Expression of a heat-stable NADPH-dependent alcohol dehydrogenase from Thermoanaerobacter pseudethanolicus 39E in Clostridium thermocellum 1313 results in increased hydroxymethylfurfural resistance
Abstract
Background: 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. Results: Heterologous expression of the Thermoanaerobacter pseudethanolicus 39E bdhA gene, that encodes an aldehyde dehydrogenase, in Clostridium thermocellum significantly increased resistance to furan derivatives at concentrations found in acid-pretreated biomass. The mechanism of detoxification of HMF 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. Conclusions: Toxic compounds present in lignocellulose hydrolysates that inhibit cell growth and product formation are obstacles to the commercialization of fuels and chemicals from biomass. Expression of genes that reduce the effectmore »
- Authors:
-
- 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)
- Publication Date:
- Research Org.:
- National Renewable Energy Laboratory (NREL), Golden, CO (United States); Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center (BESC)
- Sponsoring Org.:
- USDOE Office of Science (SC), Biological and Environmental Research (BER)
- OSTI Identifier:
- 1350020
- Alternate Identifier(s):
- OSTI ID: 1361373
- Report Number(s):
- NREL/JA-2700-68259
Journal ID: ISSN 1754-6834
- Grant/Contract Number:
- AC36-08GO28308; AC05-00OR22725
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Biotechnology for Biofuels
- Additional Journal Information:
- Journal Volume: 10; Journal Issue: 1; Journal ID: ISSN 1754-6834
- Publisher:
- BioMed Central
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 09 BIOMASS FUELS; consolidated bioprocessing; Clostridium thermocellum; butanol dehydrogenase; furfural; 5-hydroxymethyl-2-furfural
Citation Formats
Kim, Sun -Ki, Groom, Joseph, Chung, Daehwan, Elkins, James, and Westpheling, Janet. Expression of a heat-stable NADPH-dependent alcohol dehydrogenase from Thermoanaerobacter pseudethanolicus 39E in Clostridium thermocellum 1313 results in increased hydroxymethylfurfural resistance. United States: N. p., 2017.
Web. doi:10.1186/s13068-017-0750-z.
Kim, Sun -Ki, Groom, Joseph, Chung, Daehwan, Elkins, James, & Westpheling, Janet. Expression of a heat-stable NADPH-dependent alcohol dehydrogenase from Thermoanaerobacter pseudethanolicus 39E in Clostridium thermocellum 1313 results in increased hydroxymethylfurfural resistance. United States. https://doi.org/10.1186/s13068-017-0750-z
Kim, Sun -Ki, Groom, Joseph, Chung, Daehwan, Elkins, James, and Westpheling, Janet. Wed .
"Expression of a heat-stable NADPH-dependent alcohol dehydrogenase from Thermoanaerobacter pseudethanolicus 39E in Clostridium thermocellum 1313 results in increased hydroxymethylfurfural resistance". United States. https://doi.org/10.1186/s13068-017-0750-z. https://www.osti.gov/servlets/purl/1350020.
@article{osti_1350020,
title = {Expression of a heat-stable NADPH-dependent alcohol dehydrogenase from Thermoanaerobacter pseudethanolicus 39E in Clostridium thermocellum 1313 results in increased hydroxymethylfurfural resistance},
author = {Kim, Sun -Ki and Groom, Joseph and Chung, Daehwan and Elkins, James and Westpheling, Janet},
abstractNote = {Background: 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. Results: Heterologous expression of the Thermoanaerobacter pseudethanolicus 39E bdhA gene, that encodes an aldehyde dehydrogenase, in Clostridium thermocellum significantly increased resistance to furan derivatives at concentrations found in acid-pretreated biomass. The mechanism of detoxification of HMF 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. Conclusions: Toxic compounds present in lignocellulose hydrolysates that inhibit cell growth and product formation are obstacles to the commercialization of fuels and chemicals from biomass. 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.},
doi = {10.1186/s13068-017-0750-z},
journal = {Biotechnology for Biofuels},
number = 1,
volume = 10,
place = {United States},
year = {Wed Mar 15 00:00:00 EDT 2017},
month = {Wed Mar 15 00:00:00 EDT 2017}
}
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Cited by: 13 works
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Figures / Tables:
Fig. 1: Maps of shuttle vectors for BdhA expression in $C. thermocellum$. The bdhA gene from $T. pseudethanolicus$ 39E (Teth39_1597) was expressed under the control of the S‑layer (a), Clo1313_1809 (b), and enolase (c) promoters. Shuttle vectors contain a C‑terminal 6X His‑tag version of bdhA, a Rho‑independent terminator, the pyrFmore »
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Increased tolerance and conversion of inhibitors in lignocellulosic hydrolysates by Saccharomyces cerevisiae
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NADH- vs NADPH-coupled reduction of 5-hydroxymethyl furfural (HMF) and its implications on product distribution in Saccharomyces cerevisiae
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Multiple gene-mediated NAD(P)H-dependent aldehyde reduction is a mechanism of in situ detoxification of furfural and 5-hydroxymethylfurfural by Saccharomyces cerevisiae
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Effect of dilute acid pretreatment of rice straw on structural properties and enzymatic hydrolysis
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Expression of aldehyde dehydrogenase 6 reduces inhibitory effect of furan derivatives on cell growth and ethanol production in Saccharomyces cerevisiae
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Recent progress in consolidated bioprocessing
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Development of a high-throughput method to evaluate the impact of inhibitory compounds from lignocellulosic hydrolysates on the growth of Zymomonas mobilis
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Identifying promoters for gene expression in Clostridium thermocellum
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Promiscuous plasmid replication in thermophiles: Use of a novel hyperthermophilic replicon for genetic manipulation of Clostridium thermocellum at its optimum growth temperature
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Redirecting carbon flux through exogenous pyruvate kinase to achieve high ethanol yields in Clostridium thermocellum
journal, January 2013
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Enhanced tolerance of Saccharomyces cerevisiae to multiple lignocellulose-derived inhibitors through modulation of spermidine contents
journal, May 2015
- Kim, Sun-Ki; Jin, Yong-Su; Choi, In-Geol
- Metabolic Engineering, Vol. 29
Engineering electron metabolism to increase ethanol production in Clostridium thermocellum
journal, January 2017
- Lo, Jonathan; Olson, Daniel G.; Murphy, Sean Jean-Loup
- Metabolic Engineering, Vol. 39
Mutant alcohol dehydrogenase leads to improved ethanol tolerance in Clostridium thermocellum
journal, August 2011
- Brown, S. D.; Guss, A. M.; Karpinets, T. V.
- Proceedings of the National Academy of Sciences, Vol. 108, Issue 33
Engineering furfural tolerance in Escherichia coli improves the fermentation of lignocellulosic sugars into renewable chemicals
journal, February 2013
- Wang, X.; Yomano, L. P.; Lee, J. Y.
- Proceedings of the National Academy of Sciences, Vol. 110, Issue 10, p. 4021-4026
Direct conversion of plant biomass to ethanol by engineered Caldicellulosiruptor bescii
journal, June 2014
- Chung, D.; Cha, M.; Guss, A. M.
- Proceedings of the National Academy of Sciences, Vol. 111, Issue 24, p. 8931-8936
Insights into plant biomass conversion from the genome of the anaerobic thermophilic bacterium Caldicellulosiruptor bescii DSM 6725
journal, January 2011
- Dam, Phuongan; Kataeva, Irina; Yang, Sung-Jae
- Nucleic Acids Research, Vol. 39, Issue 8
A rapid chemical method for lysing Arabidopsis cells for protein analysis
journal, January 2011
- Tsugama, Daisuke; Liu, Shenkui; Takano, Tetsuo
- Plant Methods, Vol. 7, Issue 1
The influence of HMF and furfural on redox-balance and energy-state of xylose-utilizing Saccharomyces cerevisiae
journal, January 2013
- Ask, Magnus; Bettiga, Maurizio; Mapelli, Valeria
- Biotechnology for Biofuels, Vol. 6, Issue 1
Elimination of hydrogenase active site assembly blocks H2 production and increases ethanol yield in Clostridium thermocellum
journal, January 2015
- Biswas, Ranjita; Zheng, Tianyong; Olson, Daniel G.
- Biotechnology for Biofuels, Vol. 8, Issue 1
Expression of a heat-stable NADPH-dependent alcohol dehydrogenase in Caldicellulosiruptor bescii results in furan aldehyde detoxification
journal, July 2015
- Chung, Daehwan; Verbeke, Tobin J.; Cross, Karissa L.
- Biotechnology for Biofuels, Vol. 8, Issue 1
The emergence of Clostridium thermocellum as a high utility candidate for consolidated bioprocessing applications
journal, August 2014
- Akinosho, Hannah; Yee, Kelsey; Close, Dan
- Frontiers in Chemistry, Vol. 2
Works referencing / citing this record:
Firmicutes-enriched IS1447 represents a group of IS3-family insertion sequences exhibiting unique + 1 transcriptional slippage
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Figures / Tables found in this record:
Figures/Tables have been extracted from DOE-funded journal article accepted manuscripts.