N-Carbamyl-L-amino acid amidohydrolase of Pseudomonas sp. strain NS671: Purification and some properties of the enzyme expressed in Escherichia coli

Article abstract of DOI:10.1271/bbb.60.612

An N-carbamyl-L-amino acid amidohydrotase 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°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-α-amino acids.

Full text of DOI:10.1271/bbb.60.612

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N-Carbamyl-L-Amino Acid Amidohydrolase of
Pseudomonas sp. Strain NS671: Purification and
Some Properties of the Enzyme Expressed in
Escherichia coli
Takahiro Ishikawaa, Ken Watabea, Yukuo Mukoharaa & Hiroaki Nakamuraa
a Odawara Research Center, Nippon Soda Co., Ltd., 345 Takada, Odawara, Kanagawa
250-02, Japan
Published online: 12 Jun 2014.
To cite this article: Takahiro Ishikawa, Ken Watabe, Yukuo Mukohara & Hiroaki Nakamura (1996) N-Carbamyl-L-Amino
Acid Amidohydrolase of Pseudomonas sp. Strain NS671: Purification and Some Properties of the Enzyme Expressed in
Escherichia coli, Bioscience, Biotechnology, and Biochemistry, 60:4, 612-615, DOI: 10.1271/bbb.60.612
To link to this article: http://dx.doi.org/10.1271/bbb.60.612
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