high level secretion of cellobiohydrolases by saccharomyces cerevisiae高水平的分泌cellobiohydrolases酿酒酵母.pdfVIP

Ilmén et al. Biotechnology for Biofuels 2011, 4:30 /content/4/1/30 RESEARCH Open Access High level secretion of cellobiohydrolases by Saccharomyces cerevisiae 1 2 3 3 3 3 1 Marja Ilmén , Riaan den Haan , Elena Brevnova , John McBride , Erin Wiswall , Allan Froehlich , Anu Koivula , 1 1 2 3 3 3 Sanni P Voutilainen , Matti Siika-aho , Daniël C la Grange , Naomi Thorngren , Simon Ahlgren , Mark Mellon , Kristen Deleault3, Vineet Rajgarhia3,4, Willem H van Zyl2 and Merja Penttilä1* Abstract Background: The main technological impediment to widespread utilization of lignocellulose for the production of fuels and chemicals is the lack of low-cost technologies to overcome its recalcitrance. Organisms that hydrolyze lignocellulose and produce a valuable product such as ethanol at a high rate and titer could significantly reduce the costs of biomass conversion technologies, and will allow separate conversion steps to be combined in a consolidated bioprocess (CBP). Development of Saccharomyces cerevisiae for CBP requires the high level secretion of cellulases, particularly cellobiohydrolases. Results: We expressed various cellobiohydrolases to identify enzymes that were efficiently secreted by S. cerevisiae. For enhanced cellulose hydrolysis, we engineered bimodular derivatives of a well secreted enzyme that naturally lacks the carbohydrate-binding module, and constructed strains expressing combinations of cbh1 and cbh2 genes. Though there was significant variability in the enzyme levels produced, up to approximately 0.3 g/L CBH1 and approximately 1 g/L CBH2 could be produced