Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Molecular Characterization of the Genes Encoding Acetoacetyl-Coenzyme A Transferase from Serratia marcescens KCTC 2172

  • Yoo, Ju-Soon (Division of Biotechnology, Faculty of Natural Resources and Life Sciences, Dong-A University) ;
  • Kim, Hae-Sun (Division of Biotechnology, Faculty of Natural Resources and Life Sciences, Dong-A University) ;
  • Lee, Young-Choon (Division of Biotechnology, Faculty of Natural Resources and Life Sciences, Dong-A University) ;
  • Chung, Soo-Yeol (Department of Food Science and Nutrition, Donju College) ;
  • Choi, Yong-Lark (Division of Biotechnology, Faculty of Natural Resources and Life Sciences, Dong-A University)
  • Published : 2001.10.01
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Abstract

A DNA fragment, pCKB13, containing two genes encoding Coenzyme a transferase, was isolated from a genomic DNA library of S. marcescens KCTC 2172. The complete nucleotide sequence of the 2,081-bp BamHI fragment on pCKB13 was determined. Sequencing of the fragment led to the identification of two open reading frames showing high homology with two Coenzyme A (CoA) transferases, Acetoacetyl-CoA transferase (Acot) and Succinyl-CoA transferase (Scot), enzymes catalyzing the reversible transfer of CoA from one carboxylic acid to another. The enzyme activity of Coenzyme A transferase increased after introducing the multicopy of the cloned gene in E. coli. The recombinant protein, overexpressed by multicopy and induction with IPTG, was a polypeptide of 42 kDa, as confirmed by SDS-PAGE. The protein was purified to homogeneity through three sequential chromatographic procedures including ion-exchanged DEAE-sepharose, CM-sepharose, and Mono Q.

Keywords

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