1. N-Carbamyl-<scp>L</scp>-Amino Acid Amidohydrolase ofPseudomonassp. Strain NS671: Purification and Some Properties of the Enzyme Expressed inEscherichia coli
- Author
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Ken Watabe, Takahiro Ishikawa, Hiroaki Nakamura, and Yukuo Mukohara
- Subjects
p-Chloromercuribenzoic Acid ,Size-exclusion chromatography ,Biology ,medicine.disease_cause ,Applied Microbiology and Biotechnology ,Biochemistry ,Amidohydrolases ,Substrate Specificity ,Analytical Chemistry ,Adenosine Triphosphate ,Methionine ,Pseudomonas ,medicine ,Molecular Biology ,Escherichia coli ,chemistry.chemical_classification ,Gel electrophoresis ,Manganese ,Chromatography ,Amidohydrolase ,Organic Chemistry ,Temperature ,Substrate (chemistry) ,General Medicine ,Hydrogen-Ion Concentration ,Recombinant Proteins ,Enzyme assay ,Amino acid ,Enzyme ,chemistry ,biology.protein ,Chloromercuribenzoates ,Biotechnology - 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. more...
- Published
- 1996
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