A nitrilase from a metagenomic library acts regioselectively on aliphatic dinitriles.
Several novel nitrilases were selected from metagenomic libraries using cinnamonitrile and a mixture of six different nitriles as substrates. The nitrilase gene nit1 was expressed in Escherichia coli and the resulting protein was further examined concerning its biochemical properties. Nit1 turned out to be an aliphatic nitrilase favoring dinitriles over mononitriles. Stereochemical analysis revealed that Nit1 converted the dinitrile 2-methylglutaronitrile regioselectively. Hydrolysis at the ω-nitrile group of a dinitrile, such as catalyzed by Nit1, leads to ω-cyanocarboxylic acids, which are important precursors for chemical and pharmaceutical products. Nit1 metabolized 2-methylglutaronitrile to the corresponding ω-cyanocarboxylic acid 4-cyanopentanoic acid can be used for the production of the fine chemical 1,5-dimethyl-2-piperidone.
Bayer
Sally
S
Junior Research Group White Biotechnology, Institute of Biochemistry, Faculty of Biosciences, Pharmacy and Psychology, University of Leipzig, Deutscher Platz 5b, 04103 Leipzig, Germany.
Birkemeyer
Claudia
C
Ballschmiter
Meike
M
D000595:Amino Acid Sequence; D000619:Aminohydrolases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001419:Bacteria / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D007700:Kinetics; D056186:Metagenomics; D008969:Molecular Sequence Data; D009570:Nitriles / Q000737:chemistry* / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D012988:Soil Microbiology; D013379:Substrate Specificity; D014871:Water Microbiology
D001426:Bacterial Proteins; D009570:Nitriles; D011994:Recombinant Proteins; D000619:Aminohydrolases; C076876:aliphatic nitrilase