Expression of a thermostable cellulase gene from a thermophilic anaerobe in Saccharomyces cerevisiae |
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| Affiliation: | 1. Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China;2. Lab of Brewing Science and Technology, School of Biotechnology, Jiangnan University, Wuxi 214122, China;3. School of Biotechnology, Jiangnan University, Wuxi 214122, China;1. Matis, Reykjavik, Iceland;2. Faculty of Life and Environmental Sciences, University of Iceland, Reykjavik, Iceland;3. Biotechnology, Dept of Chemistry, Lund University, Lund, Sweden;4. SINTEF Industry, Trondheim, Norway;5. NOBIPOL, Dept of Biotechnology and Food Science, NTNU Norwegian University of Science and Technology, Trondheim, Norway;6. Department of Chemical Biology and Drug Discovery, Utrecht Institute for Pharmaceutical Sciences, and Bijvoet Center for Biomolecular Research, Utrecht University, Utrecht, Netherlands |
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| Abstract: | A cellulase gene from a thermophilic anaerobe was recloned in the yeast Saccharomyces cerevisiae. The maximum level of the gene expression in the recombinant yeast was 4.4 times higher than that in the Escherichia coli transformant harboring the same plasmid. Cellulase activity was observed only within the yeast cells. To compare the enzymatic properties of cellulase produced by the yeast and E. coli transformants, cellulases were purified to homogeneous state by only three purification steps of heat treatment, and cellulose affinity and ion exchange chromatographies. The molecular weights of the enzymes produced by the yeast and E. coli were 3.8 × 104 and 4.0 × 104, respectively by SDS-polyacrylamide gel electrophoresis. Neither of the enzymes was glycosylated. Although the molecular weights were slightly different, enzymatic properties and thermostability were almost indistinguishable between the enzymes produced by the yeast and E. coli transformants. |
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