Abstract
The process of converting lignocellulosic biomass to ethanol via fermentation depends on developing economic sources of cellulases.Trichoderma reesei cellobiohydrolase (CBH) I is a key enzyme in the fungal cellulase system; however, specific process application requirements make modification of the enzyme by site-directed mutagenesis (SDM) an attractive goal. To undertake SDM investigations, an efficient, cellulase-free host is required. To test the potential ofEscherichia coli as a host, T.reesei CBH I cDNA was expressed inE. coli strain GI 724 as a C-terminal fusion to thermostable thioredoxin protein. Full-length expression of CBH I was subsequently verified by molecular weight, Western blot analysis, and activity on soluble substrates.
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Laymon, R.A., Adney, W.S., Mohagheghi, A. et al. Cloning and expression of full-lengthTrichoderma reesi cellobiohydrolase I cDNAs inEscherichia coli . Appl Biochem Biotechnol 57, 389–397 (1996). https://doi.org/10.1007/BF02941718
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DOI: https://doi.org/10.1007/BF02941718