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KMID : 0613820050150010118
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Journal of Life Science
2005 Volume.15 No. 1 p.118 ~ p.123
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Surface Display of Bacillus CGTase on the Cell of Saccharomyces cerevisiae
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Kim Hyun-Chul
Lim Chae-Kwon
Kim Byung-Woo
Jeon Seong-Soo
Nam Soo-Wan
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Abstract
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For the expression in Saccharomyces cerevisiae, Bacillus stearothermophilus cyclodextrin glucanotransferase gene (cgtS) in pCGTS (4.8 kb) was subcloned into the surface expression vector, pYD1 (GAL1 promoter). The constructed plasmid, pYDCGT (7.2 kb) was introduced into S. cerevisiae EBY100 cells, and then yeast transformants were selected on the synthetic defined media lacking tryptophan. The formation of cyclodextrin (CD) was confirmed with active staining of culture broth of transformant grown on starch medium. Enzymatic reaction products with respect to the culture time and the reaction time were examined by TLC analysis. The results indicated that the enzyme activity was exhibited after 12 h cultivation and CD was produced after 10 min of enzymatic reaction. When the surface-engineered yeast cells were cultured on galactose medium, maximum activities of CGTase were about 21.3 unit/l and 16.5 unit/l at 25℃ and 30℃, respectively. The plasmids stability showed about 80% even at 25℃ and 30℃.
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KEYWORD
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