title: N-carbamyl-L-amino acid amidohydrolase of Pseudomonas sp. strain NS671: purification and some properties of the enzyme expressed in Escherichia coli.
abstract: An N-carbamyl-L-amino acid amidohydrolase was purified from cells of Escherichia coli in which the gene for N-carbamyl-L-amino acid amidohydrolase of Pseudomonas sp. strain NS671 was expressed. The purified enzyme was homogeneous by the criterion of SDS-polyacrylamide gel electrophoresis. The results of gel filtration chromatography and SDS-polyacrylamide gel electrophoresis suggested that the enzyme was a dimeric protein with 45-kDa identical subunits. The enzyme required Mn2+ ion (above 1 mM) for the activity. The optimal pH and temperature were 7.5 and around 40 degrees C, respectively, with N-carbamyl-L-methionine as the substrate. The enzyme activity was inhibited by ATP and was lost completely with p-chloromercuribenzoate (1 mM). The enzyme was strictly L-specific and showed a broad substrate specificity for N-carbamyl-l-alpha-amino acids.
authors.affiliation: Odawara Research Center, Nippon Soda Co., Ltd., Kanagawa, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Ishikawa
authors.forename: K
authors.initials: K
authors.lastname: Watabe
authors.forename: Y
authors.initials: Y
authors.lastname: Mukohara
authors.forename: H
authors.initials: H
authors.lastname: Nakamura
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology; D000581:Amidohydrolases / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D002729:Chloromercuribenzoates / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D008345:Manganese / Q000494:pharmacology; D008715:Methionine / Q000378:metabolism; D011549:Pseudomonas; D011994:Recombinant Proteins / Q000096:biosynthesis; D013379:Substrate Specificity; D013696:Temperature; D020245:p-Chloromercuribenzoic Acid
chemical_list: D002729:Chloromercuribenzoates; D011994:Recombinant Proteins; D008345:Manganese; D020245:p-Chloromercuribenzoic Acid; D000255:Adenosine Triphosphate; D008715:Methionine; D000581:Amidohydrolases; C085345:1-carbamoyl-L-amino acid amidohydrolase