Expression of a heat-stable NADPH-dependent alcohol dehydrogenase from Thermoanaerobacter pseudethanolicus 39E in Clostridium thermocellum 1313 results in increased hydroxymethylfurfural resistance

Kim, Sun -Ki; Groom, Joseph; Chung, Daehwan; Elkins, James; Westpheling, Janet

doi:10.1186/s13068-017-0750-z

Title: Expression of a heat-stable NADPH-dependent alcohol dehydrogenase from Thermoanaerobacter pseudethanolicus 39E in Clostridium thermocellum 1313 results in increased hydroxymethylfurfural resistance

Journal Article · 15 March 2017 · Biotechnology for Biofuels

DOI: https://doi.org/10.1186/s13068-017-0750-z · OSTI ID:1350020

Kim, Sun -Ki ^[1]; Groom, Joseph ^[1]; Chung, Daehwan ^[2]; Elkins, James ^[3]; Westpheling, Janet ^[1]

Univ. of Georgia, Athens, GA (United States); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
National Renewable Energy Lab. (NREL), Golden, CO (United States); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)

Resistance to deconstruction is a major limitation to the use of lignocellulosic biomass as a substrate for the production of fuels and chemicals. Consolidated bioprocessing (CBP), the use of microbes for the simultaneous hydrolysis of lignocellulose into soluble sugars and fermentation of the resulting sugars to products of interest, is a potential solution to this obstacle. The pretreatment of plant biomass, however, releases compounds that are inhibitory to the growth of microbes used for CBP. Heterologous expression of the Thermoanaerobacter pseudethanolicus 39E bdhA gene, that encodes an alcohol dehydrogenase, in Clostridium thermocellum significantly increased resistance to furan derivatives at concentrations found in acid-pretreated biomass. The mechanism of detoxification of hydroxymethylfurfural was shown to be primarily reduction using NADPH as the cofactor. In addition, we report the construction of new expression vectors for homologous and heterologous expression in C. thermocellum. These vectors use regulatory signals from both C. bescii (the S-layer promoter) and C. thermocellum (the enolase promoter) shown to efficiently drive expression of the BdhA enzyme. Toxic compounds present in lignocellulose hydrolysates that inhibit cell growth and product formation are obstacles to the commercialization of fuels and chemicals from biomass. Lastly, expression of genes that reduce the effect of these inhibitors, such as furan derivatives, will serve to enable commercial processes using plant biomass for the production of fuels and chemicals.

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Research Organization:: NREL (National Renewable Energy Laboratory (NREL), Golden, CO (United States))

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)

Grant/Contract Number:: AC36-08GO28308

OSTI ID:: 1350020

Report Number(s):: NREL/JA--2700-68259

Journal Information:: Biotechnology for Biofuels, Journal Name: Biotechnology for Biofuels Journal Issue: 1 Vol. 10; ISSN 1754-6834

Publisher:: BioMed CentralCopyright Statement

Country of Publication:: United States

Language:: English

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A 5-hydroxymethyl furfural reducing enzyme encoded by theSaccharomyces cerevisiae ADH6 gene conveys HMF tolerance Petersson, Anneli; Almeida, João R. M.; Modig, Tobias Yeast, Vol. 23, Issue 6, p. 455-464 https://doi.org/10.1002/yea.1370	journal	January 2006
NADH- vs NADPH-coupled reduction of 5-hydroxymethyl furfural (HMF) and its implications on product distribution in Saccharomyces cerevisiae Almeida, João R. M.; Röder, Anja; Modig, Tobias Applied Microbiology and Biotechnology, Vol. 78, Issue 6 https://doi.org/10.1007/s00253-008-1364-y	journal	March 2008
Multiple gene-mediated NAD(P)H-dependent aldehyde reduction is a mechanism of in situ detoxification of furfural and 5-hydroxymethylfurfural by Saccharomyces cerevisiae Lewis Liu, Z.; Moon, Jaewoong; Andersh, Brad J. Applied Microbiology and Biotechnology, Vol. 81, Issue 4, p. 743-753 https://doi.org/10.1007/s00253-008-1702-0	journal	September 2008
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Expression of aldehyde dehydrogenase 6 reduces inhibitory effect of furan derivatives on cell growth and ethanol production in Saccharomyces cerevisiae Park, Seong-Eon; Koo, Hyun Min; Park, Young Kyoung Bioresource Technology, Vol. 102, Issue 10, p. 6033-6038 https://doi.org/10.1016/j.biortech.2011.02.101	journal	May 2011
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Redirecting carbon flux through exogenous pyruvate kinase to achieve high ethanol yields in Clostridium thermocellum Deng, Yu; Olson, Daniel G.; Zhou, Jilai Metabolic Engineering, Vol. 15, p. 151-158 https://doi.org/10.1016/j.ymben.2012.11.006	journal	January 2013
Enhanced tolerance of Saccharomyces cerevisiae to multiple lignocellulose-derived inhibitors through modulation of spermidine contents Kim, Sun-Ki; Jin, Yong-Su; Choi, In-Geol Metabolic Engineering, Vol. 29 https://doi.org/10.1016/j.ymben.2015.02.004	journal	May 2015
Engineering electron metabolism to increase ethanol production in Clostridium thermocellum Lo, Jonathan; Olson, Daniel G.; Murphy, Sean Jean-Loup Metabolic Engineering, Vol. 39 https://doi.org/10.1016/j.ymben.2016.10.018	journal	January 2017
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Insights into plant biomass conversion from the genome of the anaerobic thermophilic bacterium Caldicellulosiruptor bescii DSM 6725 Dam, Phuongan; Kataeva, Irina; Yang, Sung-Jae Nucleic Acids Research, Vol. 39, Issue 8 https://doi.org/10.1093/nar/gkq1281	journal	January 2011
Efficient Degradation of Lignocellulosic Plant Biomass, without Pretreatment, by the Thermophilic Anaerobe "Anaerocellum thermophilum" DSM 6725 Yang, Sung-Jae; Kataeva, Irina; Hamilton-Brehm, Scott D. Applied and Environmental Microbiology, Vol. 75, Issue 14, p. 4762-4769 https://doi.org/10.1128/aem.00236-09	journal	May 2009
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A rapid chemical method for lysing Arabidopsis cells for protein analysis Tsugama, Daisuke; Liu, Shenkui; Takano, Tetsuo Plant Methods, Vol. 7, Issue 1 https://doi.org/10.1186/1746-4811-7-22	journal	January 2011
The influence of HMF and furfural on redox-balance and energy-state of xylose-utilizing Saccharomyces cerevisiae Ask, Magnus; Bettiga, Maurizio; Mapelli, Valeria Biotechnology for Biofuels, Vol. 6, Issue 1 https://doi.org/10.1186/1754-6834-6-22	journal	January 2013
Elimination of hydrogenase active site assembly blocks H2 production and increases ethanol yield in Clostridium thermocellum Biswas, Ranjita; Zheng, Tianyong; Olson, Daniel G. Biotechnology for Biofuels, Vol. 8, Issue 1 https://doi.org/10.1186/s13068-015-0204-4	journal	January 2015
Expression of a heat-stable NADPH-dependent alcohol dehydrogenase in Caldicellulosiruptor bescii results in furan aldehyde detoxification Chung, Daehwan; Verbeke, Tobin J.; Cross, Karissa L. Biotechnology for Biofuels, Vol. 8, Issue 1 https://doi.org/10.1186/s13068-015-0287-y	journal	July 2015
The emergence of Clostridium thermocellum as a high utility candidate for consolidated bioprocessing applications Akinosho, Hannah; Yee, Kelsey; Close, Dan Frontiers in Chemistry, Vol. 2 https://doi.org/10.3389/fchem.2014.00066	journal	August 2014
Increased tolerance and conversion of inhibitors in lignocellulosic hydrolysates by Saccharomyces cerevisiae Almeida, Joao; Modig, Tobias; Petersson, Anneli Journal of Chemical Technology & Biotechnology, Vol. 82, Issue 4, p. 340-349 https://doi.org/10.1002/jctb.1676	journal	January 2007
A 5-hydroxymethyl furfural reducing enzyme encoded by theSaccharomyces cerevisiae ADH6 gene conveys HMF tolerance Petersson, Anneli; Almeida, João R. M.; Modig, Tobias Yeast, Vol. 23, Issue 6, p. 455-464 https://doi.org/10.1002/yea.1370	journal	January 2006
NADH- vs NADPH-coupled reduction of 5-hydroxymethyl furfural (HMF) and its implications on product distribution in Saccharomyces cerevisiae Almeida, João R. M.; Röder, Anja; Modig, Tobias Applied Microbiology and Biotechnology, Vol. 78, Issue 6 https://doi.org/10.1007/s00253-008-1364-y	journal	March 2008
Multiple gene-mediated NAD(P)H-dependent aldehyde reduction is a mechanism of in situ detoxification of furfural and 5-hydroxymethylfurfural by Saccharomyces cerevisiae Lewis Liu, Z.; Moon, Jaewoong; Andersh, Brad J. Applied Microbiology and Biotechnology, Vol. 81, Issue 4, p. 743-753 https://doi.org/10.1007/s00253-008-1702-0	journal	September 2008
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Effect of dilute acid pretreatment of rice straw on structural properties and enzymatic hydrolysis Hsu, Teng-Chieh; Guo, Gia-Luen; Chen, Wen-Hua Bioresource Technology, Vol. 101, Issue 13 https://doi.org/10.1016/j.biortech.2009.10.009	journal	July 2010
Expression of aldehyde dehydrogenase 6 reduces inhibitory effect of furan derivatives on cell growth and ethanol production in Saccharomyces cerevisiae Park, Seong-Eon; Koo, Hyun Min; Park, Young Kyoung Bioresource Technology, Vol. 102, Issue 10, p. 6033-6038 https://doi.org/10.1016/j.biortech.2011.02.101	journal	May 2011
Recent progress in consolidated bioprocessing Olson, Daniel G.; McBride, John E.; Joe Shaw, A. Current Opinion in Biotechnology, Vol. 23, Issue 3, p. 396-405 https://doi.org/10.1016/j.copbio.2011.11.026	journal	June 2012
Development of a high-throughput method to evaluate the impact of inhibitory compounds from lignocellulosic hydrolysates on the growth of Zymomonas mobilis Franden, Mary Ann; Pienkos, Philip T.; Zhang, Min Journal of Biotechnology, Vol. 144, Issue 4 https://doi.org/10.1016/j.jbiotec.2009.08.006	journal	December 2009
Identifying promoters for gene expression in Clostridium thermocellum Olson, Daniel G.; Maloney, Marybeth; Lanahan, Anthony A. Metabolic Engineering Communications, Vol. 2 https://doi.org/10.1016/j.meteno.2015.03.002	journal	December 2015
Promiscuous plasmid replication in thermophiles: Use of a novel hyperthermophilic replicon for genetic manipulation of Clostridium thermocellum at its optimum growth temperature Groom, Joseph; Chung, Daehwan; Olson, Daniel G. Metabolic Engineering Communications, Vol. 3 https://doi.org/10.1016/j.meteno.2016.01.004	journal	December 2016
Cloning, sequencing, and sequence analysis of two novel plasmids from the thermophilic anaerobic bacterium Anaerocellum thermophilum Clausen, Anders; Mikkelsen, Marie Just; Schröder, Imke Plasmid, Vol. 52, Issue 2 https://doi.org/10.1016/j.plasmid.2004.06.001	journal	September 2004
Redirecting carbon flux through exogenous pyruvate kinase to achieve high ethanol yields in Clostridium thermocellum Deng, Yu; Olson, Daniel G.; Zhou, Jilai Metabolic Engineering, Vol. 15, p. 151-158 https://doi.org/10.1016/j.ymben.2012.11.006	journal	January 2013
Enhanced tolerance of Saccharomyces cerevisiae to multiple lignocellulose-derived inhibitors through modulation of spermidine contents Kim, Sun-Ki; Jin, Yong-Su; Choi, In-Geol Metabolic Engineering, Vol. 29 https://doi.org/10.1016/j.ymben.2015.02.004	journal	May 2015
Elimination of metabolic pathways to all traditional fermentation products increases ethanol yields in Clostridium thermocellum Papanek, Beth; Biswas, Ranjita; Rydzak, Thomas Metabolic Engineering, Vol. 32 https://doi.org/10.1016/j.ymben.2015.09.002	journal	November 2015
Engineering electron metabolism to increase ethanol production in Clostridium thermocellum Lo, Jonathan; Olson, Daniel G.; Murphy, Sean Jean-Loup Metabolic Engineering, Vol. 39 https://doi.org/10.1016/j.ymben.2016.10.018	journal	January 2017
Mutant alcohol dehydrogenase leads to improved ethanol tolerance in Clostridium thermocellum Brown, S. D.; Guss, A. M.; Karpinets, T. V. Proceedings of the National Academy of Sciences, Vol. 108, Issue 33 https://doi.org/10.1073/pnas.1102444108	journal	August 2011
Engineering furfural tolerance in Escherichia coli improves the fermentation of lignocellulosic sugars into renewable chemicals Wang, X.; Yomano, L. P.; Lee, J. Y. Proceedings of the National Academy of Sciences, Vol. 110, Issue 10, p. 4021-4026 https://doi.org/10.1073/pnas.1217958110	journal	February 2013
Direct conversion of plant biomass to ethanol by engineered Caldicellulosiruptor bescii Chung, D.; Cha, M.; Guss, A. M. Proceedings of the National Academy of Sciences, Vol. 111, Issue 24, p. 8931-8936 https://doi.org/10.1073/pnas.1402210111	journal	June 2014
Insights into plant biomass conversion from the genome of the anaerobic thermophilic bacterium Caldicellulosiruptor bescii DSM 6725 Dam, Phuongan; Kataeva, Irina; Yang, Sung-Jae Nucleic Acids Research, Vol. 39, Issue 8 https://doi.org/10.1093/nar/gkq1281	journal	January 2011
Efficient Degradation of Lignocellulosic Plant Biomass, without Pretreatment, by the Thermophilic Anaerobe "Anaerocellum thermophilum" DSM 6725 Yang, Sung-Jae; Kataeva, Irina; Hamilton-Brehm, Scott D. Applied and Environmental Microbiology, Vol. 75, Issue 14, p. 4762-4769 https://doi.org/10.1128/AEM.00236-09	journal	May 2009
Development of pyrF-Based Genetic System for Targeted Gene Deletion in Clostridium thermocellum and Creation of a pta Mutant Tripathi, S. A.; Olson, D. G.; Argyros, D. A. Applied and Environmental Microbiology, Vol. 76, Issue 19, p. 6591-6599 https://doi.org/10.1128/AEM.01484-10	journal	August 2010
A rapid chemical method for lysing Arabidopsis cells for protein analysis Tsugama, Daisuke; Liu, Shenkui; Takano, Tetsuo Plant Methods, Vol. 7, Issue 1 https://doi.org/10.1186/1746-4811-7-22	journal	January 2011
The influence of HMF and furfural on redox-balance and energy-state of xylose-utilizing Saccharomyces cerevisiae Ask, Magnus; Bettiga, Maurizio; Mapelli, Valeria Biotechnology for Biofuels, Vol. 6, Issue 1 https://doi.org/10.1186/1754-6834-6-22	journal	January 2013
A comparative multidimensional LC-MS proteomic analysis reveals mechanisms for furan aldehyde detoxification in Thermoanaerobacter pseudethanolicus 39E Clarkson, Sonya M.; Hamilton-Brehm, Scott D.; Giannone, Richard J. Biotechnology for Biofuels, Vol. 7, Issue 1 https://doi.org/10.1186/s13068-014-0165-z	journal	December 2014
Elimination of hydrogenase active site assembly blocks H2 production and increases ethanol yield in Clostridium thermocellum Biswas, Ranjita; Zheng, Tianyong; Olson, Daniel G. Biotechnology for Biofuels, Vol. 8, Issue 1 https://doi.org/10.1186/s13068-015-0204-4	journal	January 2015
Expression of a heat-stable NADPH-dependent alcohol dehydrogenase in Caldicellulosiruptor bescii results in furan aldehyde detoxification Chung, Daehwan; Verbeke, Tobin J.; Cross, Karissa L. Biotechnology for Biofuels, Vol. 8, Issue 1 https://doi.org/10.1186/s13068-015-0287-y	journal	July 2015
Construction of a Stable Replicating Shuttle Vector for Caldicellulosiruptor Species: Use for Extending Genetic Methodologies to Other Members of This Genus Chung, Daehwan; Cha, Minseok; Farkas, Joel PLoS ONE, Vol. 8, Issue 5, Article No. e62881 https://doi.org/10.1371/journal.pone.0062881	journal	May 2013
Homologous Expression of the Caldicellulosiruptor bescii CelA Reveals that the Extracellular Protein Is Glycosylated Chung, Daehwan; Young, Jenna; Bomble, Yannick J. PLOS ONE, Vol. 10, Issue 3 https://doi.org/10.1371/journal.pone.0119508	journal	March 2015
The emergence of Clostridium thermocellum as a high utility candidate for consolidated bioprocessing applications Akinosho, Hannah; Yee, Kelsey; Close, Dan Frontiers in Chemistry, Vol. 2 https://doi.org/10.3389/fchem.2014.00066	journal	August 2014

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Related Subjects

09 BIOMASS FUELS
5-hydroxymethyl-2-furfural
Clostridium thermocellum
butanol dehydrogenase
consolidated bioprocessing
furfural

Expression of a heat-stable NADPH-dependent alcohol dehydrogenase from Thermoanaerobacter pseudethanolicus 39E in Clostridium thermocellum 1313 results in increased hydroxymethylfurfural resistance Kim, Sun-Ki; Groom, Joseph; Chung, Daehwan https://doi.org/10.6084/m9.figshare.c.3718192 The current record is supplemented by this dataset	dataset	March 2017
Expression of a heat-stable NADPH-dependent alcohol dehydrogenase from Thermoanaerobacter pseudethanolicus 39E in Clostridium thermocellum 1313 results in increased hydroxymethylfurfural resistance [Supplemental Data] Kim, Sun-Ki; Groom, Joseph; Chung, Daehwan https://doi.org/10.6084/m9.figshare.c.3718192.v1 The current record is supplemented by this dataset	dataset	March 2017
MOESM1 of Expression of a heat-stable NADPH-dependent alcohol dehydrogenase from Thermoanaerobacter pseudethanolicus 39E in Clostridium thermocellum 1313 results in increased hydroxymethylfurfural resistance Kim, Sun-Ki; Groom, Joseph; Chung, Daehwan https://doi.org/10.6084/m9.figshare.c.3718192_d1 The current record is supplemented by this text	text	January 2017
MOESM1 of Expression of a heat-stable NADPH-dependent alcohol dehydrogenase from Thermoanaerobacter pseudethanolicus 39E in Clostridium thermocellum 1313 results in increased hydroxymethylfurfural resistance Kim, Sun-Ki; Groom, Joseph; Chung, Daehwan https://doi.org/10.6084/m9.figshare.c.3718192_d1.v1 The current record is supplemented by this dataset	dataset	March 2017