Cloning and expression of D-amino acid oxidase from Trigonopsis variabilis for cephalosporin C biotransformation = Cephalosporin C의 생변환을 위한 Trigonopsis variabilis의 D-amino acid Oxidase 유전자의 클로닝 및 발현

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Title
Cloning and expression of D-amino acid oxidase from Trigonopsis variabilis for cephalosporin C biotransformation = Cephalosporin C의 생변환을 위한 Trigonopsis variabilis의 D-amino acid Oxidase 유전자의 클로닝 및 발현
Author(s)
Jin Hyung Lee; Tae Whan Chung; Sung Gon Kwak; Dong Joon Lim; Cheol Sik Yoon; Sang Ki Rhee; Yong Ho Khang
Bibliographic Citation
Korean Journal of Biotechnology and Bioengineering, vol. 10, no. 3, pp. 264-270
Publication Year
1995
Abstract
Trigonopsis variabilis is a strong producer of D-amino acid oxidase that can transform cephalosporin C(ceph C) to -keto-adipyl-7-aminocephalosporanic acid(AKA-7ACA). Polymerase chain reaction (PCR) was applied to isolate the D-AAO gene from T. variabilis. To clone the PCR fragment, four different methods were examined using enzymatic reactions of Taq DNA polymerase, Klenow, T4 DNA polymerase I, Alkaline phosphatase Calf Intestinal, and T4 kinase. Ligation of phosphorylated blunt-end PCR fragment and dephosphorylated blunt-end of pUC18 plasmid yielded the best cloning efficiency One of recombinant E. coli transformants showed D-AAO activity against ceph C in both cell extracts and permeabilized cells.
ISSN
1225-7117
Publisher
Korea Soc-Assoc-Inst
Type
Article
Appears in Collections:
1. Journal Articles > Journal Articles
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