通过在蓝色葡聚糖-琼脂糖4B上进行亲和层析从兔肝脏中高效纯化乙二醛酶I。

A high-yield purification of glyoxalase i from rabbit liver by affinity chromatography on Blue Dextran-Sepharose 4B.

作者信息

Elango N, Srinivasan A, Rao A R

出版信息

J Biochem Biophys Methods. 1981 Mar;4(3-4):233-40. doi: 10.1016/0165-022x(81)90061-0.

Abstract

An improved method is described for the purification of glyoxalase I from rabbit liver. The method involves homogenization, ammonium sulfate precipitation followed by choloroform/ethanol precipitation, affinity chromatography on Blue Dextran-Sepharose 4B and chromatography on Sephadex G-100. The enzyme is specifically eluted from the affinity column by S-hexylglutathione, a competitive inhibitor of the enzyme. This procedure offers a convenient method for obtaining electrophoretically pure glyoxalase I in high yields (60--70%).

摘要

描述了一种从兔肝中纯化乙二醛酶I的改进方法。该方法包括匀浆、硫酸铵沉淀，随后进行氯仿/乙醇沉淀、在蓝色葡聚糖-琼脂糖4B上进行亲和层析以及在葡聚糖凝胶G-100上进行层析。该酶通过S-己基谷胱甘肽（一种酶的竞争性抑制剂）从亲和柱上特异性洗脱。此方法为以高产率（60% - 70%）获得电泳纯的乙二醛酶I提供了一种便捷的方法。