Elimination of hydrogenase active site assembly blocks H2 production and increases ethanol yield in Clostridium thermocellum

Biswas, Ranjita; Zheng, Tianyong; Olson, Daniel G.; Lynd, Lee R.; Guss, Adam M.

doi:10.1186/s13068-015-0204-4

Title: Elimination of hydrogenase active site assembly blocks H2 production and increases ethanol yield in Clostridium thermocellum

Journal Article · Thu Feb 12 00:00:00 EST 2015 · Biotechnology for Biofuels

DOI:https://doi.org/10.1186/s13068-015-0204-4· OSTI ID:1185431

Biswas, Ranjita ^[1]; Zheng, Tianyong ^[2]; Olson, Daniel G. ^[2]; Lynd, Lee R. ^[2]; Guss, Adam M. ^[1]

Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Biosciences Div. and BioEnergy Science Center
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center; Dartmouth College, Hanover, NH (United States). Thayer School of Engineering

Background: The native ability of Clostridium thermocellum to rapidly consume cellulose and produce ethanol makes it a leading candidate for a consolidated bioprocessing (CBP) biofuel production strategy. C. thermocellum also synthesizes lactate, formate, acetate, H-2, and amino acids that compete with ethanol production for carbon and electrons. Elimination of H-2 production could redirect carbon flux towards ethanol production by making more electrons available for acetyl coenzyme A reduction to ethanol. Results: H-2 production in C. thermocellum is encoded by four hydrogenases. Rather than delete each individually, we targeted hydrogenase maturase gene hydG, involved in converting the three [FeFe] hydrogenase apoenzymes into holoenzymes. Further deletion of the [NiFe] hydrogenase (ech) resulted in a mutant that functionally lacks all four hydrogenases. H-2 production in ΔhydGΔech was undetectable, and the ethanol yield nearly doubled to 64% of the maximum theoretical yield. Genomic analysis of ΔhydG revealed a mutation in adhE, resulting in a strain with both NADH- and NADPH-dependent alcohol dehydrogenase activities. While this same adhE mutation was found in ethanol-tolerant C. thermocellum strain E50C, ΔhydG and ΔhydGΔech are not more ethanol tolerant than the wild type, illustrating the complicated interactions between redox balancing and ethanol tolerance in C. thermocellum. Conclusions: Finally, the dramatic increase in ethanol production suggests that targeting protein post-translational modification is a promising new approach for simultaneous inactivation of multiple enzymes.

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Research Organization:: Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center (BESC)

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

Grant/Contract Number:: AC05-00OR22725

OSTI ID:: 1185431

Alternate ID(s):: OSTI ID: 1327623

Journal Information:: Biotechnology for Biofuels, Vol. 8, Issue 1; ISSN 1754-6834

Publisher:: BioMed CentralCopyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 82 works

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