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KMID : 0545119990090020230
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Journal of Microbiology and Biotechnology
1999 Volume.9 No. 2 p.230 ~ p.233
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Expression of Bacillus macerans Cyclodextrin Glucanotransferase in Bacillus subtilis
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Kim, Chang Sup
Han, Nam Soo/Kweon, Dae Hyuk/Seo, Jin Ho
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Abstract
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A plasmid vector was constructed for the expression and secretion of Bacillus macerans cyclodextrin glucanotransferase (CGTase) in Bacillus subtilis. The vector, pUBACGT, was composed of the ribosome-binding sequence, signal sequence, and cgt gene from B. macerans under the control of amyR2, the promoter of amyE gene coding for α-amylase froth B. subtilis var. natto. Bacillus subtilis LKS88, a mutant strain lacking genes for an amylase and two proteases, was used as a host for the transformation of the plasmid vector. The transformants were selected on kanamycin-containing Luria-Bertani plates. The starch hydrolyzing activity was observed on the starch-containing plates by the iodine method and cyclodextrin-forming activity was detected in the culture medium. A SDS-PAGE analysis showed that most of the expressed CGTase in the recombinant B. subtilis was secreted into the medium at a high expression level.
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