title: Nitrile-synthesizing enzyme: Gene cloning, overexpression and application for the production of useful compounds.
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.
authors.affiliation: Institute of Applied Biochemistry, and Graduate School of Life and Environmental Sciences, University of Tsukuba.
authors.forename: Takuto
authors.initials: T
authors.lastname: Kumano
authors.forename: Yuko
authors.initials: Y
authors.lastname: Takizawa
authors.forename: Sakayu
authors.initials: S
authors.lastname: Shimizu
authors.forename: Michihiko
authors.initials: M
authors.lastname: Kobayashi
mesh_terms: D000409:Alanine / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism; D000596:Amino Acids / Q000737:chemistry; D000619:Aminohydrolases / Q000235:genetics; D001216:Asparagine / Q000096:biosynthesis; D001224:Aspartic Acid / Q000096:biosynthesis; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D006836:Hydro-Lyases / Q000235:genetics / Q000378:metabolism; D008190:Lyases / Q000235:genetics* / Q000378:metabolism*; D009570:Nitriles / Q000378:metabolism*; D049268:Positron-Emission Tomography; D011190:Potassium Cyanide / Q000378:metabolism; D011549:Pseudomonas / Q000201:enzymology* / Q000235:genetics; D012240:Rhodococcus / Q000235:genetics* / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D000596:Amino Acids; D009570:Nitriles; D001224:Aspartic Acid; D001216:Asparagine; C004631:3-cyanoalanine; D000619:Aminohydrolases; C021867:nitrilase; D008190:Lyases; D006836:Hydro-Lyases; C048831:nitrile hydratase; C019955:beta-cyanoalanine synthase; D011190:Potassium Cyanide; D000409:Alanine