The redox-sensing protein Rex modulates ethanol production in Thermoanaerobacterium saccharolyticum
Abstract
Thermoanaerobacterium saccharolyticum is a thermophilic anaerobe that has been engineered to produce high amounts of ethanol, reaching ~90% theoretical yield at a titer of 70 g/L. Here we report the physiological changes that occur upon deleting the redox-sensing transcriptional regulator Rex in wild type T. saccharolyticum: a single deletion of rex resulted in a two-fold increase in ethanol yield (from 40% to 91% theoretical yield), but the resulting strains grew only about a third as fast as the wild type strain. Deletion of the rex gene also had the effect of increasing expression of alcohol dehydrogenase genes, adhE and adhA. After several serial transfers, the ethanol yield decreased from an average of 91% to 55%, and the growth rates had increased. We performed whole-genome resequencing to identify secondary mutations in the Δrex strains adapted for faster growth. In several cases, secondary mutations had appeared in the adhE gene. Furthermore, in these strains the NADH-linked alcohol dehydrogenase activity was greatly reduced. Complementation studies were done to reintroduce rex into the Δ strains: reintroducing rex decreased ethanol yield to below wild type levels in the Δrex strain without adhE mutations, but did not change the ethanol yield in the Δrex strain wheremore »
- Authors:
- Publication Date:
- Research Org.:
- Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Biological and Environmental Research (BER)
- OSTI Identifier:
- 1432066
- Alternate Identifier(s):
- OSTI ID: 1627859
- Grant/Contract Number:
- AC05-00OR22725
- Resource Type:
- Published Article
- Journal Name:
- PLoS ONE
- Additional Journal Information:
- Journal Name: PLoS ONE Journal Volume: 13 Journal Issue: 4; Journal ID: ISSN 1932-6203
- Publisher:
- Public Library of Science
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 09 BIOMASS FUELS; Science & Technology - Other Topics
Citation Formats
Zheng, Tianyong, Lanahan, Anthony A., Lynd, Lee R., Olson, Daniel G., and Blanchard, ed., Jeffrey L. The redox-sensing protein Rex modulates ethanol production in Thermoanaerobacterium saccharolyticum. United States: N. p., 2018.
Web. doi:10.1371/journal.pone.0195143.
Zheng, Tianyong, Lanahan, Anthony A., Lynd, Lee R., Olson, Daniel G., & Blanchard, ed., Jeffrey L. The redox-sensing protein Rex modulates ethanol production in Thermoanaerobacterium saccharolyticum. United States. https://doi.org/10.1371/journal.pone.0195143
Zheng, Tianyong, Lanahan, Anthony A., Lynd, Lee R., Olson, Daniel G., and Blanchard, ed., Jeffrey L. Thu .
"The redox-sensing protein Rex modulates ethanol production in Thermoanaerobacterium saccharolyticum". United States. https://doi.org/10.1371/journal.pone.0195143.
@article{osti_1432066,
title = {The redox-sensing protein Rex modulates ethanol production in Thermoanaerobacterium saccharolyticum},
author = {Zheng, Tianyong and Lanahan, Anthony A. and Lynd, Lee R. and Olson, Daniel G. and Blanchard, ed., Jeffrey L.},
abstractNote = {Thermoanaerobacterium saccharolyticum is a thermophilic anaerobe that has been engineered to produce high amounts of ethanol, reaching ~90% theoretical yield at a titer of 70 g/L. Here we report the physiological changes that occur upon deleting the redox-sensing transcriptional regulator Rex in wild type T. saccharolyticum: a single deletion of rex resulted in a two-fold increase in ethanol yield (from 40% to 91% theoretical yield), but the resulting strains grew only about a third as fast as the wild type strain. Deletion of the rex gene also had the effect of increasing expression of alcohol dehydrogenase genes, adhE and adhA. After several serial transfers, the ethanol yield decreased from an average of 91% to 55%, and the growth rates had increased. We performed whole-genome resequencing to identify secondary mutations in the Δrex strains adapted for faster growth. In several cases, secondary mutations had appeared in the adhE gene. Furthermore, in these strains the NADH-linked alcohol dehydrogenase activity was greatly reduced. Complementation studies were done to reintroduce rex into the Δ strains: reintroducing rex decreased ethanol yield to below wild type levels in the Δrex strain without adhE mutations, but did not change the ethanol yield in the Δrex strain where an adhE mutation occurred.},
doi = {10.1371/journal.pone.0195143},
journal = {PLoS ONE},
number = 4,
volume = 13,
place = {United States},
year = {Thu Apr 05 00:00:00 EDT 2018},
month = {Thu Apr 05 00:00:00 EDT 2018}
}
https://doi.org/10.1371/journal.pone.0195143
Web of Science
Figures / Tables:
Works referenced in this record:
The ethanol stress response and ethanol tolerance of Saccharomyces cerevisiae
journal, January 2010
- Stanley, D.; Bandara, A.; Fraser, S.
- Journal of Applied Microbiology
Profile of Secreted Hydrolases, Associated Proteins, and SlpA in Thermoanaerobacterium saccharolyticum during the Degradation of Hemicellulose
journal, June 2014
- Currie, D. H.; Guss, A. M.; Herring, C. D.
- Applied and Environmental Microbiology, Vol. 80, Issue 16
Ferredoxin:NAD + Oxidoreductase of Thermoanaerobacterium saccharolyticum and Its Role in Ethanol Formation
journal, September 2016
- Tian, Liang; Lo, Jonathan; Shao, Xiongjun
- Applied and Environmental Microbiology, Vol. 82, Issue 24
Redox sensing by a Rex-family repressor is involved in the regulation of anaerobic gene expression in Staphylococcus aureus: Redox sensitive gene regulation in S. aureus
journal, March 2010
- Pagels, Martin; Fuchs, Stephan; Pané-Farré, Jan
- Molecular Microbiology, Vol. 76, Issue 5
Ethanol Tolerance in Bacteria
journal, January 1989
- Ingram, Lonnie O.
- Critical Reviews in Biotechnology, Vol. 9, Issue 4
Analysis of composition and structure of Clostridium thermocellum membranes from wild-type and ethanol-adapted strains
journal, February 2009
- Timmons, Michael D.; Knutson, Barbara L.; Nokes, Sue E.
- Applied Microbiology and Biotechnology, Vol. 82, Issue 5
Genome-scale resources for Thermoanaerobacterium saccharolyticum
journal, June 2015
- Currie, Devin H.; Raman, Babu; Gowen, Christopher M.
- BMC Systems Biology, Vol. 9, Issue 1
Natural Competence in Thermoanaerobacter and Thermoanaerobacterium Species
journal, May 2010
- Shaw, A.; Hogsett, D.; Lynd, L.
- Applied and Environmental Microbiology, Vol. 76, Issue 14, p. 4713-4719
The redox-sensing protein Rex, a transcriptional regulator of solventogenesis in Clostridium acetobutylicum
journal, May 2012
- Wietzke, Mandy; Bahl, Hubert
- Applied Microbiology and Biotechnology, Vol. 96, Issue 3
Energy conservation via electron bifurcating ferredoxin reduction and proton/Na+ translocating ferredoxin oxidation
journal, February 2013
- Buckel, Wolfgang; Thauer, Rudolf K.
- Biochimica et Biophysica Acta (BBA) - Bioenergetics, Vol. 1827, Issue 2
The Redox-Sensing Regulator Rex Modulates Central Carbon Metabolism, Stress Tolerance Response and Biofilm Formation by Streptococcus mutans
journal, September 2012
- Bitoun, Jacob P.; Liao, Sumei; Yao, Xin
- PLoS ONE, Vol. 7, Issue 9
Genetic Dissection of Ethanol Tolerance in the Budding Yeast Saccharomyces cerevisiae
journal, December 2006
- Hu, X. H.; Wang, M. H.; Tan, T.
- Genetics, Vol. 175, Issue 3
Effects of ethanol and other alkanols on the general amino acid permease of Saccharomyces cerevisiae : COMMUNICATIONS TO THE EDITOR
journal, April 1984
- Leão, Cecilia; van Uden, N.
- Biotechnology and Bioengineering, Vol. 26, Issue 4
Physiological roles of pyruvate ferredoxin oxidoreductase and pyruvate formate-lyase in Thermoanaerobacterium saccharolyticum JW/SL-YS485
journal, September 2015
- Zhou, Jilai; Olson, Daniel G.; Lanahan, Anthony A.
- Biotechnology for Biofuels, Vol. 8, Issue 1
Tolerance and stress response to ethanol in the yeast Saccharomyces cerevisiae
journal, September 2009
- Ding, Junmei; Huang, Xiaowei; Zhang, Lemin
- Applied Microbiology and Biotechnology, Vol. 85, Issue 2
Transcriptional Regulation of Central Carbon and Energy Metabolism in Bacteria by Redox-Responsive Repressor Rex
journal, December 2011
- Ravcheev, D. A.; Li, X.; Latif, H.
- Journal of Bacteriology, Vol. 194, Issue 5
Transcriptional repressor Rex is involved in regulation of oxidative stress response and biofilm formation by Streptococcus mutans: Transcriptional repressor Rex in Streptococcus mutans
journal, May 2011
- Bitoun, Jacob P.; Nguyen, Anne H.; Fan, Yuwei
- FEMS Microbiology Letters, Vol. 320, Issue 2
Transformation of Thermoanaerobacterium sp. strain JW/SL-YS485 with plasmid pIKM1 conferring kanamycin resistance
journal, March 1997
- Mai, Volker; Lorenz, W. Walter; Wiegel, Juergen
- FEMS Microbiology Letters, Vol. 148, Issue 2
End-product pathways in the xylose fermenting bacterium, Thermoanaerobacterium saccharolyticum
journal, May 2008
- Joe Shaw, A.; Jenney, Francis E.; Adams, Michael W. W.
- Enzyme and Microbial Technology, Vol. 42, Issue 6
Pattern locator: a new tool for finding local sequence patterns in genomic DNA sequences
journal, November 2006
- Mrazek, J.; Xie, S.
- Bioinformatics, Vol. 22, Issue 24
Both adhE and a Separate NADPH-Dependent Alcohol Dehydrogenase Gene, adhA , Are Necessary for High Ethanol Production in Thermoanaerobacterium saccharolyticum
journal, November 2016
- Zheng, Tianyong; Olson, Daniel G.; Murphy, Sean J.
- Journal of Bacteriology, Vol. 199, Issue 3
Structural Basis for NADH/NAD+ Redox Sensing by a Rex Family Repressor
journal, May 2010
- McLaughlin, Krystle J.; Strain-Damerell, Claire M.; Xie, Kefang
- Molecular Cell, Vol. 38, Issue 4
Metabolic engineering of a thermophilic bacterium to produce ethanol at high yield
journal, September 2008
- Shaw, A. J.; Podkaminer, K. K.; Desai, S. G.
- Proceedings of the National Academy of Sciences, Vol. 105, Issue 37, p. 13769-13774
High Ethanol Titers from Cellulose by Using Metabolically Engineered Thermophilic, Anaerobic Microbes
journal, September 2011
- Argyros, D. Aaron; Tripathi, Shital A.; Barrett, Trisha F.
- Applied and Environmental Microbiology, Vol. 77, Issue 23, p. 8288-8294
The mechanism for regulating ethanol fermentation by redox levels in Thermoanaerobacter ethanolicus
journal, March 2011
- Pei, Jianjun; Zhou, Qing; Jing, Qingqing
- Metabolic Engineering, Vol. 13, Issue 2
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