高效液相色谱法在己二腈生物转化研究中的应用。

Application of high-performance liquid chromatography to the study of the biological transformation of adiponitrile.

作者信息

Moreau J L, Azza S, Bigey F, Arnaud A, Galzy P

机构信息

Chaire de Microbiologie Industrielle et de Génétique des Microorganismes, Ecole Nationale Supériure Agronomique de Montpellier, France.

出版信息

J Chromatogr B Biomed Appl. 1994 Jun 3;656(1):197-202. doi: 10.1016/0378-4347(94)00091-3.

DOI:10.1016/0378-4347(94)00091-3

PMID:7952029

Abstract

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: Km and Vm (nitrile hydratase and amidase). Using cyanovaleramide as substrate, Km and Vm were respectively 370 mM and 2060 U/mg for nitrile hydratase and 6.6 mM and 33 U/mg for amidase.

摘要

描述了一种通过高效液相色谱法测定腈水合酶和酰胺酶活性的方法。该方法可用于分析己二腈水解生成己二酸过程中产生的中间化合物，并使用全细胞和酶提取物测定这些化合物的水解动力学。该方法的精密度使其适用于酶参数的测定：Km和Vm（腈水合酶和酰胺酶）。以氰基戊酰胺为底物时，腈水合酶的Km和Vm分别为370 mM和2060 U/mg，酰胺酶的Km和Vm分别为6.6 mM和33 U/mg。