Abstract
Our purpose was to identify the sequence of omega-amidase, which hydrolyses the amide group of alpha-ketoglutaramate, a product formed by glutamine transaminases. In the Bacillus subtilis genome, the gene encoding a glutamine transaminase (mtnV) is flanked by a gene encoding a putative 'carbon-nitrogen hydrolase'. The closest mammalian homolog of this putative bacterial omega-amidase is 'nitrilase 2', whose size and amino acid composition were in good agreement with those reported for purified rat liver omega-amidase. Mouse nitrilase 2 was expressed in Escherichia coli, purified and shown to catalyse the hydrolysis of alpha-ketoglutaramate and other known substrates of omega-amidase. No such activity was observed with mouse nitrilase 1. We conclude that mammalian nitrilase 2 is omega-amidase.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amidohydrolases / chemistry
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Amidohydrolases / genetics
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Amidohydrolases / metabolism*
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Amino Acid Sequence
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Aminohydrolases / chemistry
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Aminohydrolases / genetics
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Aminohydrolases / metabolism*
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Animals
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Bacillus subtilis / enzymology
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Bacillus subtilis / genetics
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Bacterial Proteins / chemistry
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Bacterial Proteins / genetics
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Bacterial Proteins / metabolism*
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Blotting, Western
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Cell Line
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Computational Biology
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Databases, Genetic
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Escherichia coli / genetics
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Escherichia coli / metabolism
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Genome, Bacterial / genetics
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Humans
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Ketoglutaric Acids / metabolism
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Mice
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Models, Genetic
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Molecular Sequence Data
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Sequence Homology, Amino Acid
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Transaminases / chemistry
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Transaminases / genetics
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Transaminases / metabolism*
Substances
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Bacterial Proteins
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Ketoglutaric Acids
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alpha-ketoglutaramate
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Transaminases
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glutamine-pyruvate aminotransferase
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Amidohydrolases
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omega-amidase
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Aminohydrolases