The ethanol pathway from Thermoanaerobacterium saccharolyticum improves ethanol production in Clostridium thermocellum

Hon, Shuen; Olson, Daniel G.; Holwerda, Evert K.; Lanahan, Anthony A.; Murphy, Sean J. L.; Maloney, Marybeth I.; Zheng, Tianyong; Papanek, Beth; Guss, Adam M.; Lynd, Lee R.

doi:10.1016/j.ymben.2017.06.011

Title: The ethanol pathway from Thermoanaerobacterium saccharolyticum improves ethanol production in Clostridium thermocellum

Journal Article · Tue Jun 27 00:00:00 EDT 2017 · Metabolic Engineering

DOI:https://doi.org/10.1016/j.ymben.2017.06.011· OSTI ID:1394382

^[1]; Olson, Daniel G. ^[1]; Holwerda, Evert K. ^[1]; Lanahan, Anthony A. ^[1]; Murphy, Sean J. L. ^[1]; Maloney, Marybeth I. ^[1]; Zheng, Tianyong ^[1]; Papanek, Beth ^[2]; Guss, Adam M. ^[2]; Lynd, Lee R. ^[1]

Dartmouth College, Hanover, NH (United States); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)

Clostridium thermocellum ferments cellulose, is a promising candidate for ethanol production from cellulosic biomass, and has been the focus of studies aimed at improving ethanol yield. Thermoanaerobacterium saccharolyticum ferments hemicellulose, but not cellulose, and has been engineered to produce ethanol at high yield and titer. Recent research has led to the identification of four genes in T. saccharolyticum involved in ethanol production: adhE, nfnA, nfnB and adhA. We introduced these genes into C. thermocellum and observed significant improvements to ethanol yield, titer, and productivity. The four genes alone, however, were insufficient to achieve in C. thermocellum the ethanol yields and titers observed in engineered T. saccharolyticum strains, even when combined with gene deletions targeting hydrogen production. Here, this suggests that other parts of T. saccharolyticum metabolism may also be necessary to reproduce the high ethanol yield and titer phenotype in C. thermocellum.

View Accepted Manuscript (DOE)

View Accepted Manuscript (Publisher)

Cite

Export

Save

Research Organization:: Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States); Dartmouth College, Hanover, NH (United States)

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

Grant/Contract Number:: AC05-00OR22725; AC02-05CH11231

OSTI ID:: 1394382

Alternate ID(s):: OSTI ID: 1367525; OSTI ID: 1550373

Journal Information:: Metabolic Engineering, Vol. 42, Issue C; ISSN 1096-7176

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

Citation Metrics:

Cited by: 41 works

Citation information provided by
Web of Science

References (34)

High Ethanol Titers from Cellulose by Using Metabolically Engineered Thermophilic, Anaerobic Microbes Argyros, D. Aaron; Tripathi, Shital A.; Barrett, Trisha F. Applied and Environmental Microbiology, Vol. 77, Issue 23, p. 8288-8294 https://doi.org/10.1128/AEM.00646-11	journal	September 2011
Increase in Ethanol Yield via Elimination of Lactate Production in an Ethanol-Tolerant Mutant of Clostridium thermocellum Biswas, Ranjita; Prabhu, Sandeep; Lynd, Lee R. PLoS ONE, Vol. 9, Issue 2 https://doi.org/10.1371/journal.pone.0086389	journal	February 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
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
Bacteria engineered for fuel ethanol production: current status Dien, B. S.; Cotta, M. A.; Jeffries, T. W. Applied Microbiology and Biotechnology, Vol. 63, Issue 3, p. 258-266 https://doi.org/10.1007/s00253-003-1444-y	journal	December 2003
Translation rate is controlled by coupled trade-offs between site accessibility, selective RNA unfolding and sliding at upstream standby sites Espah Borujeni, Amin; Channarasappa, Anirudh S.; Salis, Howard M. Nucleic Acids Research, Vol. 42, Issue 4 https://doi.org/10.1093/nar/gkt1139	journal	November 2013
Enzymatic Assembly of Overlapping DNA Fragments Gibson, Daniel G. Methods in Enzymology https://doi.org/10.1016/B978-0-12-385120-8.00015-2	book	May 2011
Dcm methylation is detrimental to plasmid transformation in Clostridium thermocellum Guss, Adam M.; Olson, Daniel G.; Caiazza, Nicky C. Biotechnology for Biofuels, Vol. 5, Issue 1 https://doi.org/10.1186/1754-6834-5-30	journal	January 2012
Strain and bioprocess improvement of a thermophilic anaerobe for the production of ethanol from wood Herring, Christopher D.; Kenealy, William R.; Joe Shaw, A. Biotechnology for Biofuels, Vol. 9, Issue 1 https://doi.org/10.1186/s13068-016-0536-8	journal	June 2016
Development and evaluation of methods to infer biosynthesis and substrate consumption in cultures of cellulolytic microorganisms Holwerda, Evert K.; Ellis, Lucas D.; Lynd, Lee R. Biotechnology and Bioengineering, Vol. 110, Issue 9 https://doi.org/10.1002/bit.24915	journal	April 2013
A defined growth medium with very low background carbon for culturing Clostridium thermocellum Holwerda, Evert K.; Hirst, Kyle D.; Lynd, Lee R. Journal of Industrial Microbiology & Biotechnology, Vol. 39, Issue 6 https://doi.org/10.1007/s10295-012-1091-3	journal	February 2012
The exometabolome of Clostridium thermocellum reveals overflow metabolism at high cellulose loading Holwerda, Evert K.; Thorne, Philip G.; Olson, Daniel G. Biotechnology for Biofuels, Vol. 7, Issue 1 https://doi.org/10.1186/s13068-014-0155-1	journal	October 2014
Development of a plasmid-based expression system in Clostridium thermocellum and its use to screen heterologous expression of bifunctional alcohol dehydrogenases (adhEs) Hon, Shuen; Lanahan, Anthony A.; Tian, Liang Metabolic Engineering Communications, Vol. 3 https://doi.org/10.1016/j.meteno.2016.04.001	journal	December 2016
Genetic engineering of Clostridium thermocellum DSM1313 for enhanced ethanol production Kannuchamy, Saranyah; Mukund, Nisha; Saleena, Lilly M. BMC Biotechnology, Vol. 16, Issue S1 https://doi.org/10.1186/s12896-016-0260-2	journal	May 2016
Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2−ΔΔCT Method Livak, Kenneth J.; Schmittgen, Thomas D. Methods, Vol. 25, Issue 4 https://doi.org/10.1006/meth.2001.1262	journal	December 2001
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
The Bifunctional Alcohol and Aldehyde Dehydrogenase Gene, adhE , Is Necessary for Ethanol Production in Clostridium thermocellum and Thermoanaerobacterium saccharolyticum Lo, Jonathan; Zheng, Tianyong; Hon, Shuen Journal of Bacteriology, Vol. 197, Issue 8 https://doi.org/10.1128/JB.02450-14	journal	February 2015
Glycolysis without pyruvate kinase in Clostridium thermocellum Olson, Daniel G.; Hörl, Manuel; Fuhrer, Tobias Metabolic Engineering, Vol. 39 https://doi.org/10.1016/j.ymben.2016.11.011	journal	January 2017
Transformation of Clostridium Thermocellum by Electroporation Olson, Daniel G.; Lynd, Lee R. Cellulases https://doi.org/10.1016/B978-0-12-415931-0.00017-3	book	January 2012
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
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
Ethanol production by engineered thermophiles Olson, Daniel G.; Sparling, Richard; Lynd, Lee R. Current Opinion in Biotechnology, Vol. 33 https://doi.org/10.1016/j.copbio.2015.02.006	journal	June 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
Biological lignocellulose solubilization: comparative evaluation of biocatalysts and enhancement via cotreatment Paye, Julie M. D.; Guseva, Anna; Hammer, Sarah K. Biotechnology for Biofuels, Vol. 9, Issue 1 https://doi.org/10.1186/s13068-015-0412-y	journal	January 2016
Elimination of formate production in Clostridium thermocellum Rydzak, Thomas; Lynd, Lee R.; Guss, Adam M. Journal of Industrial Microbiology & Biotechnology, Vol. 42, Issue 9 https://doi.org/10.1007/s10295-015-1644-3	journal	July 2015
Automated design of synthetic ribosome binding sites to control protein expression Salis, Howard M.; Mirsky, Ethan A.; Voigt, Christopher A. Nature Biotechnology, Vol. 27, Issue 10, p. 946-950 https://doi.org/10.1038/nbt.1568	journal	October 2009
Urease expression in a Thermoanaerobacterium saccharolyticum ethanologen allows high titer ethanol production Joe Shaw, A.; Covalla, Sean F.; Miller, Bethany B. Metabolic Engineering, Vol. 14, Issue 5 https://doi.org/10.1016/j.ymben.2012.06.004	journal	September 2012
Metabolic engineering of a thermophilic bacterium to produce ethanol at high yield Shaw, A. J.; Podkaminer, K. K.; Desai, S. G. Proceedings of the National Academy of Sciences, Vol. 105, Issue 37, p. 13769-13774 https://doi.org/10.1073/pnas.0801266105	journal	September 2008
Ferredoxin:NAD ⁺ Oxidoreductase of Thermoanaerobacterium saccharolyticum and Its Role in Ethanol Formation Tian, Liang; Lo, Jonathan; Shao, Xiongjun Applied and Environmental Microbiology, Vol. 82, Issue 24 https://doi.org/10.1128/AEM.02130-16	journal	September 2016
Simultaneous achievement of high ethanol yield and titer in Clostridium thermocellum Tian, Liang; Papanek, Beth; Olson, Daniel G. Biotechnology for Biofuels, Vol. 9, Issue 1 https://doi.org/10.1186/s13068-016-0528-8	journal	June 2016
Both adhE and a Separate NADPH-Dependent Alcohol Dehydrogenase Gene, adhA , Are Necessary for High Ethanol Production in Thermoanaerobacterium saccharolyticum Zheng, Tianyong; Olson, Daniel G.; Murphy, Sean J. Journal of Bacteriology, Vol. 199, Issue 3 https://doi.org/10.1128/JB.00542-16	journal	November 2016
Cofactor Specificity of the Bifunctional Alcohol and Aldehyde Dehydrogenase (AdhE) in Wild-Type and Mutant Clostridium thermocellum and Thermoanaerobacterium saccharolyticum Zheng, Tianyong; Olson, Daniel G.; Tian, Liang Journal of Bacteriology, Vol. 197, Issue 15 https://doi.org/10.1128/JB.00232-15	journal	May 2015
Hydrogen Formation and Its Regulation in Ruminococcus albus: Involvement of an Electron-Bifurcating [FeFe]-Hydrogenase, of a Non-Electron-Bifurcating [FeFe]-Hydrogenase, and of a Putative Hydrogen-Sensing [FeFe]-Hydrogenase Zheng, Y.; Kahnt, J.; Kwon, I. H. Journal of Bacteriology, Vol. 196, Issue 22 https://doi.org/10.1128/JB.02070-14	journal	August 2014
Physiological roles of pyruvate ferredoxin oxidoreductase and pyruvate formate-lyase in Thermoanaerobacterium saccharolyticum JW/SL-YS485 Zhou, Jilai; Olson, Daniel G.; Lanahan, Anthony A. Biotechnology for Biofuels, Vol. 8, Issue 1 https://doi.org/10.1186/s13068-015-0304-1	journal	September 2015

Cited By (13)

Deletion of the hfsB gene increases ethanol production in Thermoanaerobacterium saccharolyticum and several other thermophilic anaerobic bacteria Eminoğlu, Ayşenur; Murphy, Sean Jean-Loup; Maloney, Marybeth Biotechnology for Biofuels, Vol. 10, Issue 1 https://doi.org/10.1186/s13068-017-0968-9	journal	November 2017
Expressing the Thermoanaerobacterium saccharolyticum pforA in engineered Clostridium thermocellum improves ethanol production Hon, Shuen; Holwerda, Evert K.; Worthen, Robert S. Biotechnology for Biofuels, Vol. 11, Issue 1 https://doi.org/10.1186/s13068-018-1245-2	journal	September 2018
Metabolome analysis reveals a role for glyceraldehyde 3-phosphate dehydrogenase in the inhibition of C. thermocellum by ethanol Tian, Liang; Perot, Skyler J.; Stevenson, David Biotechnology for Biofuels, Vol. 10, Issue 1 https://doi.org/10.1186/s13068-017-0961-3	journal	November 2017
Metabolic and evolutionary responses of Clostridium thermocellum to genetic interventions aimed at improving ethanol production Holwerda, Evert K.; Olson, Daniel G.; Ruppertsberger, Natalie M. Biotechnology for Biofuels, Vol. 13, Issue 1 https://doi.org/10.1186/s13068-020-01680-5	journal	March 2020
Enhanced ethanol formation by Clostridium thermocellum via pyruvate decarboxylase Tian, Liang; Perot, Skyler J.; Hon, Shuen Microbial Cell Factories, Vol. 16, Issue 1 https://doi.org/10.1186/s12934-017-0783-9	journal	October 2017
Metabolic engineering of Clostridium thermocellum for n-butanol production from cellulose Tian, Liang; Conway, Peter M.; Cervenka, Nicholas D. Biotechnology for Biofuels, Vol. 12, Issue 1 https://doi.org/10.1186/s13068-019-1524-6	journal	July 2019
Building a genome engineering toolbox in nonmodel prokaryotic microbes Freed, Emily; Fenster, Jacob; Smolinski, Sharon L. Biotechnology and Bioengineering, Vol. 115, Issue 9 https://doi.org/10.1002/bit.26727	journal	May 2018
A mutation in the AdhE alcohol dehydrogenase of Clostridium thermocellum increases tolerance to several primary alcohols, including isobutanol, n-butanol and ethanol Tian, Liang; Cervenka, Nicholas D.; Low, Aidan M. Scientific Reports, Vol. 9, Issue 1 https://doi.org/10.1038/s41598-018-37979-5	journal	February 2019
Clostridium thermocellum LL1210 pH homeostasis mechanisms informed by transcriptomics and metabolomics Whitham, Jason M.; Moon, Ji-Won; Rodriguez, Miguel Biotechnology for Biofuels, Vol. 11, Issue 1 https://doi.org/10.1186/s13068-018-1095-y	journal	April 2018
Expression of adhA from different organisms in Clostridium thermocellum Zheng, Tianyong; Cui, Jingxuan; Bae, Hye Ri Biotechnology for Biofuels, Vol. 10, Issue 1 https://doi.org/10.1186/s13068-017-0940-8	journal	November 2017
Engineering Clostridium for improved solvent production: recent progress and perspective Cheng, Chi; Bao, Teng; Yang, Shang-Tian Applied Microbiology and Biotechnology, Vol. 103, Issue 14 https://doi.org/10.1007/s00253-019-09916-7	journal	May 2019
Recent advances of biofuels and biochemicals production from sustainable resources using co-cultivation systems Jiang, Yujia; Wu, Ruofan; Zhou, Jie Biotechnology for Biofuels, Vol. 12, Issue 1 https://doi.org/10.1186/s13068-019-1495-7	journal	June 2019
The redox-sensing protein Rex modulates ethanol production in Thermoanaerobacterium saccharolyticum Zheng, Tianyong; Lanahan, Anthony A.; Lynd, Lee R. PLOS ONE, Vol. 13, Issue 4 https://doi.org/10.1371/journal.pone.0195143	journal	April 2018

Similar Records

Expressing the Thermoanaerobacterium saccharolyticum pforA in engineered Clostridium thermocellum improves ethanol production

Journal Article · Thu Sep 06 00:00:00 EDT 2018 · Biotechnology for Biofuels · OSTI ID:1394382

Hon, Shuen; Holwerda, Evert K.; Worthen, Robert S.; +6 more

Cofactor Specificity of the Bifunctional Alcohol and Aldehyde Dehydrogenase (AdhE) in Wild-Type and Mutant Clostridium thermocellum and Thermoanaerobacterium saccharolyticum

Journal Article · Tue May 26 00:00:00 EDT 2015 · Journal of Bacteriology · OSTI ID:1394382

Zheng, Tianyong; Olson, Daniel G.; Tian, Liang; +8 more

In Vivo Thermodynamic Analysis of Glycolysis in Clostridium thermocellum and Thermoanaerobacterium saccharolyticum Using ¹³ C and ² H Tracers

Journal Article · Tue Mar 17 00:00:00 EDT 2020 · mSystems · OSTI ID:1394382

Jacobson, Tyler B.; Korosh, Travis K.; Stevenson, David M.; +6 more

Related Subjects

09 BIOMASS FUELS
59 BASIC BIOLOGICAL SCIENCES
60 APPLIED LIFE SCIENCES

Title: The ethanol pathway from Thermoanaerobacterium saccharolyticum improves ethanol production in Clostridium thermocellum

Citation Formats

References (34)

Cited By (13)

Similar Records

Related Subjects