Journal of Microbiology and Biotechnology

  • 제18권1호
  • /
  • Pages.48-54
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  • 2008
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  • 1017-7825(pISSN)
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  • 1738-8872(eISSN)
한국미생물·생명공학회 (The Korean Society for Microbiology and Biotechnology)
권호 표지

Identification of ${\omega}$-Aminotransferase from Caulobacter crescentus and Sitedirected Mutagenesis to Broaden Substrate Specificity

  • Hwang, Bum-Yeol (School of Chemical Engineering, and Institute for Molecular Biology and Genetics, Seoul National University) ;
  • Ko, Seung-Hyun (School of Chemical Engineering, and Institute for Molecular Biology and Genetics, Seoul National University) ;
  • Park, Hyung-Yeon (Division of Chemistry, Inha University) ;
  • Seo, Joo-Hyun (School of Chemical Engineering, and Institute for Molecular Biology and Genetics, Seoul National University) ;
  • Lee, Bon-Su (Division of Chemistry, Inha University) ;
  • Kim, Byung-Gee (School of Chemical Engineering, and Institute for Molecular Biology and Genetics, Seoul National University)
  • 발행 : 2008.01.31
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초록

A putative ${\omega}$-aminotransferase gene, cc3143 (aptA), from Caulobacter crescentus was screened by bioinformatical tools and overexpressed in E. coli, and the substrate specificity of the ${\omega}$-aminotransferase was investigated. AptA showed high activity for short-chain ${\beta}$-amino acids. It showed the highest activity for 3-amino-n-butyric acid. It showed higher activity toward aromatic amines than aliphatic amines. The 3D model of the ${\omega}$-aminotransferase was constructed by homology modeling using a dialkylglycine decarboxylase (PDB ID: 1DGE) as a template. Then, the ${\omega}$-aminotransferase was rationally redesigned to increase the activity for 3-amino-3-phenylpropionic acid. The mutants N285A and V227G increased the relative activity for 3-amino-3-phenylpropionic acid to 3-amino-n-butyric acid by 11-fold and 3-fold, respectively, over that of wild type.

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