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KMID : 1007519950040040290
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Food Science and Biotechnology
1995 Volume.4 No. 4 p.290 ~ p.295
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Expression of Cyclodextrin Glucanotransferase from Bacillus macerans in Recombinant Escherichia coli
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SEO, JIN HO
Park, Yong Cheol/Kim, Chang Sup/Han, Nam Soo
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Abstract
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A gene encoding cyclodextrin glucanotransferase (CGTase) of Bacillus macerans was expressed in recombinant E. coli BL 21(DE3) containing plasmid pTCGT1. The cgt gene coding for CGTase was under the control of the T7 promoter so that its expression was able to be induced by isopropyl-β-D-thiogalactopyranoside (IPTG). B. macerans CGTase produced α-cyclodextrin (α-CD) from starch as a major compound with a 67% selectivity. Induction of the T7 promoter by IPTG at the late exponential phase resulted in the highest CGTase activity of 1.0 U/㎖. BL21(DE3)pLysE was chosen as the best host for the T7 promoter system as the recombinant strain BL21(DE3) pLysS containing plasmid pTCGT1 yielded a CGTase activity of 3.8 U/㎖, which was higher than the CGTase levels obtained from the other systems such as BL21(DE3) and BL21(DE3)pLysS. An SDS-PAGE analysis authorstrongly suggested that the CGTase was accumulated within the cell as a form of inclusion body.
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KEYWORD
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