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<em>N</em>-Carbamoyl-D-amino acid amidohydrolase from <em>Comamonas</em> sp. E222c. Purification and characterization.
- Source :
-
European Journal of Biochemistry . 3/15/93, Vol. 212 Issue 3, p685-691. 7p. - Publication Year :
- 1993
-
Abstract
- N-Carbamoyl-D-amino acid amidohydrolase was purified 119-fold, with 36% overall recovery from a cell-free extract of Comamonas sp. E222c. The purified enzyme was homogeneous as judged by SDS/PAGE. The relative molecular mass of the native enzyme was 120000 and that of the subunit was 40000. The purified enzyme hydrolyzed various N-carbamoyl-D-amino acids to Damino acids, ammonia and carbon dioxide. N-Carbamoyl-D-amino acids having hydrophobic groups served as good substrates for the enzyme. The Km and Vmax values for N-carbamoyl-D-phenylalanine were 19.7 mM and 13.1 units/mg, respectively, and those for N-carbamoyl-D-p-hydroxyphenylglycine were 13.1 mM and 0.56 units/mg, respectively. The enzyme strictly recognized the configuration of the substrate and only the D-enantiomer of the N-carbamoyl amino acid was hydrolyzed. The enzyme activity was not significantly affected by N-carbamoyl-L-amino acids and ammonia. The enzyme was sensitive to thiol reagents and did not require metal ions for its activity. The enzyme did not hydrolyze N-carbamoyl-β-alanine or N-carbamoyl-DL-aspartate suggesting that the enzyme is different from the N-carbamoylamide-hydrolyzing enzymes involved in the pyrimidine degradation pathway. The enzyme did not hydrolyze allantoin and allantoic acid, which are intermediates in purine degradation, N-carbamoylsarcosine and citrulline, suggesting that it is a novel N-carbamoylamide amidohydrolase. [ABSTRACT FROM AUTHOR]
- Subjects :
- *AMIDASES
*HYDROLASES
*AMINO acids
*ORGANIC acids
*ENZYMES
*CARBON dioxide
Subjects
Details
- Language :
- English
- ISSN :
- 00142956
- Volume :
- 212
- Issue :
- 3
- Database :
- Academic Search Index
- Journal :
- European Journal of Biochemistry
- Publication Type :
- Academic Journal
- Accession number :
- 13688768
- Full Text :
- https://doi.org/10.1111/j.1432-1033.1993.tb17706.x