摘要
首次采用以改性纤维素为基质、亚氨二乙酸（ＩＤＡ）为取代配基的铜离子螯合膜色谱法对牛肝过氧化氢酶（ＢＬＣ）的分离纯化进行了研究。缓冲液的ｐＨ值对ＢＬＣ与螯合配基的结合影响显著。在选定的色谱条件下，ＢＬＣ粗酶液经ＩＤＡ型Ｃｕ２＋－螯合膜色谱柱一步纯化，比活性平均提高４．７倍，回收率为６７．７％。金属螯合膜色谱柱可用含０．２ｍｏｌ／Ｌ的咪唑或５０ｍｍｏｌ／ＬＥＤＴＡ－１ｍｏｌ／ＬＮａＣｌ的缓冲液再生，反复使用，后者比前者对柱子的再生效果更好。
Cu 2+ 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 Cu 2+ 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.
出处
《色谱》
CAS
CSCD
北大核心
1997年第4期292-295,共4页
Chinese Journal of Chromatography
基金
国家自然科学基金
中科院大连化学物理研究所科研基金
关键词
亲和色谱法
金属螯合膜
过氧化氢酶
纯化
牛肝
affinity chromatography, metal ion chelating membrane, catalase, purification
