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Title: Integrated omics analyses reveal the details of metabolic adaptation of Clostridium thermocellum to lignocellulose-derived growth inhibitors released during the deconstruction of switchgrass

Abstract

Clostridium thermocellum is capable of solubilizing and converting lignocellulosic biomass into ethanol. Though much of the work-to-date has centered on characterizing the organism s metabolism during growth on model cellulosic substrates, such as cellobiose, Avicel, or filter paper, it is vitally important to understand it metabolizes more complex, lignocellulosic substrates to identify relevant industrial bottlenecks that could undermine efficient biofuel production. To this end, we have examined a time course progression of C. thermocellum grown on switchgrass to assess the metabolic and protein changes that occur during the conversion of plant biomass to ethanol. The most striking feature of the metabolome was the observed accumulation of long-chain, branched fatty acids over time, implying an adaptive restructuring of C. thermocellum s cellular membrane as the culture progresses. This is likely a response to the gradual build-up of lignocellulose-derived inhibitory compounds detected as the organism deconstructs the switchgrass to access the embedded cellulose and includes 4-hydroxybenzoic acid, vanillic acid, ferulic acid, p-coumaric acid and vanillin. Corroborating the metabolomics data, proteomic analysis revealed a corresponding time-dependent increase in enzymes involved in the interconversion of branched amino acids valine, leucine and isoleucine to iso- and anteiso-fatty acid precursors. Furthermore, the metabolic accumulation of hemicellulose-derivedmore » sugars and sugar-alcohols concomitant with increased abundance of enzymes involved in C5 sugar metabolism / the pentose phosphate pathway, indicate that C. thermocellum either shifts glycolytic intermediates to alternate pathways to modulate overall carbon flux or is simply a response to C5 sugar metabolite pools that build during lignocellulose deconstruction.« less

Authors:
; ; ; ; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1618664
Alternate Identifier(s):
OSTI ID: 1340445
Grant/Contract Number:  
ORNL-BRC; AC05-00OR22725
Resource Type:
Journal Article: 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; Clostridium thermocellum; Switchgrass; Lignocellulosic; Biofuel; Ethanol; Mass spectrometry; Proteomics; Metabolomics; Transcriptomics; Cellulosome

Citation Formats

Poudel, Suresh, Giannone, Richard J., Rodriguez, Jr., Miguel, Raman, Babu, Martin, Madhavi Z., Engle, Nancy L., Mielenz, Jonathan R., Nookaew, Intawat, Brown, Steven D., Tschaplinski, Timothy J., Ussery, David, and Hettich, Robert L. Integrated omics analyses reveal the details of metabolic adaptation of Clostridium thermocellum to lignocellulose-derived growth inhibitors released during the deconstruction of switchgrass. Netherlands: N. p., 2017. Web. doi:10.1186/s13068-016-0697-5.
Poudel, Suresh, Giannone, Richard J., Rodriguez, Jr., Miguel, Raman, Babu, Martin, Madhavi Z., Engle, Nancy L., Mielenz, Jonathan R., Nookaew, Intawat, Brown, Steven D., Tschaplinski, Timothy J., Ussery, David, & Hettich, Robert L. Integrated omics analyses reveal the details of metabolic adaptation of Clostridium thermocellum to lignocellulose-derived growth inhibitors released during the deconstruction of switchgrass. Netherlands. doi:10.1186/s13068-016-0697-5.
Poudel, Suresh, Giannone, Richard J., Rodriguez, Jr., Miguel, Raman, Babu, Martin, Madhavi Z., Engle, Nancy L., Mielenz, Jonathan R., Nookaew, Intawat, Brown, Steven D., Tschaplinski, Timothy J., Ussery, David, and Hettich, Robert L. Tue . "Integrated omics analyses reveal the details of metabolic adaptation of Clostridium thermocellum to lignocellulose-derived growth inhibitors released during the deconstruction of switchgrass". Netherlands. doi:10.1186/s13068-016-0697-5.
@article{osti_1618664,
title = {Integrated omics analyses reveal the details of metabolic adaptation of Clostridium thermocellum to lignocellulose-derived growth inhibitors released during the deconstruction of switchgrass},
author = {Poudel, Suresh and Giannone, Richard J. and Rodriguez, Jr., Miguel and Raman, Babu and Martin, Madhavi Z. and Engle, Nancy L. and Mielenz, Jonathan R. and Nookaew, Intawat and Brown, Steven D. and Tschaplinski, Timothy J. and Ussery, David and Hettich, Robert L.},
abstractNote = {Clostridium thermocellum is capable of solubilizing and converting lignocellulosic biomass into ethanol. Though much of the work-to-date has centered on characterizing the organism s metabolism during growth on model cellulosic substrates, such as cellobiose, Avicel, or filter paper, it is vitally important to understand it metabolizes more complex, lignocellulosic substrates to identify relevant industrial bottlenecks that could undermine efficient biofuel production. To this end, we have examined a time course progression of C. thermocellum grown on switchgrass to assess the metabolic and protein changes that occur during the conversion of plant biomass to ethanol. The most striking feature of the metabolome was the observed accumulation of long-chain, branched fatty acids over time, implying an adaptive restructuring of C. thermocellum s cellular membrane as the culture progresses. This is likely a response to the gradual build-up of lignocellulose-derived inhibitory compounds detected as the organism deconstructs the switchgrass to access the embedded cellulose and includes 4-hydroxybenzoic acid, vanillic acid, ferulic acid, p-coumaric acid and vanillin. Corroborating the metabolomics data, proteomic analysis revealed a corresponding time-dependent increase in enzymes involved in the interconversion of branched amino acids valine, leucine and isoleucine to iso- and anteiso-fatty acid precursors. Furthermore, the metabolic accumulation of hemicellulose-derived sugars and sugar-alcohols concomitant with increased abundance of enzymes involved in C5 sugar metabolism / the pentose phosphate pathway, indicate that C. thermocellum either shifts glycolytic intermediates to alternate pathways to modulate overall carbon flux or is simply a response to C5 sugar metabolite pools that build during lignocellulose deconstruction.},
doi = {10.1186/s13068-016-0697-5},
journal = {Biotechnology for Biofuels},
issn = {1754-6834},
number = 1,
volume = 10,
place = {Netherlands},
year = {2017},
month = {1}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record at 10.1186/s13068-016-0697-5

Citation Metrics:
Cited by: 8 works
Citation information provided by
Web of Science

Figures / Tables:

Fig. 1 Fig. 1: Concentration (g/l) of acetic acid and ethanol (data reported as the average from triplicate fermentations on pretreated switch-grass). Arrows indicate the sampling points used for mass spectrometry (MS), metabolomics (Mtbl), and microarray (MA)

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      Figures/Tables have been extracted from DOE-funded journal article accepted manuscripts.