Cellulase enzyme cocktails have potential value in the biorefining of renewable feedstock (such as crop residues, straws, and grasses) to biofuels and other bioproducts. Clostridium cellulovorans is a useful source of cellulase genes because it produces cellulase systems that degrade crystalline cellulose. The genes of three C. cellulovorans cellulases (endoglucanase, exoglucanase, and b-glucosidase) were cloned into the industrial yeast Saccharomyces cerevisiae. The cellulase genes were cloned individually, and also co-cloned, into S. cerevisiae. Active C. cellulovorans cellulase enzymes were constitutively expressed and secreted by the engineered yeast. Enzyme activity was detected using blue dye-labeled CM-cellulose and cello-oligosaccharide substrates.  The degradation of CMC in engineered yeast cultures was analyzed. PCR was used to confirm the presence of the three cellulase genes in engineered yeast clones. This cellulase enzyme complex is being studied for the digestion of cellulosic biomass to fermentable sugars.

Cellulase enzyme complex produced by engineered yeast

248th National Meeting (2014)

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