酿酒酵母中sn-甘油-3-磷酸脱氢酶的纯化改进及其一些分子和动力学特性

Improved purification and some molecular and kinetic properties of sn-glycerol-3-phosphate dehydrogenase from Saccharomyces cerevisiae.

作者信息

Chen S M, Trumbore M W, Osinchak J E, Merkel J R

机构信息

Department of Chemistry, Lehigh University, Bethlehem, PA 18015.

出版信息

Prep Biochem. 1987;17(4):435-46. doi: 10.1080/00327488708062506.

DOI:10.1080/00327488708062506

PMID:3325967

Abstract

The purification procedure for isolating sn-glycerol-3-phosphate dehydrogenase (EC 1.1.1.8) from Saccharomyces cerevisiae was improved by the introduction of an ion-exchange step. Enzyme yields were doubled and the specific activity was increased as compared to the original procedure. A new value of 42,000 was obtained for the molecular weight by several denaturing methods. By native gel chromatography the molecular weight appears to be 31,000 as reported earlier. Michaelis constants were found to be 0.37 mM with dihydroxyacetone phosphate as the variable substrate and 0.018 mM for NADH as the variable substrate.

摘要

通过引入离子交换步骤，改进了从酿酒酵母中分离sn-甘油-3-磷酸脱氢酶（EC 1.1.1.8）的纯化程序。与原始程序相比，酶产量提高了一倍，比活性也有所增加。通过几种变性方法测得分子量的新值为42,000。通过天然凝胶色谱法，分子量似乎如先前报道的为31,000。发现以磷酸二羟丙酮作为可变底物时米氏常数为0.37 mM，以NADH作为可变底物时米氏常数为0.018 mM。