Purification and characterization of the D-hydantoinase from Bacillus circulans

被引：7

作者：

Luksa, V ^{[1
]}

Starkuviene, V ^{[1
]}

Starkuviene, B ^{[1
]}

Dagys, R ^{[1
]}

机构：

[1] INST BIOTECHNOL FERMENTAS,LT-2028 VILNIUS,LITHUANIA

来源：

APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY | 1997年 / 62卷 / 2-3期

关键词：

D-hydantoinase; Bacillus circulans; thermostability; SDS-treatment;

D O I：

10.1007/BF02787998

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A D-hydantoinase (5,6-dihydropyrimidine amidohydrolase) was purified to homogeneity from Bacillus circulans. Purification of two hundred forty-three-fold was achieved with an overall yield of 12%. The relative molecular mass of the native enzyme is 212,000 and that of the subunit is 53,000. This enzyme is an acidic protein with an isoelectric point of 4.55. The enzyme is sensitive to thiol reagent and requires metal ions for its activity. The optimal conditions for the hydantoinase activity are pH 8.0-10.0 and a temperature of 75 degrees C. The enzyme is the most stable in a pH range of 8.5-9.5 and up to 60 degrees C. The enzyme is significantly stable not only at high temperatures but also on treatment with protein denaturant SDS. These remarkable properties are used for the purification procedure.

引用

页码：219 / 231

页数：13

共 19 条

[1]

BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3

[2] THERMOSTABLE DIPEPTIDASE FROM BACILLUS-STEAROTHERMOPHILUS - ITS PURIFICATION, CHARACTERIZATION, AND COMPARISON WITH AMINOACYLASE [J].