Abstract
BackgroundClostridium 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.ResultsThe 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.ConclusionsIntegrated 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.
Highlights
Clostridium thermocellum is capable of solubilizing and converting lignocellulosic biomass into ethanol
Acetic acid can be generated through deacetylation of hemicelluloses, [27] and the relative contributions from the pretreated biomass or bacteria cannot be differentiated in this study
Our results indicate that C. thermocellum increases the production of long-chain, branched iso- and anteisofatty acids over time, perhaps in response to the adverse effect imposed by lignocellulose-derived inhibitors and fermentation end products
Summary
Clostridium thermocellum is capable of solubilizing and converting lignocellulosic biomass into ethanol. 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. This Gram-positive bacterium can be found in natural environments where cellulose degradation actively occurs (e.g., compost piles) It produces large extracellular enzyme complexes called cellulosomes that are predominantly tethered to the cell surface but can exist as free entities, enabling the efficient solubilization and deconstruction of lignocellulose to simpler sugars [6, 7]. Paired with the organism’s innate ability to ferment sugar to ethanol, the presence of cellulosomes makes C. thermocellum, an ideal candidate for consolidated bioprocessing (CBP), a “one-pot” industrial process whereby lignocellulosic biomass is converted directly into biofuel [8]
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