Proteomic profile changes in membranes of ethanol-tolerant <Emphasis Type="Italic">Clostridium thermocellum</Emphasis> |
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Authors: | Taufika Islam Williams Jennifer C Combs Bert C Lynn Herbert J Strobel |
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Institution: | (1) Department of Chemistry, University of Kentucky, Lexington, KY 40546, USA;(2) Department of Animal and Food Sciences, University of Kentucky, 212 Garrigus Building, Lexington, KY 40546, USA;(3) Present address: Department of Chemistry, North Carolina State University, Raleigh, NC, 27695, USA |
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Abstract: | Clostridium thermocellum, a cellulolytic, thermophilic anaerobe, has potential for commercial exploitation in converting fibrous biomass to ethanol.
However, ethanol concentrations above 1% (w/v) are inhibitory to growth and fermentation, and this limits industrial application of the organism. Recent work with ethanol-adapted
strains suggested that protein changes occurred during ethanol adaptation, particularly in the membrane proteome. A two-stage
Bicine-doubled sodium dodecyl sulfate-polyacrylamide gel electrophoresis protocol was designed to separate membrane proteins
and circumvent problems associated with membrane protein analysis using traditional gel-based proteomics approaches. Wild-type
and ethanol-adapted C. thermocellum membranes displayed similar spot diversity and approximately 60% of proteins identified from purified membrane fractions
were observed to be differentially expressed in the two strains. A majority (73%) of differentially expressed proteins were
down-regulated in the ethanol-adapted strain. Based on putative identifications, a significant proportion of these down-regulated
proteins were involved with carbohydrate transport and metabolism. Approximately one-third of the up-regulated proteins in
the ethanol-adapted species were associated with chemotaxis and signal transduction. Overall, the results suggested that membrane-associated
proteins in the ethanol-adapted strain are either being synthesized in lower quantities or not properly incorporated into
the cell membrane. |
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Keywords: | Clostridium thermocellum Ethanol tolerance Proteomics |
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