Nitrile-synthesizing enzyme: Gene cloning, overexpression and application for the production of useful compounds

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  • Kumano Takuto
    Institute of Applied Biochemistry, and Graduate School of Life and Environmental Sciences, University of Tsukuba
  • Takizawa Yuko
    Division of Applied Life Science, Graduate School of Agriculture, Kyoto University
  • Shimizu Sakayu
    Division of Applied Life Science, Graduate School of Agriculture, Kyoto University
  • Kobayashi Michihiko
    Institute of Applied Biochemistry, and Graduate School of Life and Environmental Sciences, University of Tsukuba

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Abstract

One of the nitrile-synthesizing enzymes, β-cyano-L-alanine synthase, catalyzes β-cyano-L-alanine (β-CNAla) from potassium cyanide and O-acetyl-L-serine or L-cysteine. We have identified this enzyme from Pseudomonas ovalis No. 111. In this study, we cloned the β-CNAla synthase gene and expressed it in Escherichia coli and Rhodococcus rhodochrous. Furthermore, we carried out co-expression of β-CNAla synthase with nitrilase or nitrile hydratases in order to synthesize aspartic acid and asparagine from KCN and O-acetyl-L-serine. This strategy can be used for the synthesis of labeled amino acids by using a carbon-labeled KCN as a substrate, resulting in an application for positron emission tomography.

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