用于纯化牛肝过氧化氢酶的固定化IDA型铜离子螯合膜亲和色谱法

Immobilized IDA-type Cu2+-chelating membrane affinity chromatography for purification of bovine liver catalase.

作者信息

Yang L, Jia L, Zou H, Kong L, Zhang Y

机构信息

National Chromatographic R. & A. Center of China, Dalian Institute of Chemical Physics, the Chinese Academy of Sciences, Dalian, 116011.

出版信息

Se Pu. 1997 Jul;15(4):292-5.

PMID:15739458

Abstract

Cu2+-chelating membrane medium based on iminodiacetate-substituted modified short cotton cellulose was examined to purify bovine liver catalase (BLC) by affinity chromatography. The buffer pH notably affect BLC binding to the Cu2+-chelating membrane adsorbent. The results showed that under the chromatographic conditions chosen, 67.7% recovery of BLC attained with an overall 4.7-fold increase in specific activity in a single step. After purification of BLC, the chelating membrane adsorbent can easily be regenerated by imidazole or EDTA buffer. Different results were obtained by the two regeneration methods: the later is more effective than the former in reviving the purification characters of the chelating membrane adsorbent.

摘要

研究了基于亚氨基二乙酸取代的改性短棉纤维素的Cu2+螯合膜介质，通过亲和色谱法纯化牛肝过氧化氢酶（BLC）。缓冲液pH值显著影响BLC与Cu2+螯合膜吸附剂的结合。结果表明，在所选择的色谱条件下，一步法可实现BLC回收率达67.7%，比活总体提高4.7倍。纯化BLC后，螯合膜吸附剂可通过咪唑或EDTA缓冲液轻松再生。两种再生方法得到了不同的结果：后者在恢复螯合膜吸附剂的纯化特性方面比前者更有效。