利用甘露聚糖凝胶亲和层析法改进米曲霉1,2-α-甘露糖苷酶的纯化

Improved purification of Aspergillus oryzae 1,2-alpha-mannosidase by using mannan gel affinity chromatography.

作者信息

Tanimoto K, Nishimoto T, Saitoh F, Yamaguchi H

出版信息

J Biochem. 1986 Feb;99(2):601-4. doi: 10.1093/oxfordjournals.jbchem.a135517.

DOI:10.1093/oxfordjournals.jbchem.a135517

Abstract

In order to facilitate the purification of 1,2-alpha-mannosidase from an enzyme product of Aspergillus oryzae, we have devised a rapid and simple procedure. A partially purified enzyme preparation obtained from the A. oryzae enzyme product, by means of ammonium sulfate fractionation followed by CM-Sephadex C-50 chromatography, was subjected to affinity chromatography with baker's yeast mannan gel as an adsorbent. 1,2-alpha-Mannosidase was retarded and well separated from the major protein peak on the affinity column. After a second affinity chromatography under the same conditions, 1,2-alpha-mannosidase was finally purified 7,500-fold with a 22.9% yield. The enzyme preparation thus obtained was quite suitable for the structural analysis of glycoconjugates.

摘要

为了便于从米曲霉的酶产物中纯化1,2-α-甘露糖苷酶，我们设计了一种快速简便的方法。通过硫酸铵分级分离，然后进行CM-葡聚糖凝胶C-50柱色谱，从米曲霉酶产物中获得部分纯化的酶制剂，以面包酵母甘露聚糖凝胶为吸附剂进行亲和色谱。1,2-α-甘露糖苷酶在亲和柱上被阻滞，并与主要蛋白峰良好分离。在相同条件下进行第二次亲和色谱后，1,2-α-甘露糖苷酶最终纯化了7500倍，产率为22.9%。由此获得的酶制剂非常适合用于糖缀合物的结构分析。