Development of an arming yeast strain for efficient utilization of starch by co-display of sequential amylolytic enzymes on the cell surface |
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Authors: | T. Murai M. Ueda Y. Shibasaki N. Kamasawa M. Osumi T. Imanaka A. Tanaka |
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Affiliation: | (1) Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Yoshida, Sakyo-ku, Kyoto 606-8501, Japan Tel.: +81-75-753-5524 Fax: +81-75-753-5534, JP;(2) Department of Chemical and Biological Sciences, Faculty of Science, Japan Women's University, 2-8-1 Mejiro-dai, Bunkyo-ku, Tokyo 112-8681, Japan, JP |
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Abstract: | The construction of a whole-cell biocatalyst with its sequential reaction has been performed by the genetic immobilization of two amylolytic enzymes on the yeast cell surface. A recombinant strain of Saccharomyces cerevisiae that displays glucoamylase and α-amylase on its cell surface was constructed and its starch-utilizing ability was evaluated. The gene encoding Rhizopus oryzae glucoamylase, with its own secretion signal peptide, and a truncated fragment of the α-amylase gene from Bacillus stearothermophilus with the prepro secretion signal sequence of the yeast α factor, respectively, were fused with the gene encoding the C-terminal half of the yeast α-agglutinin. The constructed fusion genes were introduced into the different loci of chromosomes of S. cerevisiae and expressed under the control of the glyceraldehyde-3-phosphate dehydrogenase promoter. The glucoamylase and α-amylase activities were not detected in the culture medium, but in the cell pellet fraction. The transformant strain co-displaying glucoamylase and α-amylase could grow faster on starch as the sole carbon source than the transformant strain displaying only glucoamylase. Received: 16 June 1998 / Received last revision: 21 August 1998 / Accepted: 3 September 1998 |
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