The exometabolome of Clostridium thermocellum reveals overflow metabolism at high cellulose loading
Abstract
Background: Clostridium thermocellum is a model thermophilic organism for the production of biofuels from lignocellulosic substrates. The majority of publications studying the physiology of this organism use substrate concentrations of ≤10 g/L. However, industrially relevant concentrations of substrate start at 100 g/L carbohydrate, which corresponds to approximately 150 g/L solids. To gain insight into the physiology of fermentation of high substrate concentrations, we studied the growth on, and utilization of high concentrations of crystalline cellulose varying from 50 to 100 g/L by C. thermocellum. Results: Using a defined medium, batch cultures of C. thermocellum achieved 93% conversion of cellulose (Avicel) initially present at 100 g/L. The maximum rate of substrate utilization increased with increasing substrate loading. During fermentation of 100 g/L cellulose, growth ceased when about half of the substrate had been solubilized. However, fermentation continued in an uncoupled mode until substrate utilization was almost complete. In addition to commonly reported fermentation products, amino acids - predominantly L-valine and L-alanine - were secreted at concentrations up to 7.5 g/L. Uncoupled metabolism was also accompanied by products not documented previously for C. thermocellum, including isobutanol, meso- and RR/SS-2,3-butanediol and trace amounts of 3-methyl-1-butanol, 2-methyl-1-butanol and 1-propanol. We hypothesize that C. thermocellummore »
- Authors:
-
- Dartmouth College, Hanover, NH (United States). Thayer School of Engineering; Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center (BESC)
- Mascoma Corp., Lebanon, NH (United States)
- Univ. of Wisconsin, Madison, WI (United States). Dept. of Bacteriology
- Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center (BESC); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Biosciences Division
- Delft Univ. of Technology (Netherlands)
- Dartmouth College, Hanover, NH (United States). Thayer School of Engineering; Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center (BESC); Mascoma Corp., Lebanon, NH (United States)
- Publication Date:
- Research Org.:
- Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center (BESC)
- Sponsoring Org.:
- USDOE Office of Science (SC)
- OSTI Identifier:
- 1163586
- Grant/Contract Number:
- AC05-00OR22725
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Biotechnology for Biofuels
- Additional Journal Information:
- Journal Volume: 7; Journal Issue: 1; Journal ID: ISSN 1754-6834
- Publisher:
- BioMed Central
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 09 BIOMASS FUELS; Clostridium thermocellum; Cellulose fermentation; Isobutanol; 2,3-butanediol; Amino acids; High solids; Fusel alcohols
Citation Formats
Holwerda, Evert K., Thorne, Philip G., Olson, Daniel G., Amador-Noguez, Daniel, Engle, Nancy L., Tschaplinski, Timothy J., van Dijken, Johannes P., and Lynd, Lee R. The exometabolome of Clostridium thermocellum reveals overflow metabolism at high cellulose loading. United States: N. p., 2014.
Web. doi:10.1186/s13068-014-0155-1.
Holwerda, Evert K., Thorne, Philip G., Olson, Daniel G., Amador-Noguez, Daniel, Engle, Nancy L., Tschaplinski, Timothy J., van Dijken, Johannes P., & Lynd, Lee R. The exometabolome of Clostridium thermocellum reveals overflow metabolism at high cellulose loading. United States. https://doi.org/10.1186/s13068-014-0155-1
Holwerda, Evert K., Thorne, Philip G., Olson, Daniel G., Amador-Noguez, Daniel, Engle, Nancy L., Tschaplinski, Timothy J., van Dijken, Johannes P., and Lynd, Lee R. Tue .
"The exometabolome of Clostridium thermocellum reveals overflow metabolism at high cellulose loading". United States. https://doi.org/10.1186/s13068-014-0155-1. https://www.osti.gov/servlets/purl/1163586.
@article{osti_1163586,
title = {The exometabolome of Clostridium thermocellum reveals overflow metabolism at high cellulose loading},
author = {Holwerda, Evert K. and Thorne, Philip G. and Olson, Daniel G. and Amador-Noguez, Daniel and Engle, Nancy L. and Tschaplinski, Timothy J. and van Dijken, Johannes P. and Lynd, Lee R.},
abstractNote = {Background: Clostridium thermocellum is a model thermophilic organism for the production of biofuels from lignocellulosic substrates. The majority of publications studying the physiology of this organism use substrate concentrations of ≤10 g/L. However, industrially relevant concentrations of substrate start at 100 g/L carbohydrate, which corresponds to approximately 150 g/L solids. To gain insight into the physiology of fermentation of high substrate concentrations, we studied the growth on, and utilization of high concentrations of crystalline cellulose varying from 50 to 100 g/L by C. thermocellum. Results: Using a defined medium, batch cultures of C. thermocellum achieved 93% conversion of cellulose (Avicel) initially present at 100 g/L. The maximum rate of substrate utilization increased with increasing substrate loading. During fermentation of 100 g/L cellulose, growth ceased when about half of the substrate had been solubilized. However, fermentation continued in an uncoupled mode until substrate utilization was almost complete. In addition to commonly reported fermentation products, amino acids - predominantly L-valine and L-alanine - were secreted at concentrations up to 7.5 g/L. Uncoupled metabolism was also accompanied by products not documented previously for C. thermocellum, including isobutanol, meso- and RR/SS-2,3-butanediol and trace amounts of 3-methyl-1-butanol, 2-methyl-1-butanol and 1-propanol. We hypothesize that C. thermocellum uses overflow metabolism to balance its metabolism around the pyruvate node in glycolysis. In conclusion: C. thermocellum is able to utilize industrially relevant concentrations of cellulose, up to 93 g/L. We report here one of the highest degrees of crystalline cellulose utilization observed thus far for a pure culture of C. thermocellum, the highest maximum substrate utilization rate and the highest amount of isobutanol produced by a wild-type organism.},
doi = {10.1186/s13068-014-0155-1},
journal = {Biotechnology for Biofuels},
number = 1,
volume = 7,
place = {United States},
year = {Tue Oct 21 00:00:00 EDT 2014},
month = {Tue Oct 21 00:00:00 EDT 2014}
}
Web of Science
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