane and formation of a multiple transport system on the plasma membrane. In certain disease status, mEH loses its association with the membrane and can be detected as distinct antigens in the cytosol of preneoplastic foci of liver (preneoplastic antigen), in the serum in association with hepatitis C virus infection (AN antigen), or in some brain tumors. To analyze the antigenic structures of mEH in physiological and pathological conditions, we developed monoclonal antibodies against different portions of mEH. Five different kinds of antibodies were obtained: three, anti-N-terminal portions; one anti-C-terminal; and one, anti-conformational epitope. By combining these antibodies, we developed antigen detection methods which are specific to either the membrane-bound form or the linearized form of mEH. These methods detected mEH in the culture medium released from a hepatocellular carcinoma cell line and a glioblastoma cell line, which was found to be a multimolecular complex with a unique antigenic structure different from that of the membrane-bound form of mEH. These antibodies and antigen detection methods may be useful to study pathological changes of mEH in various human diseases. -- Highlights: ? Monoclonal antibodies against different portions of mEH were developed. ? They discriminate between the membrane-bound and the linearized forms of mEH. ? We analyze the antigenic structure of the altered form of mEH in tumor cells. ? Preneoplastic antigen is a multimolecular complex of mEH with a

  12. Mutational, Structural, and Kinetic Evidence for a Dissociative Mechanism in the GDP-mannose Mannosyl Hydrolase Reaction

    SciTech Connect (OSTI)

    Xia,Z.; Azurmendi, H.; lairson, L.; Withers, S.; Gabelli, S.; Bianchet, M.; Amzel, L.; Mildvan, A.

    2005-01-01

    GDP-mannose hydrolase (GDPMH) catalyzes the hydrolysis of GDP-{alpha}-D-sugars by nucleophilic substitution with inversion at the anomeric C1 atom of the sugar, with general base catalysis by H124. Three lines of evidence indicate a mechanism with dissociative character. First, in the 1.3 Angstrom X-ray structure of the GDPMH-Mg{sup 2+}-GDP{center_dot}Tris{sup +} complex, the GDP leaving group interacts with five catalytic components: R37, Y103, R52, R65, and the essential Mg{sup 2+}. As determined by the effects of site-specific mutants on k{sub cat}, these components contribute factors of 24-, 100-, 309-, 24-, and {ge}10{sup 5}-fold, respectively, to catalysis. Both R37 and Y103 bind the {beta}-phosphate of GDP and are only 5.0 Angstroms apart. Accordingly, the R37Q/Y103F double mutant exhibits partially additive effects of the two single mutants on k{sub cat}, indicating cooperativity of R37 and Y103 in promoting catalysis, and antagonistic effects on K{sub m}. Second, the conserved residue, D22, is positioned to accept a hydrogen bond from the C2-OH group of the sugar undergoing substitution at C1, as was shown by modeling an {alpha}-D-mannosyl group into the sugar binding site. The D22A and D22N mutations decreased k{sub cat} by factors of 10{sup 2.1} and 10{sup 2.6}, respectively, for the hydrolysis of GDP-{alpha}-D-mannose, and showed smaller effects on K{sub m}, suggesting that the D22 anion stabilizes a cationic oxocarbenium transition state. Third, the fluorinated substrate, GDP-2F-{alpha}-D-mannose, for which a cationic oxocarbenium transition state would be destabilized by electron withdrawal, exhibited a 16-fold decrease in k{sub cat} and a smaller, 2.5-fold increase in K{sub m}. The D22A and D22N mutations further decreased the k{sub cat} with GDP-2F-{alpha}-D-mannose to values similar to those found with GDP-{alpha}-D-mannose, and decreased the K{sub m} of the fluorinated substrate. The choice of histidine as the general base over glutamate, the

  13. X-ray, NMR, and Mutational Studies of the Catalytic Cycle of the GDP-Mannose Mannosyl Hydrolase Reaction

    SciTech Connect (OSTI)

    Gabelli,S.; Azurmendi, H.; Bianchet, M.; Amzel, L.; Mildva, A.

    2006-01-01

    GDP-mannose hydrolase catalyzes the hydrolysis with inversion of GDP-{alpha}-D-hexose to GDP and {beta}-D-hexose by nucleophilic substitution by water at C1 of the sugar. Two new crystal structures (free enzyme and enzyme-substrate complex), NMR, and site-directed mutagenesis data, combined with the structure of the enzyme-product complex reported earlier, suggest a four-stage catalytic cycle. An important loop (L6, residues 119-125) contains a ligand to the essential Mg{sup 2+} (Gln-123), the catalytic base (His-124), and three anionic residues. This loop is not ordered in the X-ray structure of the free enzyme due to dynamic disorder, as indicated by the two-dimensional 1H-15N HMQC spectrum, which shows selective exchange broadening of the imidazole nitrogen resonances of His-124 (k{sub ex} = 6.6 x 10{sup 4} s{sup -1}). The structure of the enzyme-Mg{sup 2+}-GDP-mannose substrate complex of the less active Y103F mutant shows loop L6 in an open conformation, while the structure of the enzyme-Mg{sup 2+}-GDP product complex showed loop L6 in a closed, 'active' conformation. 1H-15N HMQC spectra show the imidazole N of His-124 to be unprotonated, appropriate for general base catalysis. Substituting Mg{sup 2+} with the more electrophilic metal ions Mn{sup 2+} or Co{sup 2+} decreases the pK{sub a} in the pH versus k{sub cat} rate profiles, showing that deprotonation of a metal-bound water is partially rate-limiting. The H124Q mutation, which decreases k{sub cat} 103.4-fold and largely abolishes its pH dependence, is rescued by the Y103F mutation, which increases k{sub cat} 23-fold and restores its pH dependence. The structural basis of the rescue is the fact that the Y103F mutation shifts the conformational equilibrium to the open form moving loop L6 out of the active site, thus permitting direct access of the specific base hydroxide from the solvent. In the proposed dissociative transition state, which occurs in the closed, active conformation of the enzyme, the

  14. Thermal tolerant mannanase from acidothermus cellulolyticus

    DOE Patents [OSTI]

    Ding, Shi-You; Adney, William S.; Vinzant, Todd B.; Himmel, Michael E.

    2006-09-26

    The invention provides a thermal tolerant mannanase that is a member of the glycoside hydrolase family. The invention further discloses this mannanase as ManA. ManA has been isolated and characterized from Acidothermus cellulolyticus. The invention further provides recombinant forms of the identified ManA. Methods of making ManA polypeptides, including fusions, variants, and derivatives, are also disclosed. Methods of using mannanase A, including for the processing of food and for use in food stuffs as bulking agents and the like, are also disclosed.

  15. The Structural Basis of Substrate Recognition in an exo-b-d-glucosaminidase Involved in Chitosan Hydrolysis

    SciTech Connect (OSTI)

    Van Bueren, A.; Ghinet, M; Gregg, K; Fleury, A; Brzezinski, R; Boraston, A

    2009-01-01

    Family 2 of the glycoside hydrolase classification is one of the largest families. Structurally characterized members of this family include enzymes with ?-galactosidase activity (Escherichia coli LacZ), ?-glucuronidase activity (Homo sapiens GusB), and ?-mannosidase activity (Bacteroides thetaiotaomicron BtMan2A). Here, we describe the structure of a family 2 glycoside hydrolase, CsxA, from Amycolatopsis orientalis that has exo-?-d-glucosaminidase (exo-chitosanase) activity. Analysis of a product complex (1.85 A resolution) reveals a unique negatively charged pocket that specifically accommodates the nitrogen of nonreducing end glucosamine residues, allowing this enzyme to discriminate between glucose and glucosamine. This also provides structural evidence for the role of E541 as the catalytic nucleophile and D469 as the catalytic acid/base. The structures of an E541A mutant in complex with a natural ?-1,4-d-glucosamine tetrasaccharide substrate and both E541A and D469A mutants in complex with a pNP-?-d-glucosaminide synthetic substrate provide insight into interactions in the + 1 subsite of this enzyme. Overall, a comparison with the active sites of other GH2 enzymes highlights the unique architecture of the CsxA active site, which imparts specificity for its cationic substrate.

  16. The Structural Basis of Substrate Recognition in an exo-beta-d-Glucosaminidase Involved in Chitosan Hydrolysis

    SciTech Connect (OSTI)

    Lammerts van Bueren, A.; Ghinet, M; Gregg, K; Fleury, A; Brzezinski, R; Boraston, A

    2009-01-01

    Family 2 of the glycoside hydrolase classification is one of the largest families. Structurally characterized members of this family include enzymes with beta-galactosidase activity (Escherichia coli LacZ), beta-glucuronidase activity (Homo sapiens GusB), and beta-mannosidase activity (Bacteroides thetaiotaomicron BtMan2A). Here, we describe the structure of a family 2 glycoside hydrolase, CsxA, from Amycolatopsis orientalis that has exo-beta-D-glucosaminidase (exo-chitosanase) activity. Analysis of a product complex (1.85 A resolution) reveals a unique negatively charged pocket that specifically accommodates the nitrogen of nonreducing end glucosamine residues, allowing this enzyme to discriminate between glucose and glucosamine. This also provides structural evidence for the role of E541 as the catalytic nucleophile and D469 as the catalytic acid/base. The structures of an E541A mutant in complex with a natural beta-1,4-D-glucosamine tetrasaccharide substrate and both E541A and D469A mutants in complex with a pNP-beta-D-glucosaminide synthetic substrate provide insight into interactions in the +1 subsite of this enzyme. Overall, a comparison with the active sites of other GH2 enzymes highlights the unique architecture of the CsxA active site, which imparts specificity for its cationic substrate.

  17. Deletion of Caldicellulosiruptor bescii CelA reveals its crucial role in the deconstruction of lignocellulosic biomass

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Young, Jenna; Chung, Daehwan; Bomble, Yannick J.; Himmel, Michael E.; Westpheling, Janet

    2014-10-09

    Background: Members of the bacterial genus Caldicellulosiruptor are the most thermophilic cellulolytic organisms described to date, and have the ability to grow on lignocellulosic biomass without conventional pretreatment. Different species vary in their abilities to degrade cellulose, and the presence of CelA, a bifunctional glycoside hydrolase that contains a Family 48 and a Family 9 catalytic domain, correlates well with cellulolytic ability in members of this genus. For example, C. hydrothermalis, which does not contain a CelA homolog, or a GH48 Family or GH9 Family glycoside hydrolase, is the least cellulolytic of the Caldicellulosiruptor species so far described. C. bescii,more » which contains CelA and expresses it constitutively, is among the most cellulolytic. In fact, CelA is the most abundant extracellular protein produced in C. bescii. The enzyme contains two catalytic units, a Family 9A-CBM3c processive endoglucanase and a Family 48 exoglucanase, joined by two Family 3b carbohydrate-binding domains. Although there are two non-reducing end-specific Family 9 and three reducing end-specific Family 48 glycoside hydrolases (producing primarily glucose and cellobiose; and cellobiose and cellotriose, respectively) in C. bescii, CelA is the only protein that combines both enzymatic activities. Results: A deletion of the celA gene resulted in a dramatic reduction in the microorganism’s ability to grow on crystalline cellulose (Avicel) and diminished growth on lignocellulosic biomass. A comparison of the overall endoglucanase and exoglucanase activities of the mutant compared with the wild-type suggests that the loss of the endoglucanase activity provided by the GH9 family domain is perhaps compensated for by other enzymes produced by the cell. In contrast, it appears that no other enzymes in the C. bescii secretome can compensate for the loss of exoglucanase activity. The change in enzymatic activity in the celA mutant resulted in a 15-fold decrease in

  18. Deletion of Caldicellulosiruptor bescii CelA reveals its crucial role in the deconstruction of lignocellulosic biomass

    SciTech Connect (OSTI)

    Young, Jenna; Chung, Daehwan; Bomble, Yannick J.; Himmel, Michael E.; Westpheling, Janet

    2014-10-09

    Background: Members of the bacterial genus Caldicellulosiruptor are the most thermophilic cellulolytic organisms described to date, and have the ability to grow on lignocellulosic biomass without conventional pretreatment. Different species vary in their abilities to degrade cellulose, and the presence of CelA, a bifunctional glycoside hydrolase that contains a Family 48 and a Family 9 catalytic domain, correlates well with cellulolytic ability in members of this genus. For example, C. hydrothermalis, which does not contain a CelA homolog, or a GH48 Family or GH9 Family glycoside hydrolase, is the least cellulolytic of the Caldicellulosiruptor species so far described. C. bescii, which contains CelA and expresses it constitutively, is among the most cellulolytic. In fact, CelA is the most abundant extracellular protein produced in C. bescii. The enzyme contains two catalytic units, a Family 9A-CBM3c processive endoglucanase and a Family 48 exoglucanase, joined by two Family 3b carbohydrate-binding domains. Although there are two non-reducing end-specific Family 9 and three reducing end-specific Family 48 glycoside hydrolases (producing primarily glucose and cellobiose; and cellobiose and cellotriose, respectively) in C. bescii, CelA is the only protein that combines both enzymatic activities. Results: A deletion of the celA gene resulted in a dramatic reduction in the microorganism’s ability to grow on crystalline cellulose (Avicel) and diminished growth on lignocellulosic biomass. A comparison of the overall endoglucanase and exoglucanase activities of the mutant compared with the wild-type suggests that the loss of the endoglucanase activity provided by the GH9 family domain is perhaps compensated for by other enzymes produced by the cell. In contrast, it appears that no other enzymes in the C. bescii secretome can compensate for the loss of exoglucanase activity. The change in enzymatic activity in the celA mutant resulted in a 15-fold decrease in sugar

  19. Family Chemistry | GE Global Research

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Family Chemistry Click to email this to a friend (Opens in new window) Share on Facebook (Opens in new window) Click to share (Opens in new window) Click to share on LinkedIn (Opens in new window) Click to share on Tumblr (Opens in new window) Family Chemistry Family gathers at the dinner table and, sometimes, the conference table or the science lab. Paul and Mark Buckley both work at the Global Research Center in Niskayuna, New York and, collectively, have more than 45 years of experience

  20. Metatranscriptomic analysis of lignocellulolytic microbial communities involved in high-solids decomposition of rice straw

    SciTech Connect (OSTI)

    Simmons, Christopher W.; Reddy, Amitha P.; D’haeseleer, Patrik; Khudyakov, Jane; Billis, Konstantinos; Pati, Amrita; Simmons, Blake A.; Singer, Steven W.; Thelen, Michael P.; VanderGheynst, Jean S.

    2014-12-31

    New lignocellulolytic enzymes are needed that maintain optimal activity under the harsh conditions present during industrial enzymatic deconstruction of biomass, including high temperatures, the absence of free water, and the presence of inhibitors from the biomass. Enriching lignocellulolytic microbial communities under these conditions provides a source of microorganisms that may yield robust lignocellulolytic enzymes tolerant to the extreme conditions needed to improve the throughput and efficiency of biomass enzymatic deconstruction. Identification of promising enzymes from these systems is challenging due to complex substrate-enzyme interactions and requirements to assay for activity. In this study, metatranscriptomes from compost-derived microbial communities enriched on rice straw under thermophilic and mesophilic conditions were sequenced and analyzed to identify lignocellulolytic enzymes overexpressed under thermophilic conditions. To determine differential gene expression across mesophilic and thermophilic treatments, a method was developed which pooled gene expression by functional category, as indicated by Pfam annotations, since microbial communities performing similar tasks are likely to have overlapping functions even if they share no specific genes. Differential expression analysis identified enzymes from glycoside hydrolase family 48, carbohydrate binding module family 2, and carbohydrate binding module family 33 domains as significantly overexpressed in the thermophilic community. Overexpression of these protein families in the thermophilic community resulted from expression of a small number of genes not currently represented in any protein database. Genes in overexpressed protein families were predominantly expressed by a single Actinobacteria genus, Micromonospora. In conclusion, coupling measurements of deconstructive activity with comparative analyses to identify overexpressed enzymes in lignocellulolytic communities provides a targeted

  1. Metatranscriptomic analysis of lignocellulolytic microbial communities involved in high-solids decomposition of rice straw

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Simmons, Christopher W.; Reddy, Amitha P.; D’haeseleer, Patrik; Khudyakov, Jane; Billis, Konstantinos; Pati, Amrita; Simmons, Blake A.; Singer, Steven W.; Thelen, Michael P.; VanderGheynst, Jean S.

    2014-12-31

    New lignocellulolytic enzymes are needed that maintain optimal activity under the harsh conditions present during industrial enzymatic deconstruction of biomass, including high temperatures, the absence of free water, and the presence of inhibitors from the biomass. Enriching lignocellulolytic microbial communities under these conditions provides a source of microorganisms that may yield robust lignocellulolytic enzymes tolerant to the extreme conditions needed to improve the throughput and efficiency of biomass enzymatic deconstruction. Identification of promising enzymes from these systems is challenging due to complex substrate-enzyme interactions and requirements to assay for activity. In this study, metatranscriptomes from compost-derived microbial communities enriched onmore » rice straw under thermophilic and mesophilic conditions were sequenced and analyzed to identify lignocellulolytic enzymes overexpressed under thermophilic conditions. To determine differential gene expression across mesophilic and thermophilic treatments, a method was developed which pooled gene expression by functional category, as indicated by Pfam annotations, since microbial communities performing similar tasks are likely to have overlapping functions even if they share no specific genes. Differential expression analysis identified enzymes from glycoside hydrolase family 48, carbohydrate binding module family 2, and carbohydrate binding module family 33 domains as significantly overexpressed in the thermophilic community. Overexpression of these protein families in the thermophilic community resulted from expression of a small number of genes not currently represented in any protein database. Genes in overexpressed protein families were predominantly expressed by a single Actinobacteria genus, Micromonospora. In conclusion, coupling measurements of deconstructive activity with comparative analyses to identify overexpressed enzymes in lignocellulolytic communities provides a

  2. The DOE Feeds Families Festival

    Broader source: Energy.gov [DOE]

    Department of Energy employees took a moment to give back to the Washington, D.C. community by holding our first "DOE Feeds Families Festival" outside of the Forrestal Building.

  3. Multi-Family Housing Loans and Grants

    Broader source: Energy.gov [DOE]

    Multi-family housing programs offer rural rental housing loans to provide affordable multi-family rental housing for very low-, low-, and moderate-income families, the elderly, and persons with...

  4. Feds feed Families | Department of Energy

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Feds feed Families Feds feed Families September 3, 2014 - 12:27pm Addthis Feds feed Families Feds feed Families September 2, 2014 Feds feed Families Representatives of the Office of Enterprise Assessments delivered more than 1,600 pounds of non-perishable food to the Manna Food Center on September 2, 2014 as part of the 2014 Feds Feed Families campaign. Manna feeds about 3,300 needy families in Montgomery County. Manna also provides food to 48 Montgomery County soup kitchens, food pantries,

  5. Product Binding Varies Dramatically between Processive and Nonprocessive Cellulase Enzymes

    SciTech Connect (OSTI)

    Bu, L.; Nimlos, M. R.; Shirts, M. R.; Stahlberg, J.; Himmel, M. E.; Crowley, M. F.; Beckham, G. T.

    2012-07-13

    Cellulases hydrolyze {beta}-1,4 glycosidic linkages in cellulose, which are among the most prevalent and stable bonds in Nature. Cellulases comprise many glycoside hydrolase families and exist as processive or nonprocessive enzymes. Product inhibition negatively impacts cellulase action, but experimental measurements of product-binding constants vary significantly, and there is little consensus on the importance of this phenomenon. To provide molecular level insights into cellulase product inhibition, we examine the impact of product binding on processive and nonprocessive cellulases by calculating the binding free energy of cellobiose to the product sites of catalytic domains of processive and nonprocessive enzymes from glycoside hydrolase families 6 and 7. The results suggest that cellobiose binds to processive cellulases much more strongly than nonprocessive cellulases. We also predict that the presence of a cellodextrin bound in the reactant site of the catalytic domain, which is present during enzymatic catalysis, has no effect on product binding in nonprocessive cellulases, whereas it significantly increases product binding to processive cellulases. This difference in product binding correlates with hydrogen bonding between the substrate-side ligand and the cellobiose product in processive cellulase tunnels and the additional stabilization from the longer tunnel-forming loops. The hydrogen bonds between the substrate- and product-side ligands are disrupted by water in nonprocessive cellulase clefts, and the lack of long tunnel-forming loops results in lower affinity of the product ligand. These findings provide new insights into the large discrepancies reported for binding constants for cellulases and suggest that product inhibition will vary significantly based on the amount of productive binding for processive cellulases on cellulose.

  6. Crystallization and preliminary diffraction studies of CBM3b of cellobiohydrolase 9A from Clostridium thermocellum

    SciTech Connect (OSTI)

    Jindou, Sadanari; Petkun, Svetlana; Shimon, Linda; Bayer, Edward A.; Lamed, Raphael; Frolow, Felix

    2007-12-01

    The cloning, expression, purification and crystallization of the CBM3b module of cellobiohydrolase 9A from C. thermocellum is described. The crystals diffract to 2.68 Å. Family 3 carbohydrate-binding modules (CBM3s) are associated with the scaffoldin subunit of the multi-enzyme cellulosome complex and with the family 9 glycoside hydrolases, which are multimodular enzymes that act on plant cell-wall polysaccharides, notably cellulose. Here, the crystallization of CBM3b from cellobiohydrolase 9A is reported. The crystals are tetragonal and belong to space group P4{sub 1} or P4{sub 3}. X-ray diffraction data for CBM3b have been collected to 2.68 Å resolution on beamline ID14-4 at the ESRF.

  7. 'Lollachilipalooza' benefits Feds Feed Families effort | National...

    National Nuclear Security Administration (NNSA)

    'Lollachilipalooza' benefits Feds Feed Families effort Deputy Secretary of Energy Daniel Poneman led his band, "Yellow Cake," at "Lollachilipalooza" today to benefit DOE's 2014 ...

  8. Proteomic and Functional Analysis of the Cellulase System Expressed by Postia placenta during Brown Rot of Solid Wood

    SciTech Connect (OSTI)

    Ryu, Jae San; Shary, Semarjit; Houtman, Carl J.; Panisko, Ellen A.; Korripally, Premsagar; St John, Franz J.; Crooks, Casey; Siika-aho, Matti; Magnuson, Jon K.; Hammel, Ken

    2011-11-01

    Abstract Brown rot basidiomycetes have an important ecological role in lignocellulose recycling and are notable for their rapid degradation of wood polymers via oxidative and hydrolytic mechanisms. However, most of these fungi apparently lack processive (exo-acting) cellulases, such as cellobiohydrolases, which are generally required for efficient cellulolysis. The recent sequencing of the Postia placenta genome now permits a proteomic approach to this longstanding conundrum. We grew P. placenta on solid aspen wood, extracted proteins from the biodegrading substrate, and analyzed tryptic digests by shotgun liquid chromatography-tandem mass spectrometry. Comparison of the data with the predicted P. placenta proteome revealed the presence of 34 likely glycoside hydrolases, but only four of these-two in glycoside hydrolase family 5, one in family 10, and one in family 12-have sequences that suggested possible activity on cellulose. We expressed these enzymes heterologously and determined that they all exhibited endoglucanase activity on phosphoric acid-swollen cellulose. They also slowly hydrolyzed filter paper, a more crystalline substrate, but the soluble/insoluble reducing sugar ratios they produced classify them as nonprocessive. Computer simulations indicated that these enzymes produced soluble/insoluble ratios on reduced phosphoric acid-swollen cellulose that were higher than expected for random hydrolysis, which suggests that they could possess limited exo activity, but they are at best 10-fold less processive than cellobiohydrolases. It appears likely that P. placenta employs a combination of oxidative mechanisms and endo-acting cellulases to degrade cellulose efficiently in the absence of a significant processive component.

  9. Genomic and experimental evidence for multiple metabolic functions in the RidA/YjgF/YER057c/UK114 (Rid) protein family

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Niehaus, Thomas D.; Gerdes, Svetlana; Hodge-Hanson, Kelsey; Zhukov, Aleksey; Cooper, Arthur J.L.; ElBadawi-Sidhu, Mona; Fiehn, Oliver; Downs, Diana M.; Hanson, Andrew D.

    2015-05-15

    It is now recognized that enzymatic or chemical side-reactions can convert normal metabolites to useless or toxic ones and that a suite of enzymes exists to mitigate such metabolite damage. Examples are the reactive imine/enamine intermediates produced by threonine dehydratase, which damage the pyridoxal 5'-phosphate cofactor of various enzymes causing inactivation. This damage is pre-empted by RidA proteins, which hydrolyze the imines before they do harm. RidA proteins belong to the YjgF/YER057c/UK114 family (here renamed the Rid family). Most other members of this diverse and ubiquitous family lack defined functions. Phylogenetic analysis divided the Rid family into a widely distributed,more » apparently archetypal RidA subfamily and seven other subfamilies (Rid1 to Rid7) that are largely confined to bacteria and often co-occur in the same organism with RidA and each other. The Rid1 to Rid3 subfamilies, but not the Rid4 to Rid7 subfamilies, have a conserved arginine residue that, in RidA proteins, is essential for imine-hydrolyzing activity. Analysis of the chromosomal context of bacterial RidA genes revealed clustering with genes for threonine dehydratase and other pyridoxal 5'-phosphate-dependent enzymes, which fits with the known RidA imine hydrolase activity. Clustering was also evident between Rid family genes and genes specifying FAD-dependent amine oxidases or enzymes of carbamoyl phosphate metabolism. Biochemical assays showed that Salmonella enterica RidA and Rid2, but not Rid7, can hydrolyze imines generated by amino acid oxidase. Genetic tests indicated that carbamoyl phosphate overproduction is toxic to S. enterica cells lacking RidA, and metabolomic profiling of Rid knockout strains showed ten-fold accumulation of the carbamoyl phosphate-related metabolite dihydroorotate. Like the archetypal RidA subfamily, the Rid2, and probably the Rid1 and Rid3 subfamilies, have imine-hydrolyzing activity and can pre-empt damage from imines formed by amine

  10. Structure of cellobiose phosphorylase from Clostridium thermocellum in complex with phosphate

    SciTech Connect (OSTI)

    Bianchetti, Christopher M.; Elsen, Nathaniel L.; Fox, Brian G.; Phillips, Jr., George N.

    2012-03-27

    Clostridium thermocellum is a cellulosome-producing bacterium that is able to efficiently degrade and utilize cellulose as a sole carbon source. Cellobiose phosphorylase (CBP) plays a critical role in cellulose degradation by catalyzing the reversible phosphate-dependent hydrolysis of cellobiose, the major product of cellulose degradation, into -D-glucose 1-phosphate and D-glucose. CBP from C. thermocellum is a modular enzyme composed of four domains [N-terminal domain, helical linker, (/)6-barrel domain and C-terminal domain] and is a member of glycoside hydrolase family 94. The 2.4 {angstrom} resolution X-ray crystal structure of C. thermocellum CBP reveals the residues involved in coordinating the catalytic phosphate as well as the residues that are likely to be involved in substrate binding and discrimination.

  11. Homologous Expression of the Caldicellulosiruptor bescii CelA Reveals that the Extracellular Protein Is Glycosylated

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Chung, Daehwan; Young, Jenna; Bomble, Yannick J.; Vander Wall, Todd A.; Groom, Joseph; Himmel, Michael E.; Westpheling, Janet

    2015-03-23

    Members of the bacterial genus Caldicellulosiruptor are the most thermophilic cellulolytic microbes described with ability to digest lignocellulosic biomass without conventional pretreatment. The cellulolytic ability of different species varies dramatically and correlates with the presence of the multimodular cellulase CelA, which contains both a glycoside hydrolase family 9 endoglucanase and a glycoside hydrolase family 48 exoglucanase known to be synergistic in their activity, connected by three cellulose-binding domains via linker peptides. This architecture exploits the cellulose surface ablation driven by its general cellulase processivity as well as excavates cavities into the surface of the substrate, revealing a novel paradigm formore » cellulase activity. We recently reported that a deletion of celA in C. bescii had a significant effect on its ability to utilize complex biomass. To analyze the structure and function of CelA and its role in biomass deconstruction, we constructed a new expression vector for C. bescii and were able, for the first time, to express significant quantities of full-length protein in vivo in the native host. The protein, which contains a Histidine tag, was active and excreted from the cell. Expression of CelA protein with and without its signal sequence allowed comparison of protein retained intracellularly to protein transported extracellularly. Analysis of protein in culture supernatants revealed that the extracellular CelA protein is glycosylated whereas the intracellular CelA is not, suggesting that either protein transport is required for this post-translational modification or that glycosylation is required for protein export. The mechanism and role of protein glycosylation in bacteria is poorly understood and the ability to express CelA in vivo in C. bescii will allow the study of the mechanism of protein glycosylation in this thermophile. Finally, it will also allow the study of glycosylation of CelA itself and its role

  12. North Shore Gas- Single Family Direct Install

    Broader source: Energy.gov [DOE]

    Owners of single-family homes, condos, townhomes and two-flats may be eligible for a free installation of new programmable thermostats, pipe insulation, showerheads, Compact Fluorescent Bulbs (CFLs...

  13. EM Helps Military Families in Need

    Office of Energy Efficiency and Renewable Energy (EERE)

    LAS VEGAS – Six families from Nellis Air Force Base in Las Vegas recently received nearly $4,000 in donations — funds that helped provide a welcome relief during the holidays.

  14. Austin Energy- Multi-Family Energy Efficiency Rebate Program

    Broader source: Energy.gov [DOE]

    The Austin Energy Multi-Family Program provides cash incentives to owners, developers, and property managers of apartments and other multi-family properties for making energy efficiency...

  15. Surface theory of a family of topological Kondo insulators (Journal...

    Office of Scientific and Technical Information (OSTI)

    Surface theory of a family of topological Kondo insulators Prev Next Title: Surface theory of a family of topological Kondo insulators Authors: Roy, Bitan ; Sau, Jay D. ; ...

  16. The Standard Work Specifications for Single-Family Home Energy...

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    The Standard Work Specifications for Single-Family Home Energy Upgrades are now available at your fingertips The Standard Work Specifications for Single-Family Home Energy ...

  17. TIGRFAMS: The TIGRFAMs database of protein families

    DOE Data Explorer [Office of Scientific and Technical Information (OSTI)]

    TIGRFAMs are protein families based on Hidden Markov Models or HMMs. Use this page to see the curated seed alignmet for each TIGRFam, the full alignment of all family members and the cutoff scores for inclusion in each of the TIGRFAMs. Also use this page to search through the TIGRFAMs and HMMs for text in the TIGRFAMs Text Search or search for specific sequences in the TIGRFAMs Sequence Search.[Copied from the Overview at http://www.jcvi.org/cms/research/projects/tigrfams/overview/] See also TIGRFAMs ordered by the roles they play at http://cmr.jcvi.org/tigr-scripts/CMR/shared/EvidenceList.cgi?ev_type=TIGRFAM&order_type=role.

  18. Genetic linkage analysis in familial breast and ovarian cancer: Results from 214 families

    SciTech Connect (OSTI)

    Easton, D.F.; Ford, D. ); Bishop, D.T.; Crockford, G.P. )

    1993-04-01

    This paper reports the results of a collaborative linkage study involving 214 breast cancer families, including 57 breast-ovarian cancer families; this represents almost all the known families with 17q linkage data. Six markers on 17q, spanning approximately 30 cM, were typed in the families. The aims of the study were to define more precisely the localization of the disease gene, the extent of genetic heterogeneity and the characteristics of linked families and to estimate the penetrance of the 17q gene. Under the assumption of no genetic heterogeneity, the strongest linkage evidence was obtained with D17S588. Multipoint linkage analysis allowing for genetic heterogeneity provided evidence that the predisposing gene lies between the markers D17S588 and D17S250, an interval whose genetic length is estimated to be 8.3 cM in males and 18.0 cM in females. This position was supported over other intervals by odds of 66:1. The location of the gene with respect to D17S579 could not be determined unequivocally. Under the genetic model used in the analysis, the best estimate of the proportion of linked breast-ovarian cancer families was 1.0 (lower LOD -- 1 limit 0.79). In contrast, there was significant evidence of genetic heterogeneity among the families without ovarian cancer, with an estimated 45% being linked. These results suggest that a gene(s) on chromosome 17q accounts for the majority of families in which both early-onset breast cancer and ovarian cancer occur but that other genes predisposing to breast cancer exist. By examining the fit of the linkage data to different penetrance functions, the cumulative risk associated with the 17q gene was estimated to be 59% by age 50 years and 82% by age 70 years. The corresponding estimates for the breast-ovary families were 67% and 76%, and those for the families without ovarian cancer were 49% and 90%; these penetrance functions did not differ significantly from one another. 42 refs., 5 figs., 2 tabs.

  19. Better Buildings Neighborhood Program Multi-Family Peer Exchange...

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Neighborhood Program Multi- Family Peer Exchange Call: Shared Space vs. In- unit Upgrades ... focusing on shared space and technology in multi-family buildings vs. in-unit upgrades? ...

  20. Be SMART Multi-Family Efficiency Loan Program

    Broader source: Energy.gov [DOE]

    Under the Be SMART Multi-Family Program, the Maryland Department of Housing and Community Development (DHCD) offers loans for energy efficiency improvements in existing multi-family rental proper...

  1. Application for State Land Use Lease: Commercial/Multi-Family...

    Open Energy Info (EERE)

    CommercialMulti-Family Jump to: navigation, search OpenEI Reference LibraryAdd to library Reference: Application for State Land Use Lease: CommercialMulti-Family Published...

  2. Atomic veterans and their families: Responses to radiation exposure

    SciTech Connect (OSTI)

    Murphy, B.C.; Ellis, P.; Greenberg, S. )

    1990-07-01

    In-depth interviews with seven atomic veterans and their families indicated powerful psychological effects on all family members from exposure to low-level ionizing radiation. Four themes emerged: the invalidation of their experiences by government and other authority figures; family concerns about genetic effects on future generations; family members' desire to protect each other from fears of physical consequences; and desire to leave a record of their experiences to help prevent future suffering.

  3. Vladimir V. Lunin | Bioenergy | NREL

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Vladimir V. Lunin Vladimir V. Lunin Senior Scientist, Protein Crystallography Vladimir.Lunin@nrel.gov | 303-384-7754 Research Interests Glycoside hydrolases, lyases, transferases Establishing molecular mechanisms of protein-substrate interaction for enzymes involved in polysaccharide synthesis, modification, or breakdown Biomass deconstruction and recalcitrance Areas of Expertise Protein purification Protein crystallization X-ray diffraction data collection Protein structure determination

  4. Page 7, A Family-Friendly Workplace

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    7 of 11 Previous Page A Family-Friendly Workplace The Department of Energy (DOE) takes its commitment to recruiting, retaining, and motivating a highly skilled, diverse, and productive workforce very seriously. Our objective is to be the employer of choice in the public sector. The U.S. Department of Energy is the Top-Ranked Cabinet-Level Agency for employee satisfaction and engagement according to the 2005 Best Places to Work in the Federal Government Rankings. Below is a summary of additional

  5. Leptonic mixing, family symmetries, and neutrino phenomenology

    SciTech Connect (OSTI)

    Medeiros Varzielas, I. de [Departamento de Fisica and Centro de Fisica Teorica de Particulas, Instituto Superior Tecnico, Avenida Rovisco Pais, 1049-001 Lisboa (Portugal); Fakultaet fuer Physik, Technische Universitaet Dortmund D-44221 Dortmund (Germany); Gonzalez Felipe, R. [Departamento de Fisica and Centro de Fisica Teorica de Particulas, Instituto Superior Tecnico, Avenida Rovisco Pais, 1049-001 Lisboa (Portugal); Instituto Superior de Engenharia de Lisboa, Rua Conselheiro Emidio Navarro, 1959-007 Lisboa (Portugal); Serodio, H. [Departamento de Fisica and Centro de Fisica Teorica de Particulas, Instituto Superior Tecnico, Avenida Rovisco Pais, 1049-001 Lisboa (Portugal)

    2011-02-01

    Tribimaximal leptonic mixing is a mass-independent mixing scheme consistent with the present solar and atmospheric neutrino data. By conveniently decomposing the effective neutrino mass matrix associated to it, we derive generic predictions in terms of the parameters governing the neutrino masses. We extend this phenomenological analysis to other mass-independent mixing schemes which are related to the tribimaximal form by a unitary transformation. We classify models that produce tribimaximal leptonic mixing through the group structure of their family symmetries in order to point out that there is often a direct connection between the group structure and the phenomenological analysis. The type of seesaw mechanism responsible for neutrino masses plays a role here, as it restricts the choices of family representations and affects the viability of leptogenesis. We also present a recipe to generalize a given tribimaximal model to an associated model with a different mass-independent mixing scheme, which preserves the connection between the group structure and phenomenology as in the original model. This procedure is explicitly illustrated by constructing toy models with the transpose tribimaximal, bimaximal, golden ratio, and hexagonal leptonic mixing patterns.

  6. Saving American Families and Businesses Money through Lighting Efficiency |

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Department of Energy Saving American Families and Businesses Money through Lighting Efficiency Saving American Families and Businesses Money through Lighting Efficiency February 12, 2016 - 1:38pm Addthis Saving American Families and Businesses Money through Lighting Efficiency David Friedman David Friedman Assistant Secretary for Energy Efficiency and Renewable Energy (Acting) What are the key facts? The Department of Energy released a new proposed efficiency rule for lightbulbs to make it

  7. Rural Development Multi-Family Housing Energy Efficiency Initiative |

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Department of Energy Rural Development Multi-Family Housing Energy Efficiency Initiative Rural Development Multi-Family Housing Energy Efficiency Initiative In order to help create a more energy independent rural America for the next century, the USDA Rural Development Multi-Family Housing Energy Efficiency Initiative enables applicants to several USDA housing programs to increase their program funding eligibility by incorporating green building practices into project designs, construction,

  8. Alabama Family Staying Nice and Cozy This Fall

    Broader source: Energy.gov [DOE]

    Recovery Act money to weatherize homes has resulted in much lower energy bills for Alabama families, including Mary, whose bill is about $300 cheaper now.

  9. A family of GE engineers | GE Global Research

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    A family of GE engineers Click to email this to a friend (Opens in new window) Share on Facebook (Opens in new window) Click to share (Opens in new window) Click to share on LinkedIn (Opens in new window) Click to share on Tumblr (Opens in new window) A family of GE engineers Father's Day is a time dedicated to celebrating family ties. In the GE family, there are many types of relationships and connections, including some that originated in the same household. Meet Monte and children, Ashlee and

  10. NNSA Production Office tops Feds Feed Families campaign goal...

    National Nuclear Security Administration (NNSA)

    tops Feds Feed Families campaign goal | National Nuclear Security Administration Facebook Twitter Youtube Flickr RSS People Mission Managing the Stockpile Preventing Proliferation...

  11. Saving American Families and Businesses Money through Lighting...

    Broader source: Energy.gov (indexed) [DOE]

    Saving American Families and Businesses Money through Lighting Efficiency David Friedman ... to make cost-effective, energy efficient choices for lighting their homes and offices. ...

  12. Willi Ernst and family members | Open Energy Information

    Open Energy Info (EERE)

    founder and Managing Director. He sold his shares to Centrosolar but continues to run Biohaus. References: Willi Ernst (and family members)1 This article is a stub. You...

  13. Structures of Cyanobactin Maturation Enzymes Define a Family...

    Office of Scientific and Technical Information (OSTI)

    Structures of Cyanobactin Maturation Enzymes Define a Family of Transamidating Proteases Citation Details In-Document Search Title: Structures of Cyanobactin Maturation Enzymes ...

  14. Energy Auditor - Single Family 2.0: General Construction and...

    Energy Savers [EERE]

    General Construction and Carpentry Energy Auditor - Single Family 2.0: General Construction and Carpentry General Construction and Carpentry - Complete (3.02 MB) Lesson Plan: ...

  15. Synthesis, Electrochemistry and Photophysics of a Family of Phlorin...

    Office of Scientific and Technical Information (OSTI)

    Journal Article: Synthesis, Electrochemistry and Photophysics of a Family of Phlorin Macrocycles that Display Cooperative Fluoride Binding Citation Details In-Document Search ...

  16. Better Buildings Neighborhood Program Multi-family/ Low Income...

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    ... 3152012 3152012 3152012 3152012 GIS Mapping pre-1990 construction 3152012 3... The GIS department could then cross- reference properties that are coded as multi-family ...

  17. The Energy Department Feeds Families | Department of Energy

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    ... Last year, Energy Department employee Tyesha Bennett and her family recently gleaned 4,400 pounds of sweet potatoes from Cheltenham Warehouse in Maryland -- meaning more fresh, ...

  18. SoCalGas- Multi-Family Residential Rebate Program

    Broader source: Energy.gov [DOE]

    Southern California Gas Company provides incentives to encourage the owners and managers of multi-family residential buildings to increase their energy efficiency. The program offers rebates for...

  19. Standard Work Specifications for Single-Family Home Energy Upgrades...

    Energy Savers [EERE]

    stakeholders developed the Standard Work Specifications for Single-Family Home Energy Upgrades to define the minimum requirements for high-quality residential energy upgrades. ...

  20. BRCA1-linked marker in postmenopausal breast cancer families

    SciTech Connect (OSTI)

    Folsom, A.R.; Chen, P.L.; Sellers, T.A.

    1994-09-01

    A majority of breast and ovarian cancer families and half of the early-onset breast cancer families are linked to markers on 17q (BRCA1). While linkage has been demonstrated in families with premenopausal disease, few studies have tested these markers in families with postmenopausal breast cancer. In the Iowa Women`s Health Study, a population-based study of over 42,000 women, an association of waist-to-hip ratio (WHR) with the risk of postmenopausal breast cancer was found predominantly in women with a positive family history -- this interaction was associated with a 3.2-fold elevated risk. This effect was even more pronounced when the definition of family history included breast and ovarian cancer, known to be linked to 17q markers. We evaluated evidence for linkage with D17S579, a BRCA-1-linked marker, in 13 families in which the index case had postmenopausal breast cancer. Genotyping for alleles at D17S579 was performed on 84 blood samples. Linkage analysis assumed that the breast cancer trait had an autosomal dominant mode of inheritance with a penetrance of 80%. For the 13 families studied, the maximum lod score was 0.29 at a theta of 0.27. There was significant evidence against tight linkage of breast cancer with D17S579 (theta<0.4). Heterogeneity analysis suggested evidence for the presence of both linked and unlinked families. Partitioning informative families on WHR of the index case suggested heterogeneity. These data suggest that, in a subset of families identified by a postmenopausal breast cancer proband, risk of breast cancer may be mediated by BRCA1, with heterogeneity defined by WHR.

  1. Linkage heterogeneity among 59 Dutch hereditary breast cancer families

    SciTech Connect (OSTI)

    Cornelis, R.S.; Vliet, M. van; Leeuwen, I. van

    1994-09-01

    We have investigated 59 Dutch kindreds with at least three first-degree relatives with breast and/or ovarian cancer for linkage to BRCA1 on 17q12-q21, using at least 4 microsatellite markers flanking BRCA1 on either side. Assuming no heterogeneity, the overall multipoint lod score in this group of families was -7.59. A marked clustering of lod scores >0.5 was observed among the 13 families with a mean age of onset lower than 45 (total lod score: +3.36). Among the 8 kindreds with a mean age of onset lower than 45 and {ge}3 cases diagnosed under 45, the lod score was +4.43. Interestingly, most of the evidence against linkage was found in 17 families with a mean age of onset between 45 and 54 (total lod score of -8.72). It was estimated that 28% of the breast-only families might be caused by BRCA1. Over the 16 breast-ovarian cancer families a lod score of -3.78 was obtained under homogeneity. The highest lod score was +0.57, assuming heterogeneity with 33% of the families being linked to BRCA1. One family gave a multipoint lod score of -2.01 and thereby satisfies the conventional criterion of an unlinked family. Our results support the conclusions from earlier work by others, namely that BRCA1 predisposes particularly to early-onset breast cancer. The proportion of breast-ovarian cancer families we found linked to BRCA1 is much lower than that found by the Breast Cancer Linkage Consortium. This might be caused by the single unlinked family against an insufficient number of families able to give conclusive positive lod scores.

  2. Caldicellulosiruptor Core and Pangenomes Reveal Determinants for

    SciTech Connect (OSTI)

    Blumer-Schuette, Sara E.; Giannone, Richard J; Zurawski, Jeffrey V; Ozdemir, Inci; Ma, Qin; Yin, Yanbin; Xu, Ying; Kataeva, Irena; Poole, Farris; Adams, Michael W. W.; Hamilton-Brehm, Scott; Elkins, James G; Larimer, Frank W; Land, Miriam L; Hauser, Loren John; Cottingham, Robert W; Hettich, Robert {Bob} L; Kelly, Robert M

    2012-01-01

    Extremely thermophilic bacteria of the genus Caldicellulosiruptor utilize carbohydrate components of plant cell walls, including cellulose and hemicellulose, facilitated by a diverse set of glycoside hydrolases (GHs). From a biofuel perspective, this capability is crucial for deconstruction of plant biomass into fermentable sugars. While all species from the genus grow on xylan and acidpretreated switchgrass, growth on crystalline cellulose is variable. The basis for this variability was examined using microbiological, genomic, and proteomic analyses of eight globally diverse Caldicellulosiruptor species. The open Caldicellulosiruptor pangenome (4,009 open reading frames [ORFs]) encodes 106 GHs, representing 43 GH families, but only 26 GHs from 17 families are included in the core (noncellulosic) genome (1,543 ORFs). Differentiating the strongly cellulolytic Caldicellulosiruptor species from the others is a specific genomic locus that encodes multidomain cellulases from GH families 9 and 48, which are associated with cellulose-binding modules. This locus also encodes a novel adhesin associated with type IV pili, which was identified in the exoproteome bound to crystalline cellulose. Taking into account the core genomes, pangenomes, and individual genomes, the ancestral Caldicellulosiruptor was likely cellulolytic and evolved, in some cases, into species that lost the ability to degrade crystalline cellulose while maintaining the capacity to hydrolyze amorphous cellulose and hemicellulose.

  3. Jefferson Lab Invites Families, Groups, Classes to Physics Fest Events |

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Jefferson Lab Families, Groups, Classes to Physics Fest Events Jefferson Lab Invites Families, Groups, Classes to Physics Fest Events Small Business A student watches in amazement at the behavior of inflated balloons after they have been pulled from a container of liquid nitrogen (at a temperature of -321 Fahrenheit). NEWPORT NEWS, Va., June 3, 2010 - The U.S. Department of Energy's Jefferson Lab invites families, groups and classes to sign up now for a summer 2010 or school year 2010-2011

  4. Suite of Activity-Based Probes for Cellulose-Degrading Enzymes

    SciTech Connect (OSTI)

    Chauvigne-Hines, Lacie M.; Anderson, Lindsey N.; Weaver, Holly M.; Brown, Joseph N.; Koech, Phillip K.; Nicora, Carrie D.; Hofstad, Beth A.; Smith, Richard D.; Wilkins, Michael J.; Callister, Stephen J.; Wright, Aaron T.

    2012-12-19

    Microbial glycoside hydrolases play a dominant role in the biochemical conversion of cellulosic biomass to high-value biofuels. Anaerobic cellulolytic bacteria are capable of producing multicomplex catalytic subunits containing cell-adherent cellulases, hemicellulases, xylanases, and other glycoside hydrolases to facilitate the degradation of highly recalcitrant cellulose and other related plant cell wall polysaccharides. Clostridium thermocellum is a cellulosome producing bacterium that couples rapid reproduction rates to highly efficient degradation of crystalline cellulose. Herein, we have developed and applied a suite of difluoromethylphenyl aglycone, N-halogenated glycosylamine, and 2-deoxy-2-fluoroglycoside activity-based protein profiling (ABPP) probes to the direct labeling of the C. thermocellum cellulosomal secretome. These activity-based probes (ABPs) were synthesized with alkynes to harness the utility and multimodal possibilities of click chemistry, and to increase enzyme active site inclusion for LC-MS analysis. We directly analyzed ABP-labeled and unlabeled global MS data, revealing ABP selectivity for glycoside hydrolase (GH) enzymes in addition to a large collection of integral cellulosome-containing proteins. By identifying reactivity and selectivity profiles for each ABP, we demonstrate our ability to widely profile the functional cellulose degrading machinery of the bacterium. Derivatization of the ABPs, including reactive groups, acetylation of the glycoside binding groups, and mono- and disaccharide binding groups, resulted in considerable variability in protein labeling. Our probe suite is applicable to aerobic and anaerobic cellulose degrading systems, and facilitates a greater understanding of the organismal role associated within biofuel development.

  5. A Nuclear Family: Extras - Introduction | Y-12 National Security...

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    ... A Nuclear Family: Extras - Introduction The mp4 video format is not supported by this browser. Download video Captions: On Time: 6:59 min. D. Ray Smith, Y-12 historian,...

  6. Sam's Family Spa Pool & Spa Low Temperature Geothermal Facility...

    Open Energy Info (EERE)

    of Technology's Geo-Heat Center Retrieved from "http:en.openei.orgwindex.php?titleSam%27sFamilySpaPool%26SpaLowTemperatureGeothermalFacility&oldid305482" ...

  7. Secretaries Chu and Donovan Sign Agreement to Help Working Families

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Weatherize Their Homes | Department of Energy Donovan Sign Agreement to Help Working Families Weatherize Their Homes Secretaries Chu and Donovan Sign Agreement to Help Working Families Weatherize Their Homes May 6, 2009 - 12:00am Addthis WASHINGTON - U.S. Department of Housing and Urban Development (HUD) Secretary Shaun Donovan and U.S. Department of Energy (DOE) Secretary Steven Chu today announced an important step forward in the partnership between HUD and DOE to streamline and better

  8. Secretaries Donovan and Chu Announce Partnership to Help Working Families

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Weatherize Their Homes | Department of Energy Donovan and Chu Announce Partnership to Help Working Families Weatherize Their Homes Secretaries Donovan and Chu Announce Partnership to Help Working Families Weatherize Their Homes February 27, 2009 - 12:00am Addthis Philadelphia, PA - U.S. Department of Housing and Urban Development (HUD) Secretary Shaun Donovan and U.S. Department of Energy (DOE) Secretary Steven Chu today announced a major partnership between HUD and DOE that will streamline

  9. Weatherization Saves Families Energy and Money | Department of Energy

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Weatherization Saves Families Energy and Money Weatherization Saves Families Energy and Money October 28, 2013 - 12:50pm Addthis William Stewart, with Veterans Green Jobs, blows cellulose insulation in the interior walls of a Lakewood, Colorado, home. This home is part of the Energy Department's Weatherization Assistance Program that supports energy efficiency upgrades to low-income homes in Denver. | Photo courtesy of Dennis Schroeder, NREL. William Stewart, with Veterans Green Jobs, blows

  10. Utilities Bringing More Solar Energy to Families and Businesses |

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Department of Energy Utilities Bringing More Solar Energy to Families and Businesses Utilities Bringing More Solar Energy to Families and Businesses January 26, 2016 - 12:26pm Addthis This two megawatt installation at Denver International Airport is owned by Constellation Energy, a utility based in Baltimore, MD. (Photo Courtesy: Denver International Airport) This two megawatt installation at Denver International Airport is owned by Constellation Energy, a utility based in Baltimore, MD.

  11. A Family-Friendly Workplace | Department of Energy

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Services » New Employee Orientation » A Family-Friendly Workplace A Family-Friendly Workplace The Department of Energy (DOE) takes its commitment to recruiting, retaining, and motivating a highly skilled, diverse, and productive workforce very seriously. Our objective is to be the employer of choice in the public sector. The U.S. Department of Energy is the Top-Ranked Cabinet-Level Agency for employee satisfaction and engagement according to the 2005 Best Places to Work in the Federal

  12. Functional and structural diversity in GH62 α-L-arabinofuranosidases from the thermophilic fungus Scytalidium thermophilum

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Kaur, Amrit Pal; Nocek, Boguslaw P.; Xu, Xiaohui; Lowden, Michael J.; Leyva, Juan Francisco; Stogios, Peter J.; Cui, Hong; Leo, Rosa Di; Powlowski, Justin; Tsang, Adrian; et al

    2015-05-01

    The genome of the thermophilic fungus Scytalidium thermophilum (strain CBS 625.91) harbours a wide range of genes involved in carbohydrate degradation, including three genes, abf62A, abf62B and abf62C, predicted to encode glycoside hydrolase family 62 (GH62) enzymes. Transcriptome analysis showed that only abf62A and abf62C are actively expressed during growth on diverse substrates including straws from barley, alfalfa, triticale and canola. The abf62A and abf62C genes were expressed in Escherichia coli and the resulting recombinant proteins were characterized. Calcium-free crystal structures of Abf62C in apo and xylotriose bound forms were determined to 1.23 and 1.48 Å resolution respectively. Site-directed mutagenesismore » confirmed Asp55, Asp171 and Glu230 as catalytic triad residues, and revealed the critical role of non-catalytic residues Asp194, Trp229 and Tyr338 in positioning the scissile α-L-arabinofuranoside bond at the catalytic site. Further, the +2R substrate-binding site residues Tyr168 and Asn339, as well as the +2NR residue Tyr226, are involved in accommodating long-chain xylan polymers. Overall, our structural and functional analysis highlights characteristic differences between Abf62A and Abf62C, which represent divergent subgroups in the GH62 family.« less

  13. Conversion of levoglucosan and cellobiosan by Pseudomonas putida KT2440

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Linger, Jeffrey G.; Hobdey, Sarah E.; Franden, Mary Ann; Fulk, Emily M.; Beckham, Gregg T.

    2016-02-02

    Pyrolysis offers a straightforward approach for the deconstruction of plant cell wall polymers into bio-oil. Recently, there has been substantial interest in bio-oil fractionation and subsequent use of biological approaches to selectively upgrade some of the resulting fractions. A fraction of particular interest for biological upgrading consists of polysaccharide-derived substrates including sugars and sugar dehydration products such as levoglucosan and cellobiosan, which are two of the most abundant pyrolysis products of cellulose. Levoglucosan can be converted to glucose-6-phosphate through the use of a levoglucosan kinase (LGK), but to date, the mechanism for cellobiosan utilization has not been demonstrated. Here, wemore » engineer the microbe Pseudomonas putida KT2440 to use levoglucosan as a sole carbon and energy source through LGK integration. Furthermore, we demonstrate that cellobiosan can be enzymatically converted to levoglucosan and glucose with β-glucosidase enzymes from both Glycoside Hydrolase Family 1 and Family 3. β-glucosidases are commonly used in both natural and industrial cellulase cocktails to convert cellobiose to glucose to relieve cellulase product inhibition and to facilitate microbial uptake of glucose. Using an exogenous β-glucosidase, we demonstrate that the engineered strain of P. putida can grow on levoglucosan up to 60 g/L and can also utilize cellobiosan. Overall, this study elucidates the biological pathway to co-utilize levoglucosan and cellobiosan, which will be a key transformation for the biological upgrading of pyrolysis-derived substrates.« less

  14. Purification, crystallization and crystallographic analysis of Clostridium thermocellum endo-1,4-β-d-xylanase 10B in complex with xylohexaose

    SciTech Connect (OSTI)

    Najmudin, Shabir; Pinheiro, Benedita A.; Romão, Maria J.; Prates, José A. M.; Fontes, Carlos M. G. A.

    2008-08-01

    The N-terminal moiety of C. thermocellum endo-1,4-β-d-xylanase 10B, comprising a carbohydrate-binding module (CBM22-1) and a GH10 E337A mutant domain, has been crystallized in complex with xylohexaose. The crystals belong to the trigonal space group P3{sub 2}21, contain a dimer in the asymmetric unit and diffract to beyond 2.0 Å resolution. The cellulosome of Clostridium thermocellum is a highly organized multi-enzyme complex of cellulases and hemicellulases involved in the hydrolysis of plant cell-wall polysaccharides. The bifunctional multi-modular xylanase Xyn10B is one of the hemicellulase components of the C. thermocellum cellulosome. The enzyme contains an internal glycoside hydrolase family 10 catalytic domain (GH10) and a C-terminal family 1 carbohydrate esterase domain (CE1). The N-terminal moiety of Xyn10B (residues 32–551), comprising a carbohydrate-binding module (CBM22-1) and the GH10 E337A mutant, was crystallized in complex with xylohexaose. The crystals belong to the trigonal space group P3{sub 2}21 and contain a dimer in the asymmetric unit. The crystals diffracted to beyond 2.0 Å resolution.

  15. Detection of eight BRCA1 mutations in 10 breast/ovarian cancer families, including 1 family with male breast cancer

    SciTech Connect (OSTI)

    Sruewing, J.P.; Brody, L.C.; Erdos, M.R.

    1995-07-01

    Genetic epidemiological evidence suggests that mutations in BRCA1 may be responsible for approximately one half of early onset familial breast cancer and the majority of familial breast/ovarian cancer. The recent cloning of BRCA1 allows for the direct detection of mutations, but the feasibility of presymptomatic screening for cancer susceptibility is unknown. We analyzed genomic DNA from one affected individual from each of 24 families with at least three cases of ovarian or breast cancer, using SSCP assays. Variant SSCP bands were subcloned and sequenced. Allele-specific oligonucleotide hybridization was used to verify sequence changes and to screen DNA from control individuals. Six frameshift and two missense mutations were detected in 10 different families. A frameshift mutation was detected in a male proband affected with both breast and prostate cancer. A 40-bp deletion was detected in a patient who developed intra-abdominal carcinomatosis 1 year after prophylactic oophorectomy. Mutations were detected throughout the gene, and only one was detected in more than a single family. These results provide further evidence that inherited breast and ovarian cancer can occur as a consequence of a wide array of BRCA1 mutations. These results suggests that development of a screening test for BRCA1 mutations will be technically challenging. The finding of a mutation in a family with male breast cancer, not previously thought to be related to BRCA1, also illustrates the potential difficulties of genetic counseling for individuals known to carry mutations. 37 refs., 1 fig., 1 tab.

  16. Evolutionary analyses of non-family genes in plants

    SciTech Connect (OSTI)

    Ye, Chuyu; Li, Ting; Yin, Hengfu; Weston, David; Tuskan, Gerald A; Tschaplinski, Timothy J; Yang, Xiaohan

    2013-03-01

    There are a large number of non-family (NF) genes that do not cluster into families with three or more members per genome. While gene families have been extensively studied, a systematic analysis of NF genes has not been reported. We performed comparative studies on NF genes in 14 plant species. Based on the clustering of protein sequences, we identified ~94,000 NF genes across these species that were divided into five evolutionary groups: Viridiplantae-wide, angiosperm-specific, monocot-specific, dicot-specific, and those that were species-specific. Our analysis revealed that the NF genes resulted largely from less frequent gene duplications and/or a higher rate of gene loss after segmental duplication relative to genes in both low-copy-number families (LF; 3 10 copies per genome) and high-copy-number families (HF; >10 copies). Furthermore, we identified functions enriched in the NF gene set as compared with the HF genes. We found that NF genes were involved in essential biological processes shared by all plant lineages (e.g., photosynthesis and translation), as well as gene regulation and stress responses associated with phylogenetic diversification. In particular, our analysis of an Arabidopsis protein-protein interaction network revealed that hub proteins with the top 10% most connections were over-represented in the NF set relative to the HF set. This research highlights the roles that NF genes may play in evolutionary and functional genomics research.

  17. Evolutionary analyses of non-family genes in plants

    SciTech Connect (OSTI)

    Ye, Chuyu; Li, Ting; Yin, Hengfu; Weston, David; Tuskan, Gerald A; Tschaplinski, Timothy J; Yang, Xiaohan

    2013-01-01

    There are a large number of non-family (NF) genes that do not cluster into families with three or more members per genome. While gene families have been extensively studied, a systematic analysis of NF genes has not been reported. We performed comparative studies on NF genes in 14 plant species. Based on the clustering of protein sequences, we identified ~94 000 NF genes across these species that were divided into five evolutionary groups: Viridiplantae wide, angiosperm specific, monocot specific, dicot specific, and those that were species specific. Our analysis revealed that the NF genes resulted largely from less frequent gene duplications and/or a higher rate of gene loss after segmental duplication relative to genes in both lowcopy- number families (LF; 3 10 copies per genome) and high-copy-number families (HF; >10 copies). Furthermore, we identified functions enriched in the NF gene set as compared with the HF genes. We found that NF genes were involved in essential biological processes shared by all plant lineages (e.g. photosynthesis and translation), as well as gene regulation and stress responses associated with phylogenetic diversification. In particular, our analysis of an Arabidopsis protein protein interaction network revealed that hub proteins with the top 10% most connections were over-represented in the NF set relative to the HF set. This research highlights the roles that NF genes may play in evolutionary and functional genomics research.

  18. A Common Catalytic Mechanism for Proteins of the HutI Family

    SciTech Connect (OSTI)

    Tyagi,R.; Eswaramoorthy, S.; Burley, S.; Raushel, F.; Swaminathan, S.

    2008-01-01

    Imidazolonepropionase (HutI) (imidazolone-5-propanote hydrolase, EC 3.5.2.7) is a member of the amidohydrolase superfamily and catalyzes the conversion of imidazolone-5-propanoate to N-formimino-l-glutamate in the histidine degradation pathway. We have determined the three-dimensional crystal structures of HutI from Agrobacterium tumefaciens (At-HutI) and an environmental sample from the Sargasso Sea Ocean Going Survey (Es-HutI) bound to the product [N-formimino-l-glutamate (NIG)] and an inhibitor [3-(2, 5-dioxoimidazolidin-4-yl)propionic acid (DIP)], respectively. In both structures, the active site is contained within each monomer, and its organization displays the landmark feature of the amidohydrolase superfamily, showing a metal ligand (iron), four histidines, and one aspartic acid. A catalytic mechanism involving His265 is proposed on the basis of the inhibitor-bound structure. This mechanism is applicable to all HutI forms.

  19. High quality draft genome sequence of Flavobacterium rivuli type strain WB 3.3-2T (DSM 21788T), a valuable source of polysaccharide decomposing enzymes

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Hahnke, Richard L.; Stackebrandt, Erko; Meier-Kolthoff, Jan P.; Tindall, Brian J.; Huang, Sixing; Rohde, Manfred; Lapidus, Alla; Han, James; Trong, Stephan; Haynes, Matthew; et al

    2015-07-30

    Flavobacterium rivuli Ali et al. 2009 emend. Dong et al. 2013 is one of about 100 species in the genus Flavobacterium (family Flavobacteriacae, phylum Bacteroidetes) with a validly published name, and has been isolated from the spring of a hard water rivulet in Northern Germany. Including all type strains of the genus Myroides and Flavobacterium into the 16S rRNA gene sequence phylogeny revealed a clustering of members of the genus Myroides as a monophyletic group within the genus Flavobacterium. Furthermore, F. rivuli WB 3.3-2T and its next relatives seem more closely related to the genus Myroides than to the typemore » species of the genus Flavobacterium, F. aquatile. The 4,489,248 bp long genome with its 3,391 protein-coding and 65 RNA genes is part of the G enomic E ncyclopedia of B acteria and A rchaea project. The genome of F. rivuli has almost as many genes encoding carbohydrate active enzymes (151 CAZymes) as genes encoding peptidases (177). Peptidases comprised mostly metallo (M) and serine (S) peptidases. Among CAZymes, 30 glycoside hydrolase families, 10 glycosyl transferase families, 7 carbohydrate binding module families and 7 carbohydrate esterase families were identified. Furthermore, we found four polysaccharide utilization loci (PUL) and one large CAZy rich gene cluster that might enable strain WB 3.3-2T to decompose plant and algae derived polysaccharides. In conclusion, based on these results we propose F. rivuli as an interesting candidate for further physiological studies and the role of Bacteroidetes in the decomposition of complex polymers in the environment.« less

  20. Family Moderate Income Homeowners In New York State

    Broader source: Energy.gov [DOE]

    "Family Moderate Income Homeowners In New York State: Enhancing Resource Accessibility Through Process Improvement and Targeted Outreach," by Residential Energy Efficiency Solutions, July 10, 2012, Arlington, Virginia. Provides an overview of broadening accessibility to financing through process improvement and targeted outreach.

  1. HUD rehabilitation energy guidelines for multi-family dwellings

    SciTech Connect (OSTI)

    1996-09-01

    The Guidebook has been prepared to help people, like yourself, involved in the rehabilitation of a multi-family building. It will help you understand the basics of residential energy conservation, and to think positively about the potential economic benefits. The Guidebook, and the HUD Rehabilitation Energy Guidelines, are applicable to all climates from hot and humid Florida to the northern reaches of Alaska.

  2. Experimental plan for the Single-Family Study

    SciTech Connect (OSTI)

    Berry, L.G.; Brown, M.A.; Wright, T.; White, D.L.

    1991-09-01

    The national evaluation of the Weatherization Assistance Program (WAP) consists of five separate studies. The Single-Family Study is one of three studies that will estimate program energy savings and cost effectiveness in principal WAP submarkets. This report presents the experimental plan for the Single-Family Study, which will be implemented over the next three years (1991--1993). The Single-Family Study will directly estimate energy savings for a nationally representative sample of single-family and small multifamily homes weatherized in the 1989 program year. Savings will be estimated from gas and electric utility billing records using the Princeton Scorekeeping Method (PRISM). The study will also assess nonenergy impacts (e.g., health, comfort, safety, and housing affordability), estimate cost effectiveness, and analyze factors influencing these outcomes. For homes using fuels such as wood, coal, fuel oil, kerosene, and propane as the primary source of space conditioning, energy savings will be studied indirectly. The study will assemble a large nationally representative data base. A cluster sampling approach will be used, in which about 400 subgrantees are selected in a first stage and weatherized homes are selected in a second range. To ensure that the Single-Family Study is able to identify promising opportunities for future program development, two purposively selected groups of subgrantees will be included: (1) subgrantees that install cooling measures (such as more efficient air conditioning equipment or radiant barriers), and (2) exemplary subgrantees that use state-of-the-art technologies and service delivery procedures (such as advanced audit techniques, blower door tests, infrared scanners, extensive client education, etc.). These two groups of subgrantees will be analyzed to identify the most effective program elements in specific circumstances. 14 refs., 4 figs., 3 tabs.

  3. Hui Wei | Bioenergy | NREL

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Hui Wei Hui Wei Research Scientist Hui.Wei@nrel.gov | 303-384-6620 Research Interests Genetic modification of plants with glycoside hydrolase and biocatalyst overexpression to increase the biomass pretreatability and digestibility Genetic engineering of yeast and bacteria for the production of advanced biofuels Biomaterials and biomanufacturing Affiliated Research Programs Targeted Microbial Development Advanced concepts for producing hydrocarbons, 2015-present Targeted Conversion Research

  4. Brandon Knott | Bioenergy | NREL

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Brandon Knott Brandon Knott Staff Engineer Brandon.Knott@nrel.gov | 303-384-6223 Research Interests Molecular mechanisms of cellulose biosynthesis Molecular mechanisms of cellulose-degrading enzymes Structure-function relationships in glycoside hydrolases Techno-economic analysis (TEA) of biomass conversion technologies Methane clathrate hydrates formation Molecular Mechanisms of Cellulose Biosynthesis Illustration of the domains of the synthase enzyme, represented as Cellulose is the primary

  5. Living in the Nuclear Age: Families and the threat of nuclear war

    SciTech Connect (OSTI)

    Demorest, D.M.

    1991-01-01

    The main interest of this research was to add to the body of knowledge about the possible psychological impact of the nuclear threat on the family unit. Data were utilized from the Family Interaction, Stress and Nuclear War study conducted by Jules Riskin, M.D. and Victoria Dickerson, Ph.D. at the Mental Research Institute in Palo Alto, California. The sample consisted of ten families who were recruited for this study. In order to examine family-interaction variables and the impact of the threat of nuclear war, a standardized semi-structured family interview was conducted. Topics ranged from ordinary activities to external, non-nuclear stresses such as landslides or hurricanes, to the topic of nuclear war. A distinction is drawn between a family's level of nuclear concern while they discuss nuclear issues and a family's level of nuclear concern when viewed in the context of their overall pattern of family communication. In terms of family coping, family nuclear concern was found to be significantly related to two family-coping strategies. Families who utilized the coping strategies of seeking spiritual support and mobilizing the family to acquire and accept help were significantly less concerned about the threat of nuclear war.

  6. Lynden Archer > James A. Friend Family Distinguished Professor

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Chemical and Biomolecular Engineering > Faculty Directory > The Energy Materials Center at Cornell Lynden Archer James A. Friend Family Distinguished Professor Chemical and Biomolecular Engineering Research Group Webpage laa25@cornell.edu Professor Archer received the Career Award from the National Science Foundation, 1996, Dupont Young Professor Award 1996-1999, 3M Company Non-Tenured Faculty Award 1995, and the George Armistead Faculty Fellowship 1999-2000. Lynden joined the Cornell

  7. Jefferson Lab invites families, groups to Summer 2005 Physics Fests |

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Jefferson Lab groups to Summer 2005 Physics Fests Science Education staff member Steve Gagnon Steve Gagnon, Science Education, doing the popular "Deep Freeze (cryogenics)" presentation at the 2003 Open House. Photo credit: Greg Adams. Jefferson Lab invites families, groups to Summer 2005 Physics Fests April 7, 2005 Wednesday, June 8 Wednesday, June 29 Tuesday, July 12 Wednesday, July 27 Wednesday, August 10 This two-hour presentation (10 a.m. - noon) includes an interactive summary

  8. Jefferson Lab invites families, groups to Summer Physics Fests | Jefferson

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Lab groups to Summer Physics Fests Jefferson Lab invites families, groups to Summer Physics Fests April 12, 2004 Wednesday, June 9 Wednesday, June 30 Wednesday, July 28 Wednesday, August 11 Wednesday, August 25 This two-hour presentation (10 a.m. - noon) includes an interactive summary of the research conducted at the Department of Energy's Jefferson Lab followed by the popular "Deep Freeze (cryogenics) and Hot Stuff (plasmas)" presentations, which demonstrate some of the

  9. Overexpression, purification and crystallization of the two C-terminal domains of the bifunctional cellulase ctCel9D-Cel44A from Clostridium thermocellum

    SciTech Connect (OSTI)

    Najmudin, Shabir; Guerreiro, Catarina I. P. D.; Ferreira, Luís M. A.; Romão, Maria J. C.; Fontes, Carlos M. G. A.; Prates, José A. M.

    2005-12-01

    The two C-terminal domains of the cellulase ctCel9D-Cel44A from C. thermocellum cellulosome have been crystallized in tetragonal space group P4{sub 3}2{sub 1}2 and X-ray diffraction data have been collected to 2.1 and 2.8 Å from native and seleno-l-methionine-derivative crystals, respectively. Clostridium thermocellum produces a highly organized multi-enzyme complex of cellulases and hemicellulases for the hydrolysis of plant cell-wall polysaccharides, which is termed the cellulosome. The bifunctional multi-modular cellulase ctCel9D-Cel44A is one of the largest components of the C. thermocellum cellulosome. The enzyme contains two internal catalytic domains belonging to glycoside hydrolase families 9 and 44. The C-terminus of this cellulase, comprising a polycystic kidney-disease module (PKD) and a carbohydrate-binding module (CBM44), has been crystallized. The crystals belong to the tetragonal space group P4{sub 3}2{sub 1}2, containing a single molecule in the asymmetric unit. Native and seleno-l-methionine-derivative crystals diffracted to 2.1 and 2.8 Å, respectively.

  10. Impact of Pretreated Switchgrass and Biomass Carbohydrates on Clostridium thermocellum 27405 Cellulosome Composition- a Quantitative Proteomic Analysis

    SciTech Connect (OSTI)

    Raman, Babu; Pan, Chongle; Hurst, Gregory {Greg} B; Rodriguez, Jr., Miguel; McKeown, Catherine K; Lankford, Patricia K; Samatova, Nagiza F; Mielenz, Jonathan R

    2009-01-01

    The anaerobic thermophilic bacterium Clostridium thermocellum is a cellulolytic organism capable of hydrolyzing cellulose and fermenting the hydrolysis products to ethanol and other metabolic products. C. thermocellum achieves efficient cellulose hydrolysis using multiprotein extracellular enzymatic complexes, termed the cellulosomes. In this study, we used quantitative proteomics (multidimensional LC-MS/MS and 15N-metabolic labeling) to measure relative changes in levels of cellulosomal subunit proteins (per CipA scaffoldin basis) when C. thermocellum was grown on a variety of carbon sources [dilute-acid pretreated switchgrass, cellobiose, amorphous cellulose, crystalline cellulose (Avicel) and combinations of crystalline cellulose with pectin or xylan or both]. Cellulosome samples isolated from cultures grown on these carbon sources were compared to 15N labeled cellulosome samples isolated from crystalline cellulose grown cultures. In total from all samples, proteomic analysis identified 59 dockerin- and 8 cohesin-module containing components, including 15 previously undetected cellulosomal subunits. Many cellulosomal components showed differential protein abundance in the presence of non-cellulose substrates in the growth medium. Cellulosome samples from amorphous cellulose, cellobiose and pretreated switchgrass grown cultures displayed the most distinct differences in composition as compared to cellulosome samples from crystalline cellulose grown cultures. While Glycoside Hydrolase Family 9 enzymes showed increased levels in the presence of crystalline cellulose, and pretreated switchgrass in particular, GH5 enzymes showed increased levels in response to the presence of cellulose in general, amorphous or crystalline. Overall, the results suggest a coordinated substrate-specific regulation of cellulosomal composition in C. thermocellum.

  11. PG&E (Gas)- Multi-Family Residential Energy Efficiency Rebates

    Broader source: Energy.gov [DOE]

    Through the Rebates for Multi-Family Properties Program, PG&E offers prescriptive rebates for owners and managers of multi-family properties of two or more units. Boilers, furnaces, clothes...

  12. BC Hydro Brings Energy Savings to Low-Income Families in Canada...

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    BC Hydro Brings Energy Savings to Low-Income Families in Canada BC Hydro Brings Energy Savings to Low-Income Families in Canada The number of British Columbia, Canada, households ...

  13. ETO family protein Mtgr1 mediates Prdm14 functions in stem cell...

    Office of Scientific and Technical Information (OSTI)

    ETO family protein Mtgr1 mediates Prdm14 functions in stem cell maintenance and primordial germ cell formation Citation Details In-Document Search Title: ETO family protein Mtgr1 ...

  14. Structure and T Cell Inhibition Properties of B7 Family Member...

    Office of Scientific and Technical Information (OSTI)

    Structure and T Cell Inhibition Properties of B7 Family Member, B7-H3 Citation Details In-Document Search Title: Structure and T Cell Inhibition Properties of B7 Family Member, ...

  15. STEP-UP Coaches a University Park Family to Comfort and Savings...

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    STEP-UP Coaches a University Park Family to Comfort and Savings STEP-UP Coaches a University Park Family to Comfort and Savings The average American household spends nearly 2,000 ...

  16. Y-12 and East TN Public Broadcasting System ? A Nuclear Family...

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    - A Nuclear Family Video Miniseries The fourth and final episode of A Nuclear Family: Y-12 National Security Complex documentary film miniseries is complete and East TN PBS is...

  17. Energy Auditor - Single Family 2.0: Blower Door Basics | Department of

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Energy Blower Door Basics Energy Auditor - Single Family 2.0: Blower Door Basics Blower Door Basics - Complete (12.5 MB) Lesson Plan: Blower Door Basics (225.08 KB) PowerPoint: Blower Door Basics (12.84 MB) More Documents & Publications Energy Auditor - Single Family 2.0: Mechanical Ventilation Energy Auditor - Single Family 2.0: Zone Pressure Diagnostics Energy Auditor - Single Family 2.0: Building Science Basics

  18. Energy Auditor - Single Family 2.0: Building Shell Retrofit Strategies |

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Department of Energy Building Shell Retrofit Strategies Energy Auditor - Single Family 2.0: Building Shell Retrofit Strategies Building Shell Retrofit Strategies - Complete (3.79 MB) Lesson Plan: Building Shell Retrofit Strategies (320.22 KB) PowerPoint: Building Shell Retrofit Strategies (3.67 MB) More Documents & Publications Energy Auditor - Single Family 2.0: Zone Pressure Diagnostics Energy Auditor - Single Family 2.0: Mobile Home Assessment Energy Auditor - Single Family 2.0: House

  19. Energy Auditor - Single Family 2.0: Mechanical Ventilation | Department of

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Energy Auditor - Single Family 2.0: Mechanical Ventilation Energy Auditor - Single Family 2.0: Mechanical Ventilation Mechanical Ventilation - Complete (22.2 MB) Lesson Plan: Mechanical Ventilation (222.33 KB) PowerPoint: Mechanical Ventilation (22.61 MB) More Documents & Publications Weatherization Installer/Technician Fundamentals 2.0 - Mechanical Ventilation Rough-In Guidelines Energy Auditor - Single Family 2.0: Blower Door Basics Energy Auditor - Single Family 2.0: Mobile Home

  20. Heterologous Expression of Glycosyl Hydrolases in planta: A New...

    Office of Scientific and Technical Information (OSTI)

    Find in Google Scholar Find in Google Scholar Search WorldCat Search WorldCat to find libraries that may hold this journal Have feedback or suggestions for a way to improve these ...

  1. Coastal Maine Botanical Gardens Bosarge Family Education Center

    High Performance Buildings Database

    Boothbay, ME The Bosarge Family Education Center offers the Gardens a unique opportunity to educate, influence and inspire the public regarding environmental sustainable living practices. The building is a new ~8000 SF Education Center adjacent to the existing Visitors Center in Boothbay ME, owned by Coastal Maine Botanical Gardens (CMBG). This project provides an important addition to CMBG's environmental programming and expands the Gardens' public image. It houses administrative office space as well as flexible and adaptable classroom space that can also be used for various events and gatherings.

  2. The development of a prechamber diesel engine family

    SciTech Connect (OSTI)

    Filtri, G.; Morello, L.; Stroppiana, B.

    1989-01-01

    The development of a new family of prechamber diesel engines, based on a technological commonalty with the gasoline engines is reported. The range of diesel engines, all of them four-cylinder-in line, consist of 3 displacements: 1365cc - 1697cc - 1930cc either naturally aspirated or turbocharged. Mention is also made of their most significant technical innovations about their architecture and combustion chambers, and the main components such as block cylinder, head, crankshaft, connecting rods, pistons, timing gear and injection pump control, intake and exhaust manifolds.

  3. Jefferson Lab invites families, groups, to Summer 2006 Physiscs Fests |

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Jefferson Lab groups, to Summer 2006 Physiscs Fests Jefferson Lab invites families, groups to Summer 2006 Physics Fests March 17, 2006 Thursday, June 1 Wednesday, July 12 Wednesday, July 26 A Physics Fest is a two-hour presentation (10 a.m. - noon) that includes an interactive summary of the research conducted at the Department of Energy's Jefferson Lab followed by the popular "Deep Freeze (cryogenics) and Hot Stuff (plasmas)" presentations, which demonstrate some of the technology

  4. Identifying the ionically bound cell wall and intracellular glycoside...

    Office of Scientific and Technical Information (OSTI)

    Find in Google Scholar Find in Google Scholar Search WorldCat Search WorldCat to find libraries that may hold this journal Have feedback or suggestions for a way to improve these ...

  5. Identifying the Ionically Bound Cell Wall and Intracellular Glycoside...

    Office of Scientific and Technical Information (OSTI)

    Stems: Implications for the Genetic Engineering of Bioenergy Crops; Article 315 Title: ... Stems: Implications for the Genetic Engineering of Bioenergy Crops; Article 315 ...

  6. An evaluation of genetic heterogeneity in 145 breast-ovarian cancer families

    SciTech Connect (OSTI)

    Narod, S.A.; Ford, D.; Devilee, P.; Barkardottir, R.B.; Lynch, H.T.; Smith, S.A.; Ponder, B.A.J.; Weber, B.L.; Garber, J.E.; Birch, J.M.

    1995-01-01

    The breast-ovary cancer-family syndrome is a dominant predisposition to cancer of the breast and ovaries which has been mapped to chromosome region 17q12-q21. The majority, but not all, of breast-ovary cancer families show linkage to this susceptibility locus, designated BRCA1. We report the results of a linkage analysis of 145 families with both breast and ovarian cancer. These families contain either a total of three or more cases of early-onset (before age 60 years) breast cancer or ovarian cancer. All families contained at least one case of ovarian cancer. Overall, an estimated 76% of the 145 families are linked to the BRCA1 locus. None of the 13 families with cases of male breast cancer appear to be linked, but it is estimated that 92% (95% confidence interval 76%-100%) of families with no male breast cancer and with two or more ovarian cancers are linked to BRCA1. These data suggest that the breast-ovarian cancer-family syndrome is genetically heterogeneous. However, the large majority of families with early-onset breast cancer and with two or more cases of ovarian cancer are likely to be due to BRCA1 mutations. 39 refs., 6 figs., 3 tabs.

  7. A novel family of small proteins that affect plant development

    SciTech Connect (OSTI)

    John Charles Walker

    2011-04-29

    The DVL genes represent a new group of plant proteins that influence plant growth and development. Overexpression of DVL1, and other members of the DVL family, causes striking phenotypic changes. The DVL proteins share sequence homology in their C-terminal half. Point mutations in the C-terminal domain show it is necessary and deletion studies demonstrate the C-terminal domain is sufficient to confer the overexpression phenotypes. The phenotypes observed, and the conservation of the protein sequence in the plant kingdom, does suggest the DVL proteins have a role in modulating plant growth and development. Our working hypothesis is the DVL proteins function as regulators of cellular signaling pathways that control growth and development.

  8. Palmitoylation of POTE family proteins for plasma membrane targeting

    SciTech Connect (OSTI)

    Das, Sudipto; Ise, Tomoko; Nagata, Satoshi; Maeda, Hiroshi; Bera, Tapan K.; Pastan, Ira

    2007-11-23

    The POTE gene family is composed of 13 paralogs and likely evolved by duplications and remodeling of the human genome. One common property of POTE proteins is their localization on the inner aspect of the plasma membrane. To determine the structural elements required for membrane localization, we expressed mutants of different POTEs in 293T cells as EGFP fusion proteins. We also tested their palmitoylation by a biotin-switch assay. Our data indicate that the membrane localizations of different POTEs are mediated by similar 3-4 short cysteine rich repeats (CRRs) near the amino-terminuses and that palmitoylation on paired cysteine residues in each CRR motif is responsible for the localization. Multiple palmitoylation in the small CRRs can result in the strong association of whole POTEs with plasma membrane.

  9. A new family of salts for lithium secondary batteries

    SciTech Connect (OSTI)

    Baril, D.; Beranger, S.; Ravet, N.; Michot, C.; Armand, M.

    2000-07-01

    A novel family of salts suitable for lithium battery application was synthesized and characterized. These salts have a large delocalized anion whose charge is spread over a single SO{sub 2} and a phenyl ring. Remarkable properties were obtained for the lithium N-(3-trifluoromethyl phenyl) trifluoromethanesulfonamide salt or LiTFPTS. The electrochemical stability window is around 4.0 V and its conductivity in solid poly(ethylene oxide) or PEO is close to the one of the lithium perchlorate salt. Calorimetric analysis also showed that LiTFPTS behaves as a plasticizer since it hinders, to a certain extent, the PEO crystallization when it is used in a solid polymer matrix. Above all, its synthesis is quite straightforward and leads to potentially inexpensive salts as the starting amines are made commercially on a large scale.

  10. Energy Auditor - Single Family 2.0: Mobile Home Assessment | Department of

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Energy Mobile Home Assessment Energy Auditor - Single Family 2.0: Mobile Home Assessment Mobile Home Assessment - Complete (5.85 MB) Lesson Plan: Mobile Home Assessment (310.8 KB) PowerPoint: Mobile Home Assessment (5.69 MB) More Documents & Publications Weatherization Installer/Technician Fundamentals 2.0 - Mechanical Systems and Improve Duct Systems Energy Auditor - Single Family 2.0: Building Shell Retrofit Strategies Energy Auditor - Single Family 2.0: HVAC Distribution System

  11. Linkage analysis of 19 French breast cancer families, with five chromosome 17q markers

    SciTech Connect (OSTI)

    Mazoyer, S.; Jamot, B.; Sobol, H. ); Lalle, P.; Bignon, Y.J.; Courjal, F. ); Narod, S.A. ); Dutrillaux, B.; Stoppa-Lyonnet, D. )

    1993-04-01

    Nineteen French breast and breast-ovarian cancer families were tested for linkage with five chromosome 17q markers. The five breast-ovarian cancer families as a group give positive evidence for linkage, whereas the 14 breast cancer-only families do not. Heterogeneity of linkage of breast and breast-ovarian cancers is significant in France and supports the existence of more than one susceptibility gene. 15 refs., 2 figs., 3 tabs.

  12. Y-12 employees, families and friends help East Tennessee during annual

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Volunteer Day | Y-12 National Security Complex employees, families ... Y-12 employees, families and friends help East Tennessee during annual Volunteer Day Posted: May 7, 2015 - 3:34pm Chris Clark, Contractor Assurance manager, and Rick Glass, Vice President of Mission Assurance for CNS, volunteered at the Oak Ridge Children's Museum. Y-12 National Security Complex employees, their families and friends lent helping hands to some 32 projects throughout the area - painting, landscaping,

  13. Energy Auditor - Single Family 2.0: Energy Movement | Department of Energy

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Energy Movement Energy Auditor - Single Family 2.0: Energy Movement Energy Movement - Complete (3.44 MB) Lesson Plan: Energy Movement (227.1 KB) PowerPoint: Energy Movement (2.86 MB) Quiz: Energy Movement (229.1 KB) Quiz Key: Energy Movement (233.08 KB) More Documents & Publications Energy Auditor - Single Family 2.0: House as a System Energy Auditor - Single Family 2.0: Building Shell Retrofit Strategies Energy Auditor - Single Family 2.0: Mobile Home Assessment

  14. Weatherized Homes Saving Money for Families Across the U.S.

    Broader source: Energy.gov [DOE]

    750 thousand homes have been weatherized over the past three years through the Department's Weatherization Assistance Program, saving families $400 a year on their heating and cooling bills.

  15. "Feds Feed Families" Events to Be Held at Department of Energy Facilities |

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Department of Energy "Feds Feed Families" Events to Be Held at Department of Energy Facilities "Feds Feed Families" Events to Be Held at Department of Energy Facilities August 4, 2011 - 9:23am Addthis Deputy Secretary Daniel Poneman and his band "Yellow Cake" at last year's Feds Feed Families event. | Energy Department photo Deputy Secretary Daniel Poneman and his band "Yellow Cake" at last year's Feds Feed Families event. | Energy Department photo

  16. Pyrin gene and mutants thereof, which cause familial Mediterranean fever

    DOE Patents [OSTI]

    Kastner, Daniel L.; Aksentijevichh, Ivona; Centola, Michael; Deng, Zuoming; Sood, Ramen; Collins, Francis S.; Blake, Trevor; Liu, P. Paul; Fischel-Ghodsian, Nathan; Gumucio, Deborah L.; Richards, Robert I.; Ricke, Darrell O.; Doggett, Norman A.; Pras, Mordechai

    2003-09-30

    The invention provides the nucleic acid sequence encoding the protein associated with familial Mediterranean fever (FMF). The cDNA sequence is designated as MEFV. The invention is also directed towards fragments of the DNA sequence, as well as the corresponding sequence for the RNA transcript and fragments thereof. Another aspect of the invention provides the amino acid sequence for a protein (pyrin) associated with FMF. The invention is directed towards both the full length amino acid sequence, fusion proteins containing the amino acid sequence and fragments thereof. The invention is also directed towards mutants of the nucleic acid and amino acid sequences associated with FMF. In particular, the invention discloses three missense mutations, clustered in within about 40 to 50 amino acids, in the highly conserved rfp (B30.2) domain at the C-terminal of the protein. These mutants include M6801, M694V, K695R, and V726A. Additionally, the invention includes methods for diagnosing a patient at risk for having FMF and kits therefor.

  17. An interactive grid generator for TOUGH family code

    Energy Science and Technology Software Center (OSTI)

    2004-01-09

    WinGridder has been developed for designing, generating, and visualizing (at various spatial scales) numerical grids used in reservoir simulations and groundwater modeling studies. It can save mesh files for TOUGH family codes and output additional grid information for various purposes in either graphic format or plain text format, many important features, such as inclined faults and offset, layering structure, local refinements, and embedded engineering structures, can be represented in the grid. The main advantages ofmore » this grid-generation software are its user friendly graphical interfaces, flexible grid design capabilities, efficient grid generation, and powerful searching and post-processing capability, especially for large size grid (e.g., a grid of million grid cells). The main improvements of the version 2.0 are (1) to add a capability of handling a repository with multiple sub-regions and specified drifts, (2) to use an interpolation method, instead of picking the nearest point, in calculating the geological data from the given digital geological model, and (3) enhanced searching and other capability.« less

  18. Heterogeneity analysis in 40 X-linked retinitis pigmentosa families

    SciTech Connect (OSTI)

    Teague, P.W.; Aldred, M.A.; Dempster, M.; Harrison, C.; Carothers, A.D.; Hardwick, L.J.; Evans, H.J.; Wright, A.F.; Strain, L.; Brock, D.J.H. )

    1994-07-01

    Analysis of genetic heterogeneity in 40 kindreds with X-linked retinitis pigmentosa (XLRP), with 20 polymorphic markers, showed that significant heterogeneity is present (P=.001) and that 56% of kindreds are of RP3 type and that 26% are of RP2 type. The location of the RP3 locus was found to be 0.4 cM distal to OTC in the Xp21.1 region, and that of the RP2 locus was 6.5 cM proximal to DXS7 in Xp11.2-p11.3. Bayesian probabilities of linkage to RP2, RP3, or to neither locus were calculated. This showed that 20 of 40 kindreds could be assigned to one or the other locus, with a probability >.70 (14 kindreds with RP3 and 6 kindreds with RP2 disease). A further three kindreds were found to be unlinked to either locus, with a probability >.8. The remaining 17 kindreds could not be classified unambiguously. This highlights the difficulty of classifying families in the presence of genetic heterogeneity, where two loci are separated by an estimated 16 cM. 34 refs., 1 fig., 4 tabs.

  19. Functional and structural diversity in GH62 ?-L-arabinofuranosidases from the thermophilic fungus Scytalidium thermophilum

    SciTech Connect (OSTI)

    Kaur, Amrit Pal; Nocek, Boguslaw P.; Xu, Xiaohui; Lowden, Michael J.; Leyva, Juan Francisco; Stogios, Peter J.; Cui, Hong; Leo, Rosa Di; Powlowski, Justin; Tsang, Adrian; Savchenko, Alexei

    2015-05-01

    The genome of the thermophilic fungus Scytalidium thermophilum (strain CBS 625.91) harbours a wide range of genes involved in carbohydrate degradation, including three genes, abf62A, abf62B and abf62C, predicted to encode glycoside hydrolase family 62 (GH62) enzymes. Transcriptome analysis showed that only abf62A and abf62C are actively expressed during growth on diverse substrates including straws from barley, alfalfa, triticale and canola. The abf62A and abf62C genes were expressed in Escherichia coli and the resulting recombinant proteins were characterized. Calcium-free crystal structures of Abf62C in apo and xylotriose bound forms were determined to 1.23 and 1.48 resolution respectively. Site-directed mutagenesis confirmed Asp55, Asp171 and Glu230 as catalytic triad residues, and revealed the critical role of non-catalytic residues Asp194, Trp229 and Tyr338 in positioning the scissile ?-L-arabinofuranoside bond at the catalytic site. Further, the +2R substrate-binding site residues Tyr168 and Asn339, as well as the +2NR residue Tyr226, are involved in accommodating long-chain xylan polymers. Overall, our structural and functional analysis highlights characteristic differences between Abf62A and Abf62C, which represent divergent subgroups in the GH62 family.

  20. Where are the mini Kreutz-family comets?

    SciTech Connect (OSTI)

    Ye, Quan-Zhi; Wiegert, Paul A.; Hui, Man-To; Kracht, Rainer

    2014-12-01

    The Kreutz family of sungrazing comets contains over 2000 known members, many of which are believed to be under ∼100 m sizes (mini comets) and have only been studied at small heliocentric distances (r {sub H}) with space-based SOHO/STEREO spacecraft. To understand the brightening process of mini Kreutz comets, we conducted a survey using CFHT/MegaCam at moderate r {sub H} guided by SOHO/STEREO observations. We identify two comets that should be in our search area but are not detected, indicating that the comets have either followed a steeper brightening rate within the previously reported rapid brightening stage (the brightening burst), or the brightening burst starts earlier than expected. We present a composite analysis of the pre-perihelion light curves of five Kreutz comets that cover to ∼1 AU. We observe significant diversity in the light curves that can be used to grossly classify them into two types: C/Ikeya-Seki and C/SWAN follow the canonical r{sub H}{sup −4} while the others follow r{sub H}{sup −7}. In particular, C/SWAN seems to have undergone an outburst (Δm > 5 mag) or a rapid brightening (n ≳ 11) between r {sub H} = 1.06 AU and 0.52 AU, and shows hints of structural/compositional differences compared to other bright Kreutz comets. We also find evidence that the Kreutz comets as a population lose their mass less efficiently than the dynamically new comet, C/ISON, and are relatively devoid of species that drive C/ISON's activity at large r {sub H}. Concurrent observations of C/STEREO in different wavelengths also suggest that a blueward species such as CN may be the main driver for brightening bursts, instead of sodium as previously thought.

  1. PATtyFams: Protein families for the microbial genomes in the PATRIC database

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Davis, James J.; Gerdes, Svetlana; Olsen, Gary J.; Olson, Robert; Pusch, Gordon D.; Shukla, Maulik; Vonstein, Veronika; Wattam, Alice R.; Yoo, Hyunseung

    2016-02-08

    The ability to build accurate protein families is a fundamental operation in bioinformatics that influences comparative analyses, genome annotation, and metabolic modeling. For several years we have been maintaining protein families for all microbial genomes in the PATRIC database (Pathosystems Resource Integration Center, patricbrc.org) in order to drive many of the comparative analysis tools that are available through the PATRIC website. However, due to the burgeoning number of genomes, traditional approaches for generating protein families are becoming prohibitive. In this report, we describe a new approach for generating protein families, which we call PATtyFams. This method uses the k-mer-based functionmore » assignments available through RAST (Rapid Annotation using Subsystem Technology) to rapidly guide family formation, and then differentiates the function-based groups into families using a Markov Cluster algorithm (MCL). In conclusion, this new approach for generating protein families is rapid, scalable and has properties that are consistent with alignment-based methods.« less

  2. Tactical Response Force 1 graduation: a family reunion of sorts | Y-12

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    National Security Complex Tactical Response Force 1 ... Tactical Response Force 1 graduation: a family reunion of sorts Posted: July 28, 2016 - 2:51pm Tactical Response Force 1 Class 3-16. "Family ties" makes a fitting theme for Tactical Response Force 1 Class 3-16's graduation ceremony. Three kinds of family attended the graduation at New Hope Center: graduates' proud parents, spouses and children; CNS employees watching their children or other relatives graduate; and the

  3. PG&E- Multi-Family Residential Energy Efficiency Rebate Program

    Broader source: Energy.gov [DOE]

    PG&E offers prescriptive rebates for owners and managers of multi-family properties of two or more units. Appliances, boilers, water heating, HVAC, and lighting improvements are among the...

  4. Table 5.8. U.S. Vehicle Fuel Consumption by Family Income, 1994

    U.S. Energy Information Administration (EIA) Indexed Site

    and 1994 Vehicle Characteristics RSE Column Factor: Total 1993 Family Income Below Poverty Line Eli- gible for Fed- eral Assist- ance 1 RSE Row Factor: Less than 5,000 5,000...

  5. Table 5.10. U.S. Average Vehicle Fuel Consumption by Family...

    U.S. Energy Information Administration (EIA) Indexed Site

    1993 Household Characteristics RSE Column Factor: Total 1993 Family Income Below Poverty Line Eli- gible for Fed- eral Assist- ance 1 RSE Row Factor: Less than 5,000 5,000...

  6. Table 5.9. U.S. Average Vehicle-Miles Traveled by Family Income...

    U.S. Energy Information Administration (EIA) Indexed Site

    1993 Household Characteristics RSE Column Factor: Total 1993 Family Income Below Poverty Line Eli- gible for Fed- eral Assist- ance 1 RSE Row Factor: Less than 5,000 5,000...

  7. Working Families Tax Relief Act of 2004 (released in AEO2005)

    Reports and Publications (EIA)

    2005-01-01

    The Working Families Tax Relief Act of 2004 was signed into law on October 13, 2004. Primarily, the Act reduces taxes for individuals and businesses. At least two provisions relate to energy: Depletion of marginal properties and qualified vehicles.

  8. Guidelines for Home Energy Professionals: Standard Work Specifications for Single-Family Home Energy Upgrades

    Broader source: Energy.gov [DOE]

    The Standard Work Specifications (SWS) for Single-Family Home Energy Upgrades define the minimum acceptable outcomes for any weatherization or home performance task to be effective, durable, and safe.

  9. Roy Williams as recalled by his son and family ? Or: Roy Williams...

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    his son and family - Or: Roy Williams: A leader among many (title at it appeared The Oak Ridger) The rewards for writing stories about Y-12 history come in many and varied formsl...

  10. A Nuclear Family: Extras - About the Series | Y-12 National Security...

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    - ... A Nuclear Family: Extras - About the Series The mp4 video format is not supported by this browser. Download video Captions: On Time: 11:49 min. D. Ray Smith, Y-12 historian,...

  11. A Nuclear Family: I've Seen It | Y-12 National Security Complex

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    I've ... A Nuclear Family: I've Seen It The mp4 video format is not supported by this browser. Download video Captions: On Time: 26:42 min. Episode 1 shows how the lives of East...

  12. A Nuclear Family: Lifting The Veil | Y-12 National Security Complex

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Lifting ... A Nuclear Family: Lifting The Veil The mp4 video format is not supported by this browser. Download video Captions: On Time: 26:40 min. Episode 4 describes Y-12's focus...

  13. A Nuclear Family: A Race For Peace | Y-12 National Security Complex

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    A Race ... A Nuclear Family: A Race For Peace The mp4 video format is not supported by this browser. Download video Captions: On Time: 26:40 min. Episode 3 reveals how Y-12 helped...

  14. Zero Energy-Ready Single-Family Homes - Building America Top...

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    View other Top Innovations in the House-as-a-System Business Case category. Zero Energy-Ready Single-Family Homes (766.18 KB) More Documents & Publications Building America Top ...

  15. Seattle City Light- Multi-Family Residential Energy Efficiency Rebate Program

    Broader source: Energy.gov [DOE]

    Seattle City Light provides incentives for multi-family housing properties with 5 or more units to increase their energy efficiency. Rebates are offered for common area lighting and weatherization...

  16. NNSA Production Office donates 50,177 pounds of food for Feds Feed Families

    National Nuclear Security Administration (NNSA)

    campaign | National Nuclear Security Administration | (NNSA) NNSA Production Office donates 50,177 pounds of food for Feds Feed Families campaign September 09, 2016 Amarillo, Texas -- Employees of the National Nuclear Security Administration Production Office (NPO) have donated 50,177pounds of food as part of the annual U.S. Department of Energy's Feds Feed Families (FFF) campaign. The amount of food donated is almost double the amount donated in the 2015 campaign and continues NPO's

  17. The Standard Work Specifications for Single-Family Home Energy Upgrades are

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    now available€ at your fingertips! | Department of Energy The Standard Work Specifications for Single-Family Home Energy Upgrades are now available€ at your fingertips! The Standard Work Specifications for Single-Family Home Energy Upgrades are now available€ at your fingertips! This announcement contains information on the availability of the SWS Online Tool. sws_tool_available.pdf (121.85 KB) More Documents & Publications Guidelines for Home Energy Professionals: Request for

  18. BC Hydro Brings Energy Savings to Low-Income Families in Canada |

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Department of Energy BC Hydro Brings Energy Savings to Low-Income Families in Canada BC Hydro Brings Energy Savings to Low-Income Families in Canada The number of British Columbia, Canada, households eligible for Better Buildings Residential Network member BC Hydro's Energy Conservation Assistance Program (ECAP) just doubled. British Columbia Energy Minister Bill Bennett recently announced an increase in the low-income qualification cutoff for BC Hydro's free home energy-saving kits and

  19. Critical Question #7: What are the Best Practices for Single-Family

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Ventilation in All Climate Regions? | Department of Energy 7: What are the Best Practices for Single-Family Ventilation in All Climate Regions? Critical Question #7: What are the Best Practices for Single-Family Ventilation in All Climate Regions? Why ventilate? What are the ultimate goals of ventilation requirements in codes and standards? What are the characteristics of an effective ventilation system in new vs. existing construction? What are the risks and solutions associated with

  20. Analysis of Installed Measures and Energy Savings for Single-Family Residential Better Buildings Projects

    SciTech Connect (OSTI)

    Heaney, M.; Polly, B.

    2015-04-30

    This report presents an analysis of data for residential single-family projects reported by 37 organizations that were awarded federal financial assistance (cooperative agreements or grants) by the U.S. Department of Energy’s Better Buildings Neighborhood Program.1 The report characterizes the energy-efficiency measures installed for single-family residential projects and analyzes energy savings and savings prediction accuracy for measures installed in a subset of those projects.

  1. A high incidence of BRCA1 mutations in 20 breast-ovarian cancer families

    SciTech Connect (OSTI)

    Serova, O.; Montagna, M.; Sylla, B.

    1996-01-01

    We have analyzed 20 breast-ovarian cancer families, the majority of which show positive evidence of linkage to chromosome 17q12, for germ-line mutations in the BRCA1 gene. BRCA1 mutations cosegregating with breast and ovarian cancer susceptibility were identified in 16 families, including 1 family with a case of male breast cancer. Nine of these mutations have not been reported previously. The majority of mutations were found to generate a premature stop codon leading to the formation of a truncated BRCA1 protein of 2%-88% of the expected normal length. Two mutations altered the RING finger domain. Sequencing of genomic DNA led to the identification of a mutation in the coding region of BRCA1 in 12 families, and cDNA analysis revealed an abnormal or missing BRCA1 transcript in 4 of the 8 remaining families. A total of eight mutations were associated with a reduced quantity of BRCA1 transcript. We were unable to detect BRCA1 mutations in 4 of the 20 families, but only 1 of these was clearly linked to BRCA1. It is expected that the majority of clear examples of the breast-ovarian cancer syndrome will be associated with germ-line mutations in the coding region of BRCA1. 30 refs., 4 figs., 3 tabs.

  2. An estimated 5% of new protein structures solved today represent a new Pfam family

    SciTech Connect (OSTI)

    Mistry, Jaina; Kloppmann, Edda; Rost, Burkhard; Punta, Marco

    2013-11-01

    This study uses the Pfam database to show that the sequence redundancy of protein structures deposited in the PDB is increasing. The possible reasons behind this trend are discussed. High-resolution structural knowledge is key to understanding how proteins function at the molecular level. The number of entries in the Protein Data Bank (PDB), the repository of all publicly available protein structures, continues to increase, with more than 8000 structures released in 2012 alone. The authors of this article have studied how structural coverage of the protein-sequence space has changed over time by monitoring the number of Pfam families that acquired their first representative structure each year from 1976 to 2012. Twenty years ago, for every 100 new PDB entries released, an estimated 20 Pfam families acquired their first structure. By 2012, this decreased to only about five families per 100 structures. The reasons behind the slower pace at which previously uncharacterized families are being structurally covered were investigated. It was found that although more than 50% of current Pfam families are still without a structural representative, this set is enriched in families that are small, functionally uncharacterized or rich in problem features such as intrinsically disordered and transmembrane regions. While these are important constraints, the reasons why it may not yet be time to give up the pursuit of a targeted but more comprehensive structural coverage of the protein-sequence space are discussed.

  3. miR-92a family and their target genes in tumorigenesis and metastasis

    SciTech Connect (OSTI)

    Li, Molin; Guan, Xingfang; Sun, Yuqiang; Mi, Jun; Shu, Xiaohong; Liu, Fang; Li, Chuangang

    2014-04-15

    The miR-92a family, including miR-25, miR-92a-1, miR-92a-2 and miR-363, arises from three different paralog clusters miR-17-92, miR-106a-363, and miR-106b-25 that are highly conservative in the process of evolution, and it was thought as a group of microRNAs (miRNAs) correlated with endothelial cells. Aberrant expression of miR-92a family was detected in multiple cancers, and the disturbance of miR-92a family was related with tumorigenesis and tumor development. In this review, the progress on the relationship between miR-92a family and their target genes and malignant tumors will be summarized. - Highlights: Aberrant expression of miR-92a, miR-25 and miR-363 can be observed in many kinds of malignant tumors. The expression of miR-92a family is regulated by LOH, epigenetic alteration, transcriptional factors such as SP1, MYC, E2F, wild-type p53 etc. Roles of miR-92a family in tumorigenesis and development: promoting cell proliferation, invasion and metastasis, inhibiting cell apoptosis.

  4. The Sorcerer II Global Ocean Sampling Expedition: Expanding theUniverse of Protein Families

    SciTech Connect (OSTI)

    Yooseph, Shibu; Sutton, Granger; Rusch, Douglas B.; Halpern,Aaron L.; Williamson, Shannon J.; Remington, Karin; Eisen, Jonathan A.; Heidelberg, Karla B.; Manning, Gerard; Li, Weizhong; Jaroszewski, Lukasz; Cieplak, Piotr; Miller, Christopher S.; Li, Huiying; Mashiyama, Susan T.; Joachimiak, Marcin P.; van Belle, Christopher; Chandonia, John-Marc; Soergel, David A.; Zhai, Yufeng; Natarajan, Kannan; Lee, Shaun; Raphael,Benjamin J.; Bafna, Vineet; Friedman, Robert; Brenner, Steven E.; Godzik,Adam; Eisenberg, David; Dixon, Jack E.; Taylor, Susan S.; Strausberg,Robert L.; Frazier, Marvin; Venter, J.Craig

    2006-03-23

    Metagenomics projects based on shotgun sequencing of populations of micro-organisms yield insight into protein families. We used sequence similarity clustering to explore proteins with a comprehensive dataset consisting of sequences from available databases together with 6.12 million proteins predicted from an assembly of 7.7 million Global Ocean Sampling (GOS) sequences. The GOS dataset covers nearly all known prokaryotic protein families. A total of 3,995 medium- and large-sized clusters consisting of only GOS sequences are identified, out of which 1,700 have no detectable homology to known families. The GOS-only clusters contain a higher than expected proportion of sequences of viral origin, thus reflecting a poor sampling of viral diversity until now. Protein domain distributions in the GOS dataset and current protein databases show distinct biases. Several protein domains that were previously categorized as kingdom specific are shown to have GOS examples in other kingdoms. About 6,000 sequences (ORFans) from the literature that heretofore lacked similarity to known proteins have matches in the GOS data. The GOS dataset is also used to improve remote homology detection. Overall, besides nearly doubling the number of current proteins, the predicted GOS proteins also add a great deal of diversity to known protein families and shed light on their evolution. These observations are illustrated using several protein families, including phosphatases, proteases, ultraviolet-irradiation DNA damage repair enzymes, glutamine synthetase, and RuBisCO. The diversity added by GOS data has implications for choosing targets for experimental structure characterization as part of structural genomics efforts. Our analysis indicates that new families are being discovered at a rate that is linear or almost linear with the addition of new sequences, implying that we are still far from discovering all protein families in nature.

  5. Linkage studies with 17q and 18q markers in a breast/ovarian cancer family

    SciTech Connect (OSTI)

    Milner, B.J.; Allan, L.A.; Kelly, K.F.; Johnston, A.; Haites, N. ); Cruickshank, D.; Hall, M.; Kitchener, H.; Parkin, D. )

    1993-04-01

    Genes on chromosomes 17q and 18q have been shown to code for putative tumor suppressors. By a combination of allele-loss studies on sporadic ovarian carcinomas and linkage analysis on a breast/ovarian cancer family, the authors have investigated the involvement of such genes in these diseases. Allele loss occurred in sporadic tumors from both chromosome 17p, in 18/26 (69%) cases, and chromosome 17q, in 15/22 (68%) cases. In the three familial tumors studied, allele loss also occurred on chromosome 17 (in 2/3 cases for 17p markers and in 2/2 cases for a 17q allele). Allele loss on chromosome 18q, at the DCC (deleted in colorectal carcinomas) locus, was not as common (6/16 cases [38%]) in sporadic ovarian tumors but had occurred in all three familial tumors. The results of linkage analysis on the breast/ovarian cancer family suggested linkage between the disease locus and 17q markers, with a maximum lod score of 1.507 obtained with Mfd188 (D17S579) polymorphism at 5% recombination. The maximum lod score for DCC was 0.323 at 0.1% recombination. In this family the results are consistent with a predisposing gene for breast/ovarian cancer being located at chromosome 17q21. 24 refs., 1 fig., 3 tabs.

  6. Relativistic static thin dust disks with an inner edge: An infinite family of new exact solutions

    SciTech Connect (OSTI)

    Gonzalez, Guillermo A.; Gutierrez-Pineres, Antonio C.; Vina-Cervantes, Viviana M.

    2009-06-15

    An infinite family of new exact solutions of the vacuum Einstein equations is presented. The solutions are static and axially symmetric and correspond to an infinite family of thin dust disks with a central inner edge. The metric functions of all the solutions can be explicitly computed, and can be expressed in a simple manner in terms of oblate spheroidal coordinates. The energy density of all the disks of the family is positive everywhere and well behaved, so that the corresponding energy-momentum tensor is in full agreement with all the energy conditions. Moreover, although the total mass of the disks is infinite, the solutions are asymptotically flat and the Riemann tensor is regular everywhere, as it is shown by computing the curvature scalars. Now, besides its importance as a new family of exact solutions of the vacuum Einstein equations, the main importance of this family of solutions is that it can be easily superposed with the Schwarzschild solution in order to describe thin disks surrounding a central black hole. Accordingly, a detailed analysis of this superposition will be presented in a subsequent paper.

  7. Comparative genomics of the white-rot fungi, Phanerochaete carnosa and P. chrysosporium, to elucidate the genetic basis of the distinct wood types they colonize

    SciTech Connect (OSTI)

    Suzuki, Hitoshi; MacDonald, Jacqueline; Syed, Khajamohiddin; Salamov, Asaf; Hori, Chiaki; Aerts, Andrea; Henrissat, Bernard; Wiebenga, Ad; vanKuyk, Patricia A.; Barry, Kerrie; Lindquist, Erika; LaButti, Kurt; Lapidus, Alla; Lucas, Susan; Coutinho, Pedro; Gong, Yunchen; Samejima, Masahiro; Mahadevan, Radhakrishnan; Abou-Zaid, Mamdouh; de Vries, Ronald P.; Igarashi, Kiyohiko; Yadav, Jagit S.; Grigoriev, Igor V.; Master, Emma R.

    2012-02-17

    Background Softwood is the predominant form of land plant biomass in the Northern hemisphere, and is among the most recalcitrant biomass resources to bioprocess technologies. The white rot fungus, Phanerochaete carnosa, has been isolated almost exclusively from softwoods, while most other known white-rot species, including Phanerochaete chrysosporium, were mainly isolated from hardwoods. Accordingly, it is anticipated that P. carnosa encodes a distinct set of enzymes and proteins that promote softwood decomposition. To elucidate the genetic basis of softwood bioconversion by a white-rot fungus, the present study reports the P. carnosa genome sequence and its comparative analysis with the previously reported P. chrysosporium genome. Results P. carnosa encodes a complete set of lignocellulose-active enzymes. Comparative genomic analysis revealed that P. carnosa is enriched with genes encoding manganese peroxidase, and that the most divergent glycoside hydrolase families were predicted to encode hemicellulases and glycoprotein degrading enzymes. Most remarkably, P. carnosa possesses one of the largest P450 contingents (266 P450s) among the sequenced and annotated wood-rotting basidiomycetes, nearly double that of P. chrysosporium. Along with metabolic pathway modeling, comparative growth studies on model compounds and chemical analyses of decomposed wood components showed greater tolerance of P. carnosa to various substrates including coniferous heartwood. Conclusions The P. carnosa genome is enriched with genes that encode P450 monooxygenases that can participate in extractives degradation, and manganese peroxidases involved in lignin degradation. The significant expansion of P450s in P. carnosa, along with differences in carbohydrate- and lignin-degrading enzymes, could be correlated to the utilization of heartwood and sapwood preparations from both coniferous and hardwood species.

  8. Multifunctional cellulase catalysis targeted by fusion to different carbohydrate-binding modules

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Walker, Johnnie A.; Takasuka, Taichi E.; Deng, Kai; Bianchetti, Christopher M.; Udell, Hannah S.; Prom, Ben M.; Kim, Hyunkee; Adams, Paul D.; Northen, Trent R.; Fox, Brian G.

    2015-12-21

    Carbohydrate binding modules (CBMs) bind polysaccharides and help target glycoside hydrolases catalytic domains to their appropriate carbohydrate substrates. To better understand how CBMs can improve cellulolytic enzyme reactivity, representatives from each of the 18 families of CBM found in Ruminoclostridium thermocellum were fused to the multifunctional GH5 catalytic domain of CelE (Cthe_0797, CelEcc), which can hydrolyze numerous types of polysaccharides including cellulose, mannan, and xylan. Since CelE is a cellulosomal enzyme, none of these fusions to a CBM previously existed. CelEcc_CBM fusions were assayed for their ability to hydrolyze cellulose, lichenan, xylan, and mannan. Several CelEcc_CBM fusions showed enhanced hydrolyticmore » activity with different substrates relative to the fusion to CBM3a from the cellulosome scaffoldin, which has high affinity for binding to crystalline cellulose. Additional binding studies and quantitative catalysis studies using nanostructure-initiator mass spectrometry (NIMS) were carried out with the CBM3a, CBM6, CBM30, and CBM44 fusion enzymes. In general, and consistent with observations of others, enhanced enzyme reactivity was correlated with moderate binding affinity of the CBM. Numerical analysis of reaction time courses showed that CelEcc_CBM44, a combination of a multifunctional enzyme domain with a CBM having broad binding specificity, gave the fastest rates for hydrolysis of both the hexose and pentose fractions of ionic-liquid pretreated switchgrass. In conclusion, we have shown that fusions of different CBMs to a single multifunctional GH5 catalytic domain can increase its rate of reaction with different pure polysaccharides and with pretreated biomass. This fusion approach, incorporating domains with broad specificity for binding and catalysis, provides a new avenue to improve reactivity of simple combinations of enzymes within the complexity of plant biomass.« less

  9. Multifunctional cellulase catalysis targeted by fusion to different carbohydrate-binding modules

    SciTech Connect (OSTI)

    Walker, Johnnie A.; Takasuka, Taichi E.; Deng, Kai; Bianchetti, Christopher M.; Udell, Hannah S.; Prom, Ben M.; Kim, Hyunkee; Adams, Paul D.; Northen, Trent R.; Fox, Brian G.

    2015-12-21

    Carbohydrate binding modules (CBMs) bind polysaccharides and help target glycoside hydrolases catalytic domains to their appropriate carbohydrate substrates. To better understand how CBMs can improve cellulolytic enzyme reactivity, representatives from each of the 18 families of CBM found in Ruminoclostridium thermocellum were fused to the multifunctional GH5 catalytic domain of CelE (Cthe_0797, CelEcc), which can hydrolyze numerous types of polysaccharides including cellulose, mannan, and xylan. Since CelE is a cellulosomal enzyme, none of these fusions to a CBM previously existed. CelEcc_CBM fusions were assayed for their ability to hydrolyze cellulose, lichenan, xylan, and mannan. Several CelEcc_CBM fusions showed enhanced hydrolytic activity with different substrates relative to the fusion to CBM3a from the cellulosome scaffoldin, which has high affinity for binding to crystalline cellulose. Additional binding studies and quantitative catalysis studies using nanostructure-initiator mass spectrometry (NIMS) were carried out with the CBM3a, CBM6, CBM30, and CBM44 fusion enzymes. In general, and consistent with observations of others, enhanced enzyme reactivity was correlated with moderate binding affinity of the CBM. Numerical analysis of reaction time courses showed that CelEcc_CBM44, a combination of a multifunctional enzyme domain with a CBM having broad binding specificity, gave the fastest rates for hydrolysis of both the hexose and pentose fractions of ionic-liquid pretreated switchgrass. In conclusion, we have shown that fusions of different CBMs to a single multifunctional GH5 catalytic domain can increase its rate of reaction with different pure polysaccharides and with pretreated biomass. This fusion approach, incorporating domains with broad specificity for binding and catalysis, provides a new avenue to improve reactivity of simple combinations of enzymes within the complexity of plant biomass.

  10. Complete genome of the cellyloytic thermophile Acidothermus cellulolyticus 11B provides insights into its ecophysiological and evloutionary adaptations

    SciTech Connect (OSTI)

    Barabote, Ravi D.; Xie, Gary; Leu, David H.; Normand, Philippe; Necsulea, Anamaria; Daubin, Vincent; Medigue, Claudine; Adney, William S.; Xu,Xin Clare; Lapidus, Alla; Detter, Chris; Pujic, Petar; Bruce, David; Lavire, Celine; Challacombe, Jean F.; Brettin, Thomas S.; Berry, Alison M.

    2009-01-01

    We present here the complete 2.4 Mb genome of the cellulolytic actinobacterial thermophile, Acidothermus cellulolyticus 11B. New secreted glycoside hydrolases and carbohydrate esterases were identified in the genome, revealing a diverse biomass-degrading enzyme repertoire far greater than previously characterized, and significantly elevating the industrial value of this organism. A sizable fraction of these hydrolytic enzymes break down plant cell walls and the remaining either degrade components in fungal cell walls or metabolize storage carbohydrates such as glycogen and trehalose, implicating the relative importance of these different carbon sources. A novel feature of the A. cellulolyticus secreted cellulolytic and xylanolytic enzymes is that they are fused to multiple tandemly arranged carbohydrate binding modules (CBM), from families 2 and 3. Interestingly, CBM3 was found to be always N-terminal to CBM2, suggesting a functional constraint driving this organization. While the catalytic domains of these modular enzymes are either diverse or unrelated, the CBMs were found to be highly conserved in sequence and may suggest selective substrate-binding interactions. For the most part, thermophilic patterns in the genome and proteome of A. cellulolyticus were weak, which may be reflective of the recent evolutionary history of A. cellulolyticus since its divergence from its closest phylogenetic neighbor Frankia, a mesophilic plant endosymbiont and soil dweller. However, ribosomal proteins and non-coding RNAs (rRNA and tRNAs) in A. cellulolyticus showed thermophilic traits suggesting the importance of adaptation of cellular translational machinery to environmental temperature. Elevated occurrence of IVYWREL amino acids in A. cellulolyticus orthologs compared to mesophiles, and inverse preferences for G and A at the first and third codon positions also point to its ongoing thermoadaptation. Additional interesting features in the genome of this cellulolytic, hot

  11. Better Buildings Neighborhood Program Multi-family/Low Income Peer Exchange Call: Strategies to Address Split Incentives in Multi-family Buildings Call Slides and Discussion Summary, April 26, 2012

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    26, 2012 Better Buildings Neighborhood Program Multi- family/Low Income Peer Exchange Call: Strategies to Address Split Incentives in Multi-family Buildings Call Slides and Discussion Summary Agenda * Call Logistics and Attendance  How are you working with owners and tenants on multi-family upgrades? * Program Experience and Lessons:  Michael Croston, Repower Bremerton * Discussion:  What are some of the key challenges for working with rental properties?  What are effective

  12. Homogenization of a thin plate reinforced with periodic families of rigid rods

    SciTech Connect (OSTI)

    Nazarov, Sergei A; Sweers, Guido H; Slutskij, Andrey S

    2011-08-31

    The asymptotics of the solution to the elastic bending problem for a thin plate reinforced with several periodic families of closely spaced but disjoint rods are constructed and justified, the result of homogenization being substantially different from the case when the rods are welded together into a single periodic mesh. The material in the rods is assumed to be appreciably more rigid than that in the plate. An averaged fourth-order differential operator is obtained from summing the nonelliptic operators generated by each of the families of the rods. This operator is shown to be elliptic if and only if the rods from at least two families are nonparallel. As a simplified example, the paper examines a similar stationary heat conduction problem. Bibliography: 24 titles.

  13. A new method of calibration and normalization for neutron detector families

    SciTech Connect (OSTI)

    Menlove, H.O.; Stewart, J.E.

    1988-04-01

    A calibration and cross-reference is presented for passive and active neutron assay instruments. The method reduces and number of physical standards required to calibrate families of neutron detectors and also ties together much of the calibration information currently available. The basic approach is to carefully calibrate one member of the family (reference detector) over the complete mass range of interest. Other members of the family can be cross-referenced to the calibrated detector using a single sample or radioactive source. Calibration and cross-reference information is presented for the Inventory Sample Counter, High-Level Neutron Coincidence Counter. Active Well Coincidence Counter, and the Neutron Collar. 1 ref., 15 figs., 20 tabs.

  14. Discrete family of dissipative soliton pairs in mode-locked fiber lasers

    SciTech Connect (OSTI)

    Zavyalov, Aleksandr; Iliew, Rumen; Egorov, Oleg; Lederer, Falk

    2009-05-15

    We numerically investigate the formation of soliton pairs (bound states) in mode-locked fiber ring lasers. In the distributed model (complex cubic-quintic Ginzburg-Landau equation) we observe a discrete family of soliton pairs with equidistantly increasing peak separation. This family was identified by two alternative numerical schemes and the bound state instability was disclosed by a linear stability analysis. Moreover, similar families of unstable bound state solutions have been found in a more realistic lumped laser model with an idealized saturable absorber (instantaneous response). We show that a stabilization of these bound states can be achieved when the finite relaxation time of the saturable absorber is taken into account. The domain of stability can be controlled by varying this relaxation time.

  15. Familial site-specific Ovarian cancer is linked to BRCA1 on 17q12-21

    SciTech Connect (OSTI)

    Steichen-Gersdorf, E.; Gallion, H.H.; Ponder, M.A.; Pye, C.; Mazoyer, S.; Smith, S.A.; Ponder, B.A.J.; Ford, D.; Easton, D.F.; Girodet, C.

    1994-11-01

    In a study of nine families with {open_quotes}site-specific{close_quotes} ovarian cancer (criterion: three or more cases of epithelial ovarian cancer and no cases of breast cancer diagnosed at age <50 years) we have obtained evidence of linkage to the breast-ovarian cancer susceptibility gene, BRCA1 on 17q12-21. If the risk of cancer in these families is assumed to be restricted to the ovary, the best estimate of the proportion of families linked to BRCA1 is .78 (95% confidence interval .32-1.0). If predisposition to both breast and ovarian cancer is assumed, the proportion linked is 1.0 (95% confidence interval .46-1.0). The linkage of familial site-specific ovarian cancer to BRCA1 indicates the possibility of predictive testing in such families; however, this is only appropriate in families where the evidence for linkage to BRCA1 is conclusive. 17 refs., 3 figs., 1 tab.

  16. Energy Department Provides $140.3 Million to Low-Income Families for Home

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Weatherization | Department of Energy 140.3 Million to Low-Income Families for Home Weatherization Energy Department Provides $140.3 Million to Low-Income Families for Home Weatherization April 3, 2006 - 9:55am Addthis Funding is first installment of $243 million in total weatherization grants for FY 2006 WASHINGTON, D.C. - U.S. Department of Energy (DOE) Secretary Samuel W. Bodman today announced $140.3 million in weatherization program grants to 31 states and the Navajo Nation to make

  17. NNSA Production Office tops Feds Feed Families campaign goal | Y-12

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    National Security Complex tops ... NNSA Production Office tops Feds Feed Families campaign goal Posted: September 16, 2013 - 9:45am Oak Ridge, Tenn. - Employees of the National Nuclear Security Administration Production Office have donated 17,348 pounds of food as part of the annual U.S. Department of Energy's "Feds Feed Families" campaign. The campaign, which ended on September 6, surpassed the goal of 17,000 pounds. "I am extremely proud and moved by the generosity of NPO

  18. Department of Energy Provides Nearly $112 Million to Low-Income Families

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    for Home Weatherization | Department of Energy 112 Million to Low-Income Families for Home Weatherization Department of Energy Provides Nearly $112 Million to Low-Income Families for Home Weatherization March 29, 2007 - 12:17pm Addthis Funding is First Installment of $204.5 Million in Total Weatherization Grants for FY 2007 WASHINGTON, DC - U.S. Department of Energy (DOE) today announced $111.6 million in weatherization grants to 30 states and the Navajo Nation to make energy efficiency

  19. Department of Energy Provides Nearly $88 Million to Low-Income Families for

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Home Weatherization | Department of Energy 88 Million to Low-Income Families for Home Weatherization Department of Energy Provides Nearly $88 Million to Low-Income Families for Home Weatherization June 29, 2007 - 2:36pm Addthis Funding is Second Installment of $200 Million in Total Weatherization Grants for FY 2007 WASHINGTON, DC - U.S. Department of Energy (DOE) today announced $88 million in weatherization grants to 20 states to make energy efficiency improvements in homes of low-income

  20. Pantexans donate more than $6,000 of cookware to families in need |

    National Nuclear Security Administration (NNSA)

    National Nuclear Security Administration | (NNSA) donate more than $6,000 of cookware to families in need Thursday, April 3, 2014 - 10:00am Cooking dinner will be a lot easier for about 20 needy families in Amarillo thanks to the generosity of Pantexans, who used a grocery store promotion to secure more than $6,000 of cookware for a local charity. More than 70 pots and pans were donated to local charities by Pantexans Scott and Lauri Minton, who had been collecting stickers donated by their

  1. An Invitation for Family Fun at PPPL's Open House on June 1 | Princeton

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Plasma Physics Lab An Invitation for Family Fun at PPPL's Open House on June 1 By Jeanne Jackson DeVoe May 13, 2013 Tweet Widget Google Plus One Share on Facebook An Invitation for Family Fun at PPPL's Open House on June 1 The U.S. Department of Energy's Princeton Plasma Physics Lab Open House will be a fun science festival for every age, with NASA moon rocks, a Hall of Machines, an art show of paintings of PPPL, science activities for kids and tours of fusion machines. The event on June 1

  2. DOE Provides $96.4 Million to Low-Income Families for Home Weatherization |

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Department of Energy $96.4 Million to Low-Income Families for Home Weatherization DOE Provides $96.4 Million to Low-Income Families for Home Weatherization July 6, 2006 - 2:50pm Addthis Funding is Second Installment of $243 Million in Total Weatherization Grants for FY 2006 WASHINGTON, D.C. - U.S. Department of Energy (DOE) Secretary Samuel W. Bodman today announced $96.4 million in weatherization program grants to 19 states to make energy efficiency improvements in homes of low-income

  3. 10,000th Waste Shipment Milestone is All in the Family | Department of

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Energy Waste Shipment Milestone is All in the Family 10,000th Waste Shipment Milestone is All in the Family August 29, 2011 - 12:00pm Addthis A Waste Isolation Pilot Plant facility shift manager inspects the shipping manifest of the 10,000th safe shipment to roll through the gates at the Department of Energy’s Waste Isolation Pilot Plant on Sept. 24, 2011. A Waste Isolation Pilot Plant facility shift manager inspects the shipping manifest of the 10,000th safe shipment to roll through

  4. Genetic heterogeneity and localization of a familial breast-ovarian cancer gene on chromosome 17q12-q21

    SciTech Connect (OSTI)

    Smith, S.A.; Ponder, M.; Pye, C.; Ponder, B.A.J. ); Easton, D.F.; Ford, D.; Peto, J.; Anderson, K.; Averill, D.; Stratton, M. )

    1993-04-01

    In a study of 31 breast cancer families and 12 breast-ovarian cancer families, we have obtained clear evidence of linkage to markers on chromosome 17q in the families with ovarian cancer (maximum lod score 3.34 at [theta] = .04) but only weak evidence in those without ovarian cancer. Recombinant events indicate that the gene lies between D17S588 and D17S250. 9 refs., 2 figs., 4 tabs.

  5. Implementation of the Research Performance Progress Report (RPPR) and the SF-428 Tangible Personal Property Report Forms Family

    Broader source: Energy.gov [DOE]

    The new Research Performance Progress Report (RPPR) and SF-428 Tangible Personal Property Report Forms Family are now available for use.

  6. The Fokker-Planck limit of a family of transport differencing methods

    SciTech Connect (OSTI)

    Anistratov, D.Y.

    1998-12-31

    Recently, Pomraning performed an asymptotic analysis of the Fokker-Planck (FP) limit for the analytic transport equation with a forward-peaked scattering kernel. Then, Adams and Pautz extended this analysis to the discrete ordinates transport equation and studied some difference schemes. In this paper a broad family of transport differencing methods is analyzed.

  7. Weatherization and Indoor Air Quality: Measured Impacts in Single Family Homes Under the Weatherization Assistance Program

    SciTech Connect (OSTI)

    Pigg, Scott; Cautley, Dan; Francisco, Paul; Hawkins, Beth A; Brennan, Terry M

    2014-09-01

    This report summarizes findings from a national field study of indoor air quality parameters in homes treated under the Weatherization Assistance Program (WAP). The study involved testing and monitoring in 514 single-family homes (including mobile homes) located in 35 states and served by 88 local weatherization agencies.

  8. Do Babies Matter? The Effect of Family Formation on Men and Women in Science

    SciTech Connect (OSTI)

    Mary Ann Mason

    2007-10-30

    Mary Ann Mason, Professor of Social Welfare and Law at the University of California, Berkeley, presents "Do Babies Matter? The Effect of Family Formation on Men and Women in Science." In her talk, she discusses the difficulties of women who have a career in science or in other male-dominated professions.

  9. Do Babies Matter? The Effect of Family Formation on Men and Women in Science

    ScienceCinema (OSTI)

    Mary Ann Mason

    2010-09-01

    Mary Ann Mason, Professor of Social Welfare and Law at the University of California, Berkeley, presents "Do Babies Matter? The Effect of Family Formation on Men and Women in Science." In her talk, she discusses the difficulties of women who have a career in science or in other male-dominated professions.

  10. West Valley Demonstration Project Food Drive Delivers Food for 700 Families

    Office of Energy Efficiency and Renewable Energy (EERE)

    WEST VALLEY, N.Y. – EM employees at West Valley Demonstration Project (WVDP) helped collect and deliver 114,843 pounds of food, including 360 turkeys, to nine food pantries in the West Valley area, just in time to benefit about 700 families in need during the holidays.

  11. Friedreich ataxia in Acadian families from eastern Canada: Clinical diversity with conserved haplotypes

    SciTech Connect (OSTI)

    Richter, A.; Poirier, J.; Mercier, J.

    1996-09-06

    The gene for Friedreich ataxia (FRDA), an autosomal-recessive neurodegenerative disease, remains elusive. The current candidate region of about 150 kb lies between loci FR2 and F8101 near the D9S15/D9S5 linkage group at 9q13-21.1. Linkage homogeneity between classical FRDA and a milder, slowly progressive Acadian variant (FRDA-Acad) has been demonstrated. An extended D9S15-D9S5 haplotype (C6) predominates in FRDA-Acad chromosomes from Louisiana. We studied 10 Acadian families from New Brunswick, Canada. In eight families, affected individuals conformed to the clinical description of FRDA-Acad; in one, 2 sibs presented with spastic ataxia (SPA-Acad). In the last family, 2 sibs had FRDA-Acad, and one had SPA-Acad. We found that SPA-Acad is linked to the FRDA gene region. The C6 haplotype and a second major haplotype (137) were identified. The same ataxia-linked haplotypes segregated with both FRDA-Acad and SPA-Acad in two unrelated families. The parental origins of these haplotypes were different. Our observation of different phenotypes associated with the same combination of haplotypes may point to the influence of the parent of origin on gene expression, indicate the effect of modifier genes, or reflect the presence of different mutations on the same haplotype. Our findings underline the need to investigate families with autosomal-recessive ataxias for linkage to the FRDA region, despite lack of key diagnostic manifestations such as cardiomyopathy or absent deep-tendon reflexes. 28 refs., 4 figs., 3 tabs.

  12. Genome-wide analysis of the omega-3 fatty acid desaturase gene family in Gossypium

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Yurchenko, Olga P.; Park, Sunjung; Ilut, Daniel C.; Inmon, Jay J.; Millhollon, Jon C.; Liechty, Zach; Page, Justin T.; Jenks, Matthew A.; Chapman, Kent D.; Udall, Joshua A.; et al

    2014-11-18

    The majority of commercial cotton varieties planted worldwide are derived from Gossypium hirsutum, which is a naturally occurring allotetraploid produced by interspecific hybridization of A- and D-genome diploid progenitor species. While most cotton species are adapted to warm, semi-arid tropical and subtropical regions, and thus perform well in these geographical areas, cotton seedlings are sensitive to cold temperature, which can significantly reduce crop yields. One of the common biochemical responses of plants to cold temperatures is an increase in omega-3 fatty acids, which protects cellular function by maintaining membrane integrity. The purpose of our study was to identify and characterizemore » the omega-3 fatty acid desaturase (FAD) gene family in G. hirsutum, with an emphasis on identifying omega-3 FADs involved in cold temperature adaptation. Results: Eleven omega-3 FAD genes were identified in G. hirsutum, and characterization of the gene family in extant A and D diploid species (G. herbaceum and G. raimondii, respectively) allowed for unambiguous genome assignment of all homoeologs in tetraploid G. hirsutum. The omega-3 FAD family of cotton includes five distinct genes, two of which encode endoplasmic reticulum-type enzymes (FAD3-1 and FAD3-2) and three that encode chloroplast-type enzymes (FAD7/8-1, FAD7/8-2, and FAD7/8-3). The FAD3-2 gene was duplicated in the A genome progenitor species after the evolutionary split from the D progenitor, but before the interspecific hybridization event that gave rise to modern tetraploid cotton. RNA-seq analysis revealed conserved, gene-specific expression patterns in various organs and cell types and semi-quantitative RT-PCR further revealed that FAD7/8-1 was specifically induced during cold temperature treatment of G. hirsutum seedlings. Conclusions: The omega-3 FAD gene family in cotton was characterized at the genome-wide level in three species, showing relatively ancient establishment of the gene family prior

  13. The earth-coupled heat pump: Utilizing innovative technology in single family rehabilitation strategies

    SciTech Connect (OSTI)

    Not Available

    1989-11-01

    The study examines the feasibility of incorporating the use of earth-coupled heat pump technology in single-family housing rehabilitation projects, based on energy conservation attributes and financial considerations. Following evaluation of a theoretical model which indicated that installations of the heat pumps were feasible, the heat pumps were tested under actual conditions in five single family housing units which were part of the Urban Homesteading Program, and were matched with comparable units which did not receive special treatment. Energy consumption information was collected for all units for twelve months. Variables were identified, and the data was analyzed for individual housing units and compared with the results predicted by the theoretical model to determine the practicality of incorporating such technology in large scale rehabilitation projects. 14 refs., 14 figs., 3 tabs.

  14. Standard Work Specifications for Single-Family Home Energy Upgrades Summary (Fact Sheet)

    SciTech Connect (OSTI)

    Not Available

    2012-11-01

    The U.S. Department of Energy (DOE), the National Renewable Energy Laboratory (NREL) and numerous industry stakeholders developed the Standard Work Specifications for Single-Family Home Energy Upgrades to define the minimum requirements for high-quality residential energy upgrades. Today, the Standard Work Specifications provide a unique source for defining high-quality home energy upgrades, establishing clear expectations for homeowners, contractors, trainers, workers, program administrators, and organizations that provide financing for energy upgrades.

  15. Identifying Inefficient Single-Family Homes With Utility Bill Analysis: Preprint

    SciTech Connect (OSTI)

    Casey, S.; Krarti, M.; Bianchi, M.; Roberts, D.

    2010-08-01

    Differentiating between energy-efficient and inefficient single-family homes on a community scale helps identify and prioritize candidates for energy-efficiency upgrades. Prescreening diagnostic procedures can further retrofit efforts by providing efficiency information before a site-visit is conducted. We applied the prescreening diagnostic to a simulated community of homes in Boulder, Colorado and analyzed energy consumption data to identify energy-inefficient homes.

  16. Inverse scattering method and soliton double solution family for the general symplectic gravity model

    SciTech Connect (OSTI)

    Gao Yajun

    2008-08-15

    A previously established Hauser-Ernst-type extended double-complex linear system is slightly modified and used to develop an inverse scattering method for the stationary axisymmetric general symplectic gravity model. The reduction procedures in this inverse scattering method are found to be fairly simple, which makes the inverse scattering method applied fine and effective. As an application, a concrete family of soliton double solutions for the considered theory is obtained.

  17. Two-dimensional Laguerre-Gaussian soliton family in strongly nonlocal nonlinear media

    SciTech Connect (OSTI)

    Zhong Weiping; Yi Lin

    2007-06-15

    We have studied Laguerre-Gaussian spatial solitary waves in strongly nonlocal nonlinear media analytically and numerically. An exact analytical solution of two-dimensional self-similar waves is obtained. Furthermore, a family of different spatial solitary waves has been found. It is interesting that the spatial soliton profile and its width remain unchanged with increasing propagation distance. The theoretical predictions may give new insights into low-energetic spatial soliton transmission with high fidelity.

  18. Most Complete Functional Map of an Entire Enzyme Family| U.S. DOE Office of

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Science (SC) Most Complete Functional Map of an Entire Enzyme Family Biological and Environmental Research (BER) BER Home About Research Facilities Science Highlights Searchable Archive of BER Highlights External link Benefits of BER Funding Opportunities Biological & Environmental Research Advisory Committee (BERAC) Community Resources Contact Information Biological and Environmental Research U.S. Department of Energy SC-23/Germantown Building 1000 Independence Ave., SW Washington, DC

  19. National impacts of the Weatherization Assistance Program in single-family and small multifamily dwellings

    SciTech Connect (OSTI)

    Brown, M.A.; Berry, L.G.; Balzer, R.A.; Faby, E.

    1993-05-01

    Since 1976, the US Department of Energy (DOE) has operated one of the largest energy conservation programs in the nation -- the low-income Weatherization Assistance Program. The program strives to increase the energy efficiency of dwellings occupied by low-income persons in order to reduce their energy consumption, lower their fuel bills, increase the comfort of their homes, and safeguard their health. It targets vulnerable groups including the elderly, people with disabilities, and families with children. The most recent national evaluation of the impacts of the Program was completed in 1984 based on energy consumption data for households weatherized in 1981. DOE Program regulations and operations have changed substantially since then: new funding sources, management principles, diagnostic procedures, and weatherization technologies have been incorporated. Many of these new features have been studied in isolation or at a local level; however, no recent evaluation has assessed their combined, nationwide impacts to date or their potential for the future. In 1990, DOE initiated such an evaluation. This evaluation is comprised of three ``impact`` studies (the Single-Family Study, High-Density Multifamily Study, and Fuel-Oil Study) and two ``policy`` studies. Altogether, these five studies will provide a comprehensive national assessment of the Weatherization Assistance Program as it existed in the 1989 Program Year (PY 1989). This report presents the results of the first phase of the Single-Family Study. It evaluates the energy savings and cost effectiveness of the Program as it has been applied to the largest portion of its client base -- low-income households that occupy single-family dwellings, mobile homes, and small (2- to 4-unit) multifamily dwellings. It is based upon a representative national sample that covers the full range of conditions under which the program was implemented in PY 1989.

  20. Standard Work Specifications for Single-Family Home Energy Upgrades Summary Fact Sheet

    Broader source: Energy.gov [DOE]

    The U.S. Department of Energy (DOE), the National Renewable Energy Laboratory (NREL) and numerous industry stakeholders developed the Standard Work Specifications for Single-Family Home Energy Upgrades to define the minimum requirements for high-quality residential energy upgrades. Today, the Standard Work Specifications provide a unique source for defining high-quality home energy upgrades, establishing clear expectations for homeowners, contractors, trainers, workers, program administrators, and organizations that provide financing for energy upgrades.

  1. Stockbridge-Munsee Health & Wellness Center and Family Center Renewable Energy Feasibility Study

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Stockbridge-Munsee Health & Wellness Center & Family Center Renewable Energy Feasibility Study. Stockbridge-Munsee Tribal Overview * A Band of Mohican Indians * From the Algonquin language group of Tribes. Papscanee Village Site Painting follows archeological reports & accurately depicts pre European village. Tribal History * The tribe lived along the Muh-he-con-ne-ok River, "where the waters are never still". (now the Hudson River) * The Stockbridge name came from the town

  2. Stockbridge-Munsee Health & Wellness Center and Family Center Renewable Energy Feasibility Study

    Office of Environmental Management (EM)

    Stockbridge-Munsee Health & Wellness Center & Family Center Renewable Energy Feasibility Study. Stockbridge-Munsee Tribal Overview * A Band of Mohican Indians * From the Algonquin language group of Tribes. Papscanee Village Site Painting follows archeological reports & accurately depicts pre European village. Tribal History * The tribe lived along the Muh-he-con-ne-ok River, "where the waters are never still". (now the Hudson River) * The Stockbridge name came from the town

  3. DOE Fact Sheet: Cost-Effectiveness of Deep Green Alterations of Multi-family Buildings in

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Fact Sheet: Cost-Effectiveness of Deep Green Alterations of Multi-family Buildings in Seattle Overview The City of Seattle was a multiple awardee of the Climate Action Champions (CAC) Notice of Technical Assistance (NOTA). The U.S. Department of Energy (DOE)'s Office of Energy Efficiency and Renewable Energy offered technical assistance from its Commercial Building Partnerships (CBP) Program to provide CACs with additional opportunities for technical assistance to support and advance their

  4. Managing the Drivers of Air Flow and Water Vapor Transport in Existing Single-Family Homes

    SciTech Connect (OSTI)

    Cummings, James; Withers, Charles; Martin, Eric; Moyer, Neil

    2012-10-01

    This report is a revision of an earlier report titled: Measure Guideline: Managing the Drivers of Air Flow and Water Vapor Transport in Existing Single-Family Homes. Revisions include: Information in the text box on page 1 was revised to reflect the most accurate information regarding classifications as referenced in the 2012 International Residential Code. “Measure Guideline” was dropped from the title of the report. An addition was made to the reference list.

  5. A new family of β-helix proteins with similarities to the polysaccharide lyases

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Close, Devin W.; D'Angelo, Sara; Bradbury, Andrew R. M.

    2014-09-27

    Microorganisms that degrade biomass produce diverse assortments of carbohydrate-active enzymes and binding modules. Despite tremendous advances in the genomic sequencing of these organisms, many genes do not have an ascribed function owing to low sequence identity to genes that have been annotated. Consequently, biochemical and structural characterization of genes with unknown function is required to complement the rapidly growing pool of genomic sequencing data. A protein with previously unknown function (Cthe_2159) was recently isolated in a genome-wide screen using phage display to identify cellulose-binding protein domains from the biomass-degrading bacterium Clostridium thermocellum. Here, the crystal structure of Cthe_2159 is presentedmore » and it is shown that it is a unique right-handed parallel β-helix protein. Despite very low sequence identity to known β-helix or carbohydrate-active proteins, Cthe_2159 displays structural features that are very similar to those of polysaccharide lyase (PL) families 1, 3, 6 and 9. Cthe_2159 is conserved across bacteria and some archaea and is a member of the domain of unknown function family DUF4353. This suggests that Cthe_2159 is the first representative of a previously unknown family of cellulose and/or acid-sugar binding β-helix proteins that share structural similarities with PLs. More importantly, these results demonstrate how functional annotation by biochemical and structural analysis remains a critical tool in the characterization of new gene products.« less

  6. A new family of β-helix proteins with similarities to the polysaccharide lyases

    SciTech Connect (OSTI)

    Close, Devin W.; D'Angelo, Sara; Bradbury, Andrew R. M.

    2014-09-27

    Microorganisms that degrade biomass produce diverse assortments of carbohydrate-active enzymes and binding modules. Despite tremendous advances in the genomic sequencing of these organisms, many genes do not have an ascribed function owing to low sequence identity to genes that have been annotated. Consequently, biochemical and structural characterization of genes with unknown function is required to complement the rapidly growing pool of genomic sequencing data. A protein with previously unknown function (Cthe_2159) was recently isolated in a genome-wide screen using phage display to identify cellulose-binding protein domains from the biomass-degrading bacterium Clostridium thermocellum. Here, the crystal structure of Cthe_2159 is presented and it is shown that it is a unique right-handed parallel β-helix protein. Despite very low sequence identity to known β-helix or carbohydrate-active proteins, Cthe_2159 displays structural features that are very similar to those of polysaccharide lyase (PL) families 1, 3, 6 and 9. Cthe_2159 is conserved across bacteria and some archaea and is a member of the domain of unknown function family DUF4353. This suggests that Cthe_2159 is the first representative of a previously unknown family of cellulose and/or acid-sugar binding β-helix proteins that share structural similarities with PLs. More importantly, these results demonstrate how functional annotation by biochemical and structural analysis remains a critical tool in the characterization of new gene products.

  7. Retrofit of a MultiFamily Mass Masonry Building in New England

    SciTech Connect (OSTI)

    Ueno, K.; Kerrigan, P.; Wytrykowska, H.; Van Straaten, R.

    2013-08-01

    Merrimack Valley Habitat for Humanity (MVHfH) has partnered with Building Science Corporation to provide high performance affordable housing for 10 families in the retrofit of an existing brick building (a former convent) into condominiums. The research performed for this project provides information regarding advanced retrofit packages for multi-family masonry buildings in Cold climates. In particular, this project demonstrates safe, durable, and cost-effective solutions that will potentially benefit millions of multi-family brick buildings throughout the East Coast and Midwest (Cold climates). The retrofit packages provide insight on the opportunities for and constraints on retrofitting multifamily buildings with ambitious energy performance goals but a limited budget. The condominium conversion project will contribute to several areas of research on enclosures, space conditioning, and water heating. Enclosure items include insulation of mass masonry building on the interior, airtightness of these types of retrofits, multi-unit building compartmentalization, window selection, and roof insulation strategies. Mechanical system items include combined hydronic and space heating systems with hydronic distribution in small (low load) units, and ventilation system retrofits for multifamily buildings.

  8. HUD rehabilitation energy guidelines for one-to-four family dwellings

    SciTech Connect (OSTI)

    1996-09-01

    The Guidebook has been prepared to help people who are involved in the rehabilitation of a single-family house (or low-rise building with up to four housing units). It helps to understand the basics of residential energy conservation, and to think positively about the potential economic benefits. The Guidebook, and the HUD Rehabilitation Energy Guidelines are designed for all climates ranging from hot and humid Florida to the northern reaches of Alaska. Chapter 1 outlines how investments in energy conservation can be `cost-effective` and actually put money into your pocket each year. Chapter 2 provides an introduction to the issues and factors that determine how much energy your building will consume. Chapter 3 of the Guidebook contains the specific HUD Rehabilitation Energy Guidelines for One-to-Four Family Dwellings. Chapter 4 provides some examples of typical energy conservation measures in various climates. Appendix A is where you will find the climate zone for the appropriate county in your state. In the back of the book, Appendix G contains a blank copy of the One-to-Four Family Cost-Effectiveness Worksheet to copy or tear out for use on your project.

  9. Better Buildings Neighborhood Program Multi-family/ Low Income Peer Exchange Call: Information Technology Tools for Multi-family Building Programs Call Slides and Discussion Summary, March 15, 2012

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    5, 2012 Better Buildings Neighborhood Program Multi-family/ Low Income Peer Exchange Call: Information Technology Tools for Multi-family Building Programs Call Slides and Discussion Summary Agenda * Call Logistics and Attendance  What information technology tools is your program using for project information, marketing, assessment, tracking or evaluation? What do you wish you had? * Program Experience and Lessons:  Heather Larson, StopWaste.org, Energy Upgrade California  Adam Palmer,

  10. Energy Department Awards $92.5 Million to 19 States to Weatherize Homes of Low-Income Families

    Broader source: Energy.gov [DOE]

    WASHINGTON, DC - Secretary of Energy Samuel W. Bodman announced that $92.5 million has been awarded to 19 states to improve the energy efficiency of low-income family homes. The Department of...

  11. The Standard Work Specifications for Single-Family Home Energy Upgrades are now available€ at your fingertips!

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    The Standard Work Specifications for Single-Family Home Energy Upgrades are now availableat your fingertips! The National Renewable Energy Laboratory (NREL) and the U.S. Department of Energy (DOE) are pleased to announce that the Standard Work Specifications for Single-Family Home Energy Upgrades are now available via a user-friendly online tool, with specifications for manufactured housing and multifamily homes to follow. Explore the Standard Work Specifications Online Tool now. Standard work

  12. Philips Light Sources & Electronics is Developing an Efficient, Smaller, Cost-Effective Family of LED Drivers

    Broader source: Energy.gov [DOE]

    With the help of DOE funding, Philips Light Sources & Electronics is developing a new family of LED drivers that are more efficient and cost-effective as well as smaller in size than currently available drivers. The new drivers are switch-mode power supplies that are similar to today's drivers, but with an improved design. In addition, they have a different topology—boost plus LLC—for wattages of 40W and above, but they retain the commonly used flyback topology at lower wattages.

  13. Experimental evidence of negative linear compressibility in the MIL-53 metal–organic framework family

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Serra-Crespo, Pablo; Dikhtiarenko, Alla; Stavitski, Eli; Juan-Alcañiz, Jana; Kapteijn, Freek; Coudert, François-Xavier; Gascon, Jorge

    2014-03-24

    Here we report a series of powder X-ray diffraction experiments performed on the soft porous crystals MIL-53(Al) and NH2-MIL-53(Al) in a diamond anvil cell under different pressurization media. Systematic refinements of the obtained powder patterns demonstrate that these materials expand along a specific direction while undergoing total volume reduction under an increasing hydrostatic pressure. The results confirm for the first time the negative linear compressibility behaviour of this family of materials, recently predicted from quantum chemical calculations.

  14. Experimental Evidence of Negative Linear Compressibility in the MIL-53 Metal-Organic Framework Family

    SciTech Connect (OSTI)

    Serra-Crespo, Pablo; Dikhtiarenko, Alla; Stavitski, Eli; Juan-Alcaniz, Jana; Kapeteijn, Freek; Coudert, Francois-Xavier; Gascon, Jorge

    2014-03-24

    Here, we report a series of powder X-ray diffraction experiments performed on the soft porous crystals MIL-53(Al) and NH2-MIL-53(Al) in a diamond anvil cell under different pressurization media. Systematic refinements of the obtained powder patterns demonstrate that these materials expand along a specific direction while undergoing total volume reduction under an increasing hydrostatic pressure. Our results confirm for the first time the negative linear compressibility behaviour of this family of materials, recently predicted from quantum chemical calculations.

  15. Experimental Evidence of Negative Linear Compressibility in the MIL-53 Metal-organic Framework Family

    SciTech Connect (OSTI)

    Serra-Crespo, Pablo; Dikhtiarenko, Alla; Stavitski, Eli; Juan-Alcaniz, Jana; Kapteijn, Freek; Coudert, Francois-Xavier; Gascon, Jorge

    2014-03-24

    Here we report a series of powder X-ray diffraction experiments performed on the soft porous crystals MIL-53(Al) and NH2-MIL-53(Al) in a diamond anvil cell under different pressurization media. Systematic refinements of the obtained powder patterns demonstrate that these materials expand along a specific direction while undergoing total volume reduction under an increasing hydrostatic pressure. Our results confirm for the first time the negative linear compressibility behaviour of this family of materials, recently predicted from quantum chemical calculations.

  16. Economic analysis of a passive solar multiple-family dwelling for upstate New York

    SciTech Connect (OSTI)

    Laquatra, J. Jr.

    1982-02-01

    The objective of this study was to examine the economic feasibility of passive solar energy as applied to a multiple-family dwelling in three upstate New York cities: Buffalo, Rochester, and Syracuse. Specifically, two passive solar applications - a Trombe wall and a direct-gain system - for a nine-unit structure designed by Total Environmental Action, Inc. were analyzed through the use of a solar economic performance code. City-specific data, including climatological information, building construction costs, utility rates, and property taxes were used, as were various economic parameters to reflect economic conditions in general and specifically those of the solar systems' owners.

  17. An invitation for family fun at PPPL's Open House on June 1 | Princeton

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Plasma Physics Lab An invitation for family fun at PPPL's Open House on June 1 By Jeanne Jackson DeVoe May 13, 2013 Tweet Widget Google Plus One Share on Facebook Adam Cohen, PPPL's deputy director for operations, far left, and engineer James Chrzanowski, far right, both in black, talk to visitors during a tour at PPPL's 2010 Open House. (Photo by Elle Starkman/ PPPL Office of Communications) Adam Cohen, PPPL's deputy director for operations, far left, and engineer James Chrzanowski, far

  18. Structural Characterization of Inhibitors with Selectivity against Members of a Homologous Enzyme Family

    SciTech Connect (OSTI)

    Pavlovsky, Alexander G.; Liu, Xuying; Faehnle, Christopher R.; Potente, Nina; Viola, Ronald E.

    2013-01-31

    The aspartate biosynthetic pathway provides essential metabolites for many important biological functions, including the production of four essential amino acids. As this critical pathway is only present in plants and microbes, any disruptions will be fatal to these organisms. An early pathway enzyme, L-aspartate-{beta}-semialdehyde dehydrogenase, produces a key intermediate at the first branch point of this pathway. Developing potent and selective inhibitors against several orthologs in the L-aspartate-{beta}-semialdehyde dehydrogenase family can serve as lead compounds for antibiotic development. Kinetic studies of two small molecule fragment libraries have identified inhibitors that show good selectivity against L-aspartate-{beta}-semialdehyde dehydrogenases from two different bacterial species, Streptococcus pneumoniae and Vibrio cholerae, despite the presence of an identical constellation of active site amino acids in this homologous enzyme family. Structural characterization of enzyme-inhibitor complexes have elucidated different modes of binding between these structurally related enzymes. This information provides the basis for a structure-guided approach to the development of more potent and more selective inhibitors.

  19. The actin family protein ARP6 contributes to the structure and the function of the nucleolus

    SciTech Connect (OSTI)

    Kitamura, Hiroshi; Matsumori, Haruka; Kalendova, Alzbeta; Hozak, Pavel; Goldberg, Ilya G.; Nakao, Mitsuyoshi; Saitoh, Noriko; Harata, Masahiko

    2015-08-21

    The actin family members, consisting of actin and actin-related proteins (ARPs), are essential components of chromatin remodeling complexes. ARP6, one of the nuclear ARPs, is part of the Snf-2-related CREB-binding protein activator protein (SRCAP) chromatin remodeling complex, which promotes the deposition of the histone variant H2A.Z into the chromatin. In this study, we showed that ARP6 influences the structure and the function of the nucleolus. ARP6 is localized in the central region of the nucleolus, and its knockdown induced a morphological change in the nucleolus. We also found that in the presence of high concentrations of glucose ARP6 contributed to the maintenance of active ribosomal DNA (rDNA) transcription by placing H2A.Z into the chromatin. In contrast, under starvation, ARP6 was required for cell survival through the repression of rDNA transcription independently of H2A.Z. These findings reveal novel pleiotropic roles for the actin family in nuclear organization and metabolic homeostasis. - Highlights: • ARP6, an actin related protein, is important for nucleolar function and structure. • A population of ARP6 is localized in the center of nucleolus. • Depletion of ARP6 resulted in aberrant shape of the nucleolus. • ARP6 maintains the active rDNA transcription under high glucose. • ARP6 is required for the repression of rDNA transcription under starvation.

  20. Biochemical and structural characterizations of two Dictyostelium cellobiohydrolases from the amoebozoa kingdom reveal a high level of conservation between distant phylogenetic trees of life

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Hobdey, Sarah E.; Knott, Brandon C.; Momeni, Majid Haddad; Taylor, II, Larry E.; Borisova, Anna S.; Podkaminer, Kara K.; VanderWall, Todd A.; Himmel, Michael E.; Decker, Stephen R.; Beckham, Gregg T.; et al

    2016-04-01

    Glycoside hydrolase family 7 (GH7) cellobiohydrolases (CBHs) are enzymes often employed in plant cell wall degradation across eukaryotic kingdoms of life, as they provide significant hydrolytic potential in cellulose turnover. To date, many fungal GH7 CBHs have been examined, yet many questions regarding structure-activity relationships in these important natural and commercial enzymes remain. Here, we present the crystal structures and a biochemical analysis of two GH7 CBHs from social amoeba: Dictyostelium discoideum Cel7A (DdiCel7A) and Dictyostelium purpureum Cel7A (DpuCel7A). DdiCel7A and DpuCel7A natively consist of a catalytic domain and do not exhibit a carbohydrate-binding module (CBM). The structures of DdiCel7Amore » and DpuCel7A, resolved to 2.1 Å and 2.7 Å, respectively, are homologous to those of other GH7 CBHs with an enclosed active-site tunnel. Two primary differences between the Dictyostelium CBHs and the archetypal model GH7 CBH, Trichoderma reesei Cel7A (TreCel7A), occur near the hydrolytic active site and the product-binding sites. To compare the activities of these enzymes with the activity of TreCel7A, the family 1 TreCel7A CBM and linker were added to the C terminus of each of the Dictyostelium enzymes, creating DdiCel7ACBM and DpuCel7ACBM, which were recombinantly expressed in T. reesei. DdiCel7ACBM and DpuCel7ACBM hydrolyzed Avicel, pretreated corn stover, and phosphoric acid-swollen cellulose as efficiently as TreCel7A when hydrolysis was compared at their temperature optima. The Ki of cellobiose was significantly higher for DdiCel7ACBM and DpuCel7ACBM than for TreCel7A: 205, 130, and 29 μM, respectively. Finally, taken together, the present study highlights the remarkable degree of conservation of the activity of these key natural and industrial enzymes across quite distant phylogenetic trees of life.« less

  1. Biochemical and Structural Characterization of Two Dictyostelium Cellobiohydrolases from the Amoebozoa Kingdom Reveal a High Level of Conservation Between Distant Phylogenetic Trees of Life

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Hobdey, Sarah E.; Knott, Brandon C.; Momeni, Majid Haddad; Taylor, II, Larry E.; Borisova, Anna S.; Podkaminer, Kara K.; VanderWall, Todd A.; Himmel, Michael E.; Decker, Stephen R.; Beckham, Gregg T.; et al

    2016-06-01

    Glycoside Hydrolase Family 7 (GH7) cellobiohydrolases (CBHs) are commonly employed enzymes in plant cell wall degradation across eukaryotic kingdoms of life, as they provide significant hydrolytic potential in cellulose turnover. To date, many fungal GH7 CBHs have been examined, yet many questions remain regarding structure-activity relationships in these important natural and commercial enzymes. Here, we present crystal structures and biochemical analysis of two GH7 CBHs from social amoeba: Dictyostelium discoideum and Dictyostelium purpureum (DdiCel7A and DpuCel7A, respectively). DdiCel7A and DpuCel7A natively consist of a catalytic domain and do not exhibit a carbohydrate-binding module (CBM). The structures, resolved to 2.1 Amore » (DdiCel7A), and 2.7 A (DpuCel7A), are homologous to other GH7 CBHs with an enclosed active site tunnel. Two primary differences between the Dictyostelium CBHs and the archetypal model GH7 CBH from Trichoderma reesei Cel7A (TreCel7A) occur near the hydrolytic active site and the product binding sites. To compare the activity of these enzymes with TreCel7A, the Family 1 TreCel7A CBM and linker was added to the C-terminus of the Dictyostelium enzymes, DdiCel7ACBM and DpuCel7ACBM, which were recombinantly expressed in T. reesei. DdiCel7ACBM and DpuCel7ACBM hydrolyze Avicel, pretreated corn stover, and phosphoric acid swollen cellulose as efficiently as TreCel7A when compared at their temperature optima. The Ki of cellobiose is significantly higher for DdiCel7ACBM and DpuCel7ACBM than for TreCel7A: 205, 130, and 29 uM, respectively. Taken together, the present study highlights the remarkable conservation in the activity of these key natural and industrial enzymes across quite distant phylogenetic trees of life.« less

  2. Support and utilization of the LSI-11 processor family at SLAC

    SciTech Connect (OSTI)

    Kieffer, J.; Logg, C.A.; Farwell, D.E.

    1981-01-01

    Microcomputer systems based on the DEC LSI-11 processor family have been in use at SLAC for five years. They are used for a wide variety of applications. The support of these systems is divided into three general areas: engineering, maintenance, and software. Engineering specifies the system to match user requirements. SLAC has been able to design one general purpose system which can be tailored to fit many specific requirements. Maintenance provides system and component diagnostic services and repair. Software support includes software consulting services, assistance in systems design, and the development and support of special purpose operating systems and programs. These support functions are handled as subtasks by three teams in the SLAC Electronics Instrumentation Group. Each of these teams utilizes several LSI-11 systems in the performance of its primary tasks. They work closely together to jointly provide overall support for the larger SLAC community.

  3. Installation guidelines for Solar Heating System, single-family residence at New Castle, Pennsylvania

    SciTech Connect (OSTI)

    Not Available

    1980-01-01

    The Solar Heating System installer guidelines are provided for each subsystem and includes testing and filling the system. This single-family residential heating system is a solar-assisted, hydronic-to-warm-air system with solar-assisted domestic water heating. It is composed of the following major components: liquid cooled flat plate collectors; water storage tank; passive solar-fired domestic water preheater; electric hot water heater; heat pump with electric backup; solar hot water coil unit; tube-and-shell heat exchanger, three pumps, and associated pipes and valving in an energy transport module; control system; and air-cooled heat purge unit. Information is also provided on the operating procedures, controls, caution requirements, and routine and schedule maintenance. Information consists of written procedures, schematics, detail drawings, pictures and manufacturer's component data.

  4. Adverse reproductive outcomes in families of atomic veterans: The feasibility of epidemiologic studies

    SciTech Connect (OSTI)

    1995-12-31

    This is an outstanding report from a distinguished academy committee, which in 71 pages of text provides the scientific basis for the carefully crafted 8-page executive summary. The principles and issues of the required epidemiological study are presented calmly and concisely, as are the ensuing short chapters on radiation biology, genetics and risk estimation, and all other adverse reproductive outcomes. The committee was mandated by Congress to determine the feasibility, cost and duration of a study on adverse reproductive outcomes in families of atomic veteran. The committee found that a scientifically adequate and epidemiologically valid study could not be mounted and the cost would be tens of millions of dollars lasting a decade. The Committee presents a number of well-discussed approaches in support of their position.

  5. Single Dirac Cone Topological Surface State and Unusual Thermoelectric Property of Compounds from a New Topological Insulator Family

    SciTech Connect (OSTI)

    Chen, Y

    2011-08-18

    Angle resolved photoemission spectroscopy (ARPES) study on TlBiTe2 and TlBiSe2 from a Thallium-based III-V-VI2 ternary chalcogenides family revealed a single surface Dirac cone at the center of the Brillouin zone for both compounds. For TlBiSe{sub 2}, the large bulk gap ({approx} 200meV) makes it a topological insulator with better mechanical properties than the previous binary 3D topological insualtor family. For TlBiTe{sub 2}, the observed negative bulk gap indicates it as a semi-metal, rather than a narrow gap semi-conductor as conventionally believed; this semi-metality naturally explains its mysteriously small thermoelectric figure of merit comparing to other compounds in the family. Finally, the unique band structures of TlBiTe{sub 2} also suggests it as a candidate for topological superconductors.

  6. Pathways and barriers to genetic testing and screening: Molecular genetics meets the high-risk family. Final report

    SciTech Connect (OSTI)

    Duster, T.

    1998-11-01

    The proliferation of genetic screening and testing is requiring increasing numbers of Americans to integrate genetic knowledge and interventions into their family life and personal experience. This study examines the social processes that occur as families at risk for two of the most common autosomal recessive diseases, sickle cell disease (SC) and cystic fibrosis (CF), encounter genetic testing. Each of these diseases is found primarily in a different ethnic/racial group (CF in Americans of North European descent and SC in Americans of West African descent). This has permitted them to have a certain additional lens on the role of culture in integrating genetic testing into family life and reproductive planning. A third type of genetic disorder, the thalassemias was added to the sample in order to extent the comparative frame and to include other ethnic and racial groups.

  7. Novel inherited mutations and variable expressivity of BRCA1 alleles, including the founder mutation 185delAG in Ashkenazi Jewish families

    SciTech Connect (OSTI)

    Friedman, L.S.; Szabo, C.I.; Ostermeyer, E.A.

    1995-12-01

    Thirty-seven families with four or more cases of breast cancer or breast and ovarian cancer were analyzed for mutations in BRCA1. Twelve different germ-line mutations, four novel and eight previously observed, were detected in 16 families. Five families of Ashkenazi Jewish descent carried the 185delAG mutation and shared the same haplotype at eight polymorphic markers spanning {approximately}850 kb at BRCA1. Expressivity of 185delAG in these families varied, from early-onset bilateral breast cancer and ovarian cancer to late-onset breast cancer without ovarian cancer. Mutation 4184delTCAA occurred independently in two families. In one family, penetrance was complete, with females developing early-onset breast cancer or ovarian cancer and the male carrier developing prostatic cancer, whereas, in the other family, penetrance was incomplete and only breast cancer occurred, diagnosed at ages 38-81 years. Two novel nonsense mutations led to the loss of mutant BRCA1 transcript in families with 10 and 6 cases of early-onset breast cancer and ovarian cancer. A 665-nt segment of the BRCA1 3{prime}-UTR and 1.3 kb of genomic sequence including the putative promoter region were invariant by single-strand conformation analysis in 13 families without coding-sequence mutations. Overall in our series, BRCA1 mutations have been detected in 26 families: 16 with positive BRCA1 lod scores, 7 with negative lod scores (reflecting multiple sporadic breast cancers), and 3 not tested for linkage. Three other families have positive lod scores for linkage to BRCA2, but 13 families without detected BRCA1 mutations have negative lod scores for both BRCA1 and BRCA2. 57 refs., 5 figs., 3 tabs.

  8. Subunit composition and glycosidic activities of the cellulase complex from Clostridium thermocellum JW20

    SciTech Connect (OSTI)

    Kohring, S.; Mayer, F. ); Wiegel, J. )

    1990-12-01

    The subunit composition of the extracellular complex from Clostridium thermocellum was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Twenty-six bands, representing proteins with apparent molecular sizes ranging from 37,500 to 185,000 Da, could be detected by silver staining. Endoglucanase activity was exhibited in 15 of 26 bands and 13 showed xylanase activity. In 8 of the 26 bands, both activities could be found. As minor activities, {beta}-glucosidase, {beta}-xylosidase, {beta}-galactosidase, and {beta}-mannosidase activities could be demonstrated in the cellulase complex. Upon measuring the release of para-nitrophenol (PNP) from PNP-cellobioside and determining the amount of glucose formed, the presence of exoglucanase activity was indicated. Upon glycoprotein staining of SDS-polyacrylamide gels, 14 of the 26 bands reacted positive, indicating the glycoprotein nature of the respective proteins. Four proteins (apparent molecular sizes, 58,000, 72,500, 94,000, and 110,000 Da) could be enriched from the originally bound cellulase complex by preparative SDS-PAGE. The two smaller proteins exhibited xylanase activity, whereas the 94,000-Da protein had endo- and exoglucanase activity, and the 110,000-Da protein degraded PNP-pyranosides.

  9. Measured energy savings and economics of retrofitting existing single- family homes: An update of the BECA-B database

    SciTech Connect (OSTI)

    Cohen, S.D.; Goldman, C.A.; Harris, J.P.

    1991-02-01

    These appendices are the companion volume to report number LBL--28147 Vol.1, with the same title. The summary data tables include physical characteristics, energy consumption, savings, and the retrofit measures installed and their costs for each retrofit project. Each existing single family residential building'' retrofit project in the BECA-B database is described. 99 refs. (BM)

  10. Binding Preferences, Surface Attachment, Diffusivity, and Orientation of a Family 1 Carbohydrate-Binding Module on Cellulose

    SciTech Connect (OSTI)

    Nimlos, M. R.; Beckham, G. T.; Matthews, J. F.; Bu, L.; Himmel, M. E.; Crowley, M. F.

    2012-06-08

    Cellulase enzymes often contain carbohydrate-binding modules (CBMs) for binding to cellulose. The mechanisms by which CBMs recognize specific surfaces of cellulose and aid in deconstruction are essential to understand cellulase action. The Family 1 CBM from the Trichoderma reesei Family 7 cellobiohydrolase, Cel7A, is known to selectively bind to hydrophobic surfaces of native cellulose. It is most commonly suggested that three aromatic residues identify the planar binding face of this CBM, but several recent studies have challenged this hypothesis. Here, we use molecular simulation to study the CBM binding orientation and affinity on hydrophilic and hydrophobic cellulose surfaces. Roughly 43 {mu}s of molecular dynamics simulations were conducted, which enables statistically significant observations. We quantify the fractions of the CBMs that detach from crystal surfaces or diffuse to other surfaces, the diffusivity along the hydrophobic surface, and the overall orientation of the CBM on both hydrophobic and hydrophilic faces. The simulations demonstrate that there is a thermodynamic driving force for the Cel7A CBM to bind preferentially to the hydrophobic surface of cellulose relative to hydrophilic surfaces. In addition, the simulations demonstrate that the CBM can diffuse from hydrophilic surfaces to the hydrophobic surface, whereas the reverse transition is not observed. Lastly, our simulations suggest that the flat faces of Family 1 CBMs are the preferred binding surfaces. These results enhance our understanding of how Family 1 CBMs interact with and recognize specific cellulose surfaces and provide insights into the initial events of cellulase adsorption and diffusion on cellulose.