使用新型封闭底物（3-氧代丁叉基-β-2-氯-4-硝基苯基麦芽五糖苷）测定α-淀粉酶

Determination of alpha-amylase using a new blocked substrate (3-ketobutylidene beta-2-chloro-4-nitrophenyl-maltopentaoside).

作者信息

Teshima S, Hayashi Y, Emi S, Ishimaru K

机构信息

Tsuruga Enzyme Plant, Toyobo Co. Ltd., Fukui and Japan.

出版信息

Clin Chim Acta. 1991 May 31;199(1):23-31. doi: 10.1016/0009-8981(91)90005-w.

DOI:10.1016/0009-8981(91)90005-w

PMID:1934499

Abstract

A new substrate, 3-ketobutylidene beta-2-chloro-4-nitrophenylmaltopentaoside (3KB-CNPG5), was used for the determination of alpha-amylase (EC 3.2.1.1) in serum and urine. Under this alpha-amylase assay condition, 3KB-CNPG5 is resistant to glucoamylase and alpha-glucosidase, which are auxiliary enzymes, because the 4- and 6-positions of the non-reducing-end glucose residue are modified by the 3-ketobutylidene group. The assay using 3KB-CNPG5 for alpha-amylase activity is a highly sensitive method that uses 2-chloro-4-nitrophenol (CNP) as an aglycone, and is a stable method for determination of alpha-amylase activity in biological fluids.

摘要

一种新的底物，3-酮丁叉基-β-2-氯-4-硝基苯基麦芽五糖苷（3KB-CNPG5），被用于测定血清和尿液中的α-淀粉酶（EC 3.2.1.1）。在这种α-淀粉酶测定条件下，3KB-CNPG5对作为辅助酶的葡糖淀粉酶和α-葡糖苷酶具有抗性，这是因为非还原端葡萄糖残基的4位和6位被3-酮丁叉基修饰。使用3KB-CNPG5测定α-淀粉酶活性是一种高度灵敏的方法，该方法以2-氯-4-硝基苯酚（CNP）作为糖苷配基，也是一种用于测定生物体液中α-淀粉酶活性的稳定方法。