Gene Cloning and Expression of Cephalosporin - C Deacetylase from Bacillus sp . KCCM10143

• Published in: Journal of microbiology and biotechnology Vol. 10; no. 2; pp. 221 - 226
• Main Authors: Duk Ho Choi, Yong Duk Kim, Il Sun Chung, Sang Hun Lee, 강상모, Sang Mo Kang, Tae Jong Kwon, Kum Soo Han
• Format: Journal Article
• Language: Korean
• Published: 한국미생물생명공학회 01.04.2000
• Subjects: 7-aminocephalosporanic acid (7-ACA); deacetylase; Cephalosporin; Bacillus
• ISSN: 1017-7825; 1738-8872

Cephalosporin-C deacetylase (CAH) catalyzes the deacetylation of cephalosporin derivatives. A novel gene encoding the CAH from Bacillus sp. KCCM10143 was cloned and sepuenced. The uncleotide sequence contained an open reading frame encoding a polypeptide consisting of 217 amino acids and a molecular weight of 24 kDa 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 from of the CAH protein was expressed in the soluble fraction obtained after cell disruption. in fermentation using a 5-1 jar fementer, the transformant E. coli JM109 (pDST654) produced 4.12 U of CAH per ml of culture during 16 h of incubation.