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Position: Home > Articles > Purification and Characterization of Acetylcholinesterase from Porcine Brain FOOD SCIENCE 2015,36 (3) 137-141

猪脑乙酰胆碱酯酶的分离纯化和性质研究

作  者:
王洁;李星;王红扬;唐云明
单  位:
西南大学生命科学学院;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
关键词:
乙酰胆碱酯酶;猪脑;分离纯化;性质
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。
译  名:
Purification and Characterization of Acetylcholinesterase from Porcine Brain
作  者:
WANG Jie;LI Xing;WANG Hongyang;TANG Yunming;Key Laboratory of Eco-environments in Three Gorges Reservoir Region, Ministry of Education, Chongqing Sweetpotato Engineering Research Center, College of Life Science, Southwest University;
关键词:
acetylcholinesterase;;porcine brain;;isolation and purification;;characterization
摘  要:
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+.

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