The purification and characterization of a novel D(-)-specific carbamoylase enzyme from an Agrobacterium sp.

被引：35

作者：

Louwrier, A ^{[1
]}

Knowles, CJ ^{[1
]}

机构：

[1] UNIV KENT,BIOL LAB,CANTERBURY,KENT,ENGLAND

来源：

ENZYME AND MICROBIAL TECHNOLOGY | 1996年 / 19卷 / 08期

关键词：

carbamoylase; hydantoinase; N-carbamoyl hydroxyphenylglycine; hydroxyphenylglycine; hydroxyphenyl hydantoin; D-AMINO ACIDS; CORRESPONDING 5-SUBSTITUTED HYDANTOINS; D; L-5-MONOSUBSTITUTED HYDANTOINS; (+)-1-PHENYLETHANESULFONIC ACID; BIOTECHNOLOGICAL PRODUCTION; ASYMMETRIC TRANSFORMATION; PARA-HYDROXYPHENYLGLYCINE; MICROBIAL TRANSFORMATION; L-TRYPTOPHAN; MECHANISM;

D O I：

10.1016/0141-0229(95)00044-5

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

A carbamoylase enzyme was purified from a cell-free extract of Agrobacterium sp. with an overall yield of 81%. It was judged to be homogenous on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, with a subunit molecular weight of 38,000 daltons. Further studies on the native enzyme suggested that the active enzyme was present as a dimer, with a pl of 5.5. It was able to cleave a variety of N-carbamoyl substrates, but was strictly D(-) specific. It was found to have a K-m of 0.82 mM and a V-max of 31 U mg(-1) for D(-) N-carbamoyl hydroxyphenylglycine in the presence of 10 mM dithiothreitol. It showed no metal ion requirements but was inhibited by iodoacetic acid and iodoacetamide, both thiol reagents. The N-terminal amino acid sequence of the enzyme was elucidated. (C) 1996 by Elsevier Science Inc.

引用

页码：562 / 571

页数：10

共 31 条

[21] BIOTECHNOLOGICAL PRODUCTION OF UNNATURAL L-AMINO-ACIDS FROM D,L-5-MONOSUBSTITUTED HYDANTOINS .2. L-ALPHA-NAPHTHYLALANINE AND L-BETA-NAPHTHYLALANINE [J].