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Title: Metabolic and evolutionary responses of Clostridium thermocellum to genetic interventions aimed at improving ethanol production

Abstract

Engineering efforts targeted at increasing ethanol by modifying the central fermentative metabolism of Clostridium thermocellum have been variably successful. Here, we aim to understand this variation by a multifaceted approach including genomic and transcriptomic analysis combined with chemostat cultivation and high solids cellulose fermentation. Three strain lineages comprising 16 strains total were examined. Two strain lineages in which genes involved in pathways leading to organic acids and/or sporulation had been knocked out resulted in four end-strains after adaptive laboratory evolution (ALE). A third strain lineage recapitulated mutations involving adhE that occurred spontaneously in some of the engineered strains.

Authors:
ORCiD logo [1];  [1];  [2];  [3];  [2];  [1];  [1];  [3];  [1]
  1. Dartmouth College, Hanover, NH (United States). Thayer School of Engineering; Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Center for Bioenergy Innovation
  2. Dartmouth College, Hanover, NH (United States). Thayer School of Engineering
  3. Univ. of Wisconsin, Madison, WI (United States). Dept. of Bacteriology; Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Center for Bioenergy Innovation
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER)
OSTI Identifier:
1627007
Grant/Contract Number:  
AC02-05CH11231
Resource Type:
Accepted Manuscript
Journal Name:
Biotechnology for Biofuels
Additional Journal Information:
Journal Volume: 13; Journal Issue: 1; Journal ID: ISSN 1754-6834
Publisher:
BioMed Central
Country of Publication:
United States
Language:
English
Subject:
Biotechnology & Applied Microbiology; Energy & Fuels

Citation Formats

Holwerda, Evert K., Olson, Daniel G., Ruppertsberger, Natalie M., Stevenson, David M., Murphy, Sean J. L., Maloney, Marybeth I., Lanahan, Anthony A., Amador-Noguez, Daniel, and Lynd, Lee R. Metabolic and evolutionary responses of Clostridium thermocellum to genetic interventions aimed at improving ethanol production. United States: N. p., 2020. Web. https://doi.org/10.1186/s13068-020-01680-5.
Holwerda, Evert K., Olson, Daniel G., Ruppertsberger, Natalie M., Stevenson, David M., Murphy, Sean J. L., Maloney, Marybeth I., Lanahan, Anthony A., Amador-Noguez, Daniel, & Lynd, Lee R. Metabolic and evolutionary responses of Clostridium thermocellum to genetic interventions aimed at improving ethanol production. United States. https://doi.org/10.1186/s13068-020-01680-5
Holwerda, Evert K., Olson, Daniel G., Ruppertsberger, Natalie M., Stevenson, David M., Murphy, Sean J. L., Maloney, Marybeth I., Lanahan, Anthony A., Amador-Noguez, Daniel, and Lynd, Lee R. Tue . "Metabolic and evolutionary responses of Clostridium thermocellum to genetic interventions aimed at improving ethanol production". United States. https://doi.org/10.1186/s13068-020-01680-5. https://www.osti.gov/servlets/purl/1627007.
@article{osti_1627007,
title = {Metabolic and evolutionary responses of Clostridium thermocellum to genetic interventions aimed at improving ethanol production},
author = {Holwerda, Evert K. and Olson, Daniel G. and Ruppertsberger, Natalie M. and Stevenson, David M. and Murphy, Sean J. L. and Maloney, Marybeth I. and Lanahan, Anthony A. and Amador-Noguez, Daniel and Lynd, Lee R.},
abstractNote = {Engineering efforts targeted at increasing ethanol by modifying the central fermentative metabolism of Clostridium thermocellum have been variably successful. Here, we aim to understand this variation by a multifaceted approach including genomic and transcriptomic analysis combined with chemostat cultivation and high solids cellulose fermentation. Three strain lineages comprising 16 strains total were examined. Two strain lineages in which genes involved in pathways leading to organic acids and/or sporulation had been knocked out resulted in four end-strains after adaptive laboratory evolution (ALE). A third strain lineage recapitulated mutations involving adhE that occurred spontaneously in some of the engineered strains.},
doi = {10.1186/s13068-020-01680-5},
journal = {Biotechnology for Biofuels},
number = 1,
volume = 13,
place = {United States},
year = {2020},
month = {3}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record

Figures / Tables:

Fig. 1 Fig. 1: Overview of the strain lineages and data associated with each strain (a) and overview of the research approach and data analysis in this paper (b). A total of 16 strains were analyzed, covering three different lineages. For each stage of the analysis different data sets were collected. Fourmore » strains (LL1011, LL1043, LL374 and LL375) have been subjected to Adaptive Laboratory Evolution, as indicated in the figure. For description of the strain, the adaptation, data generation and analysis see “Methods” section« less

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