泛解硫胺素酶活性的连续分光光度测定法。

Continuous spectrophotometric assay of pantetheinase activity.

作者信息

Duprè S, Chiaraluce R, Nardini M, Cannella C, Ricci G, Cavallini D

出版信息

Anal Biochem. 1984 Oct;142(1):175-81. doi: 10.1016/0003-2697(84)90534-7.

Abstract

A continuous spectrophotometric assay for pantetheinase determination using S-pantetheine-3-pyruvate as substrate is described. The enzymatic hydrolysis of this new substrate leads to the formation of S-cysteamine-3-pyruvate, which cyclizes in a non-rate-limiting step to give 2H-1,4-thiazin-5,6-dihydro-3-carboxylic acid (aminoethylcysteine ketimine), a compound exhibiting a strong absorption at 296 nm. The assay is optimized with respect to pH, buffer, and substrate concentration. Prereduction of the enzyme and some properties of the reaction are also studied. The assay is simple, rapid, very sensitive, and specific.

摘要

描述了一种使用S-泛酰巯基乙胺-3-丙酮酸作为底物的连续分光光度法测定泛肽酶。这种新底物的酶促水解导致形成S-半胱胺-3-丙酮酸，其在一个非限速步骤中环化生成2H-1,4-噻嗪-5,6-二氢-3-羧酸（氨基乙基半胱氨酸酮亚胺），该化合物在296nm处有强烈吸收。该测定法在pH、缓冲液和底物浓度方面进行了优化。还研究了酶的预还原及反应的一些性质。该测定法简单、快速、非常灵敏且具有特异性。