东曹HW - 65C：硫酸铵作为一种用于酶纯化的新型柱色谱吸附剂。

Toyopearl HW-65C: ammonium sulfate as a new column chromatographic adsorbent for enzyme purification.

作者信息

Sakihama N, Ohmori H, Sugimoto N, Yamasaki Y, Oshino R, Shin M

出版信息

J Biochem. 1983 Jan;93(1):129-34. doi: 10.1093/oxfordjournals.jbchem.a134146.

DOI:10.1093/oxfordjournals.jbchem.a134146

Abstract

We found that Toyopearl HW-65C gel matrix adsorbed ferredoxin and ferredoxin-NADP+ reductase in the presence of concentrated ammonium sulfate. Ferredoxin was strongly adsorbed on the gel in 80% saturated ammonium sulfate, and ferredoxin-NADP+ reductase was adsorbed in 40% saturated ammonium sulfate. The phenomenon was utilized for purification of ferredoxin and the reductase on a Toyopearl HW-65C: ammonium sulfate column. The technique greatly simplified the early stage of purification of ferredoxin and the reductase. The improved purification methods further involved column treatments with DEAE-Toyopearl 650M and Matrex Red A. The effectiveness of the columns is reported. Since a number of other proteins such as cytochrome c, myoglobin, chymotrypsinogen A, ovalbumin, and glucose oxidase were also adsorbed well in an appropriately concentrated ammonium sulfate solution, the method may be of general use in enzyme purification.

摘要

我们发现，在浓硫酸铵存在的情况下，Toyopearl HW - 65C凝胶基质可吸附铁氧化还原蛋白和铁氧化还原蛋白 - NADP⁺还原酶。铁氧化还原蛋白在80%饱和度的硫酸铵中被强烈吸附在凝胶上，而铁氧化还原蛋白 - NADP⁺还原酶在40%饱和度的硫酸铵中被吸附。该现象被用于在Toyopearl HW - 65C：硫酸铵柱上纯化铁氧化还原蛋白和还原酶。该技术极大地简化了铁氧化还原蛋白和还原酶纯化的早期阶段。改进后的纯化方法还包括用DEAE - Toyopearl 650M和Matrex Red A进行柱处理。报告了这些柱的有效性。由于许多其他蛋白质，如细胞色素c、肌红蛋白、胰凝乳蛋白酶原A、卵清蛋白和葡萄糖氧化酶在适当浓度的硫酸铵溶液中也能被很好地吸附，该方法可能在酶纯化中具有普遍用途。