APPLICATION OF HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY TO THE STUDY OF THE BIOLOGICAL TRANSFORMATION OF ADIPONITRILE

被引:2
作者
MOREAU, JL [1 ]
AZZA, S [1 ]
BIGEY, F [1 ]
ARNAUD, A [1 ]
GALZY, P [1 ]
机构
[1] ECOLE NATL SUPER AGRON MONTIPELLIER,CHAIR MICROBIOL IND & GENET MICOORGANISMES,F-34060 MONTPELLIER,FRANCE
来源
JOURNAL OF CHROMATOGRAPHY B-BIOMEDICAL APPLICATIONS | 1994年 / 656卷 / 01期
关键词
D O I
10.1016/0378-4347(94)00091-3
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A procedure for the assay of nitrile hydratase and amidase activity by high-performance liquid chromatography is described. The method can be used to assay the intermediate compounds resulting from the hydrolysis of adiponitrile into adipic acid, and to determine the kinetics of the hydrolysis of these compounds using whole cells and enzyme extracts. The precision of the method makes it suitable for the determination of the enzyme parameters: K-m and V-m (nitrile hydratase and amidase). Using cyanovaleramide as substrate, K-m and V-m were respectively 370 mM and 2060 U/mg for nitrile hydratase and 6.6 mM and 33 U/mg for amidase.
引用
收藏
页码:197 / 202
页数:6
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