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

Suresh Poudel, Richard J. Giannone, Miguel Rodriguez, Babu Raman, Madhavi Z. Martin, Nancy L. Engle, Jonathan R. Mielenz, Intawat Nookaew, Steven D. Brown, Timothy J. Tschaplinski, David Ussery, Robert L. Hettich

Research output: Contribution to journalArticlepeer-review

27 Scopus citations

Abstract

Background: Clostridium thermocellum is capable of solubilizing and converting lignocellulosic biomass into ethanol. Although much of the work-to-date has centered on characterizing this microbe's growth on model cellulosic substrates, such as cellobiose, Avicel, or filter paper, it is vitally important to understand its metabolism on 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. Results: 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 undoubtedly a response to the gradual accumulation of lignocellulose-derived inhibitory compounds as the organism deconstructs the switchgrass to access the embedded cellulose. Corroborating the metabolomics data, proteomic analysis revealed a corresponding time-dependent increase in various enzymes, including those involved in the interconversion of branched amino acids valine, leucine, and isoleucine to iso- and anteiso-fatty acid precursors. Additionally, the metabolic accumulation of hemicellulose-derived sugars and sugar alcohols concomitant with increased abundance of enzymes involved in C5 sugar metabolism/pentose phosphate pathway indicates that C. thermocellum shifts glycolytic intermediates to alternate pathways to modulate overall carbon flux in response to C5 sugar metabolites that increase during lignocellulose deconstruction. Conclusions: Integrated omic platforms provided complementary systems biological information that highlight C. thermocellum's specific response to cytotoxic inhibitors released during the deconstruction and utilization of switchgrass. These additional viewpoints allowed us to fully realize the level to which the organism adapts to an increasingly challenging culture environment - information that will prove critical to C. thermocellum's industrial efficacy.

Original languageEnglish
Article number14
JournalBiotechnology for Biofuels
Volume10
Issue number1
DOIs
StatePublished - Jan 10 2017

Funding

The authors would like to acknowledge ORNL staff members Zamin K. Yang for help with RNA extractions/QC, and Lezlee Dice for DNA microarray work. Additional acknowledgement is given to ORNL researchers Adam Guss and James Elkins for helpful technical comments during the preparation of this manuscript. This study was funded by the BioEnergy Science Center, a US Department of Energy Bioenergy Research Center supported by the Office of Biological and Environmental Research in the DOE Office of Science. Oak Ridge National Laboratory is managed by University of Tennessee-Battelle LLC for the Department of Energy under contract DOE-AC05-00OR22725. This study was funded by the BioEnergy Science Center, a US Department of Energy Bioenergy Research Center supported by the Office of Biological and Environmental Research in the US DOE Office of Science.

Keywords

  • Biofuel
  • Cellulosome
  • Clostridium thermocellum
  • Ethanol
  • Lignocellulosic
  • Mass spectrometry
  • Metabolomics
  • Proteomics
  • Switchgrass
  • Transcriptomics

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