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KMID : 0545120000100020221
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Journal of Microbiology and Biotechnology
2000 Volume.10 No. 2 p.221 ~ p.226
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Gene Cloning and Expression of Cephalosporin-C Deacetylase from Bacillus sp.KCCM10143
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Choi, Duk Ho
Kim, Yong Duk/Chung, Il Sun/Lee, Sang Hun/Kang, Sang Mo/Kwon, Tae Jong/Han, Kum Soo
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Abstract
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Cephalosporin-C deacetylase (CAH) catalyzes the deacetylation of cephalosporin derivatives. A novel gene encoding the CAH from Bacillus sp. KCCM10143 was cloned and sequenced. The nucleotide sequence contained an open reading frame encoding a polypeptide consisting of 217 amino acids and a molecular weight of 24kDa which was in good agreement with the value obtained by sodium dodecylsulfate-polyacrylamide gel electrophoresis. An expression plasmid was constructed by inserting the CAH gene into the region of the pTrc99A expression vector. An active form of the CAH protein was expressed in the soluble fraction obtained after cell disruption. In fermentation using a 5-1 jar fermenter, the transformant E. coli JM 109 (pDST654) produced 4.12 U of CAH per ml of culture during 16h of incubation.
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