Improved growth rate in Clostridium thermocellum hydrogenase mutant via perturbed sulfur metabolism
Abstract
Background: Metabolic engineering is a commonly used approach to develop organisms for an industrial function, but engineering aimed at improving one phenotype can negatively impact other phenotypes. This lack of robustness can prove problematic. Cellulolytic bacterium Clostridium thermocellum is able to rapidly ferment cellulose to ethanol and other products. Recently, genes involved in H2 production, including the hydrogenase maturase hydG, were deleted from the chromosome of C. thermocellum. While ethanol yield increased, the growth rate decreased substantially compared to wild type. Results: Addition of 5 mM acetate to the growth medium improved the growth rate in C. thermocellum ΔhydG, whereas wild type remained unaffected. Transcriptomic analysis of the wild type showed essentially no response to the addition of acetate. However, in C. thermocellum ΔhydG, 204 and 56 genes were significantly differentially regulated relative to wild type in the absence and presence of acetate, respectively. Genes Clo1313_0108-0125, which are predicted to encode a sulfate transport system and sulfate assimilatory pathway, were drastically up-regulated in C. thermocellum ΔhydG in presence of added acetate. A similar pattern was seen with proteomics. Further physiological characterization demonstrated an increase in sulfide synthesis and elimination of cysteine consumption in C. thermocellum ΔhydG. In conclusion, sulfur metabolismmore »
- 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:
- 1618660
- Alternate Identifier(s):
- OSTI ID: 1393809
- Grant/Contract Number:
- BioEnergy Science Center; AC05-00OR22725
- Resource Type:
- Published Article
- Journal Name:
- Biotechnology for Biofuels
- Additional Journal Information:
- Journal Name: Biotechnology for Biofuels Journal Volume: 10 Journal Issue: 1; Journal ID: ISSN 1754-6834
- Publisher:
- Springer Science + Business Media
- Country of Publication:
- Netherlands
- Language:
- English
- Subject:
- 09 BIOMASS FUELS; Cellulosic ethanol; Clostridium thermocellum; Redox balance; Metabolic engineering; Sulfate reduction
Citation Formats
Biswas, Ranjita, Wilson, Charlotte M., Giannone, Richard J., Klingeman, Dawn M., Rydzak, Thomas, Shah, Manesh B., Hettich, Robert L., Brown, Steven D., and Guss, Adam M. Improved growth rate in Clostridium thermocellum hydrogenase mutant via perturbed sulfur metabolism. Netherlands: N. p., 2017.
Web. doi:10.1186/s13068-016-0684-x.
Biswas, Ranjita, Wilson, Charlotte M., Giannone, Richard J., Klingeman, Dawn M., Rydzak, Thomas, Shah, Manesh B., Hettich, Robert L., Brown, Steven D., & Guss, Adam M. Improved growth rate in Clostridium thermocellum hydrogenase mutant via perturbed sulfur metabolism. Netherlands. https://doi.org/10.1186/s13068-016-0684-x
Biswas, Ranjita, Wilson, Charlotte M., Giannone, Richard J., Klingeman, Dawn M., Rydzak, Thomas, Shah, Manesh B., Hettich, Robert L., Brown, Steven D., and Guss, Adam M. Tue .
"Improved growth rate in Clostridium thermocellum hydrogenase mutant via perturbed sulfur metabolism". Netherlands. https://doi.org/10.1186/s13068-016-0684-x.
@article{osti_1618660,
title = {Improved growth rate in Clostridium thermocellum hydrogenase mutant via perturbed sulfur metabolism},
author = {Biswas, Ranjita and Wilson, Charlotte M. and Giannone, Richard J. and Klingeman, Dawn M. and Rydzak, Thomas and Shah, Manesh B. and Hettich, Robert L. and Brown, Steven D. and Guss, Adam M.},
abstractNote = {Background: Metabolic engineering is a commonly used approach to develop organisms for an industrial function, but engineering aimed at improving one phenotype can negatively impact other phenotypes. This lack of robustness can prove problematic. Cellulolytic bacterium Clostridium thermocellum is able to rapidly ferment cellulose to ethanol and other products. Recently, genes involved in H2 production, including the hydrogenase maturase hydG, were deleted from the chromosome of C. thermocellum. While ethanol yield increased, the growth rate decreased substantially compared to wild type. Results: Addition of 5 mM acetate to the growth medium improved the growth rate in C. thermocellum ΔhydG, whereas wild type remained unaffected. Transcriptomic analysis of the wild type showed essentially no response to the addition of acetate. However, in C. thermocellum ΔhydG, 204 and 56 genes were significantly differentially regulated relative to wild type in the absence and presence of acetate, respectively. Genes Clo1313_0108-0125, which are predicted to encode a sulfate transport system and sulfate assimilatory pathway, were drastically up-regulated in C. thermocellum ΔhydG in presence of added acetate. A similar pattern was seen with proteomics. Further physiological characterization demonstrated an increase in sulfide synthesis and elimination of cysteine consumption in C. thermocellum ΔhydG. In conclusion, sulfur metabolism is perturbed in C. thermocellum ΔhydG, possibly to increase flux through sulfate reduction to act as an electron sink to balance redox reactions.},
doi = {10.1186/s13068-016-0684-x},
journal = {Biotechnology for Biofuels},
number = 1,
volume = 10,
place = {Netherlands},
year = {Tue Jan 03 00:00:00 EST 2017},
month = {Tue Jan 03 00:00:00 EST 2017}
}
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https://doi.org/10.1186/s13068-016-0684-x
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Cited by: 7 works
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Figures / Tables:
Fig. 1: Growth profle of C. thermocellum strains on minimal medium. a Wild type, b ΔhydG, and c ΔhydG Δech. Symbols: red square, with added acetate; black triangle, without added acetate. Data for “without added acetate” are from [11]
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