Isolation, Analysis, and Expression of Lipase with Cephalosporin-C Deacetylation Activity from Staphylococcus sp.

  • Lee, Hyun-Woo (Department of Biological Sciences, Korea Advanced Institute of Science and Technology) ;
  • Ko, Jung-Youn (Department of Biological Sciences, Korea Advanced Institute of Science and Technology) ;
  • Kim, Woo-Jung (Department of Food Sciences, Sejong University) ;
  • Byun, Si-Myung (Department of Biological Sciences, Korea Advanced Institute of Science and Technology)
  • Received : 2001.01.26
  • Accepted : 2001.02.21
  • Published : 2001.05.31

Abstract

Lipase of Staphylococcus sp. was purified from the culture supernatant, and its molecular mass estimated to be 44 kDa by SDS-PAGE. Its optimum temperature and pH for the hydrolysis of p-nitrophenyl substrates was $28^{\circ}C$ and pH 8.5, respectively The gene encoding the lipase was cloned in Escherichia coli in the $NH_2$-teminally truncated form by using the shotgun method, and sequenced. The mature enzyme had a 49-93% amino acid sequence homology with other staphylococcal lipases. This lipase was used for the hydrolysis of the 3-O-acetate of cephalosporin-C to give an intermediate, deacetylated cephalosporin-C that is useful for further chemical elaborations.

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