解脂耶氏酵母中N-乙基马来酰亚胺还原酶的纯化与鉴定

Purification and characterization of N-ethylmaleimide reducing enzyme from Candida lipolytica.

作者信息

Mizugaki M, Nakazawa M, Yamamoto H, Yamanaka H

机构信息

Department of Pharmaceutical Sciences, Tohoku University Hospital, Miyagi.

出版信息

J Biochem. 1989 May;105(5):782-4. doi: 10.1093/oxfordjournals.jbchem.a122744.

DOI:10.1093/oxfordjournals.jbchem.a122744

PMID:2753874

Abstract

N-Ethylmaleimide (NEM) reducing enzyme was purified to homogeneity from cell-free extracts of Candida lipolytica by chromatography techniques. The molecular weight of the native enzyme was estimated to be about 43,000 by gel filtration using Superose 12 and to be 47,000 by SDS-PAGE. This enzyme can use both NADPH and NADH as an electron donor, and catalyzes the reduction of the carbon-carbon double bond of five membered ring compounds which have two conjugated carbonyl groups on both sides of a double bond.

摘要

通过色谱技术从解脂假丝酵母的无细胞提取物中纯化出N-乙基马来酰亚胺（NEM）还原酶，使其达到同质状态。使用Superose 12通过凝胶过滤法估计天然酶的分子量约为43,000，通过SDS-PAGE法估计为47,000。该酶可以使用NADPH和NADH作为电子供体，并催化双键两侧带有两个共轭羰基的五元环化合物的碳-碳双键的还原反应。