猪脑乙酰胆碱酯酶的分离纯化和性质研究  

Purification and Characterization of Acetylcholinesterase from Porcine Brain

作　　者：王洁[1] 李星[1] 王红扬[1] 唐云明[1] 

机构地区：[1]西南大学生命科学学院重庆市甘薯工程研究中心三峡库区生态环境教育部重点实验室,重庆400715

出　　处：《食品科学》2015年第3期137-141,共5页Food Science

基　　金：重庆市科委重点攻关项目(CSTC2011AB1027)

摘　　要：通过离心、硫酸铵分级沉淀、DEAE-Sepharose和Superdex-200层析等步骤,从猪脑中获得电泳纯乙酰胆碱酯酶,该酶比活力和纯化倍数分别为2.05 U/mg和26.97,酶活回收率为11.95%;该酶分子质量为257.30 kD,亚基为66.94 kD;以碘化硫代乙酰胆碱为底物时,酶的最适反应温度为37 ℃,最适pH值为7.4,且在40 ℃以下,pH 6.0～8.0有较好的稳定性;最适底物浓度为4.0 mmol/L,Km为0.94 mmol/L。Ba2+和Zn2+对该酶有强烈的抑制作用,而低浓度Mg2+对该酶有激活作用。Acetylcholinesterase (AChE) was purified from porcine brain by consecutive steps including centrifugation, ammonium sulfate fractionation, DEAE-Sepharose chromatography and Superdex-200 gel filtration chromatography. Thepurified AChE exhibited a specific activity of 2.05 U/mg, with 26.97-fold purification and an activity yield of 11.95%. The relative molecular weight of AChE was 257.30 kD, and the subunit molecular weight was 66.94 kD. The optimal pH and temperature for the enzyme were 7.4 and 37 ℃, respectively. The enzyme was stable below 40 ℃ and in the pH range of 6.0–8.0. At an optimal substrate concentration of 4.0 mmol/L, the apparent Km was 0.94 mmol/L. The activity of AChE was inhibited by Ba2+ or Zn2+ , but enhanced by Mg2+.

关 键 词：乙酰胆碱酯酶 猪脑 分离纯化 性质 

分 类 号：TS251.9[轻工技术与工程—农产品加工及贮藏工程]