摘要
传统对血清中胆碱酯酶的分离纯化步骤繁琐且纯化倍数低,本实验采用硫酸铵分级沉淀、透析,再经DEAE-52离子交换层析和SephadexG-100凝胶过滤层析的方法,对马血清中胆碱酯酶进行分离纯化,得到胆碱酯酶纯品,纯化倍数达101.3倍。经SDS-PAGE不连续凝胶电泳显示为单一条带,并测定其相对分子量为72000Da。胆碱酯酶性质研究的实验结果表明,该酶最适温度为35℃,最适pH为7.4,对底物氯化乙酰胆碱的Km值为0.192mmol/L,最适底物浓度为4mmol/L,高浓度底物对酶有抑制作用,该酶对热不稳定。
Traditional separation and purification of cholinesterase from serum was complicated insufficient. In this experiment cholinesterase from horse serum was purified by ammonium sulfate precipitation, ion-exchange chromatography on DEAE-52 and gel filtration on Sephadex G-100. The enzyme was purified by up to 101.3 times. Purified ChE showed only one band on SDS- PAGE with a molecular weight of about 72,000 Dalton. Studies on the properties of the Cholinesterase showed that the optimum temperature and pH value were 35℃ and 7.4, respectively. The Km value of acetylcholine chloride was 0.192mmol/L. The optimum concentration of substrate was 4mmol/L, and the enzyme could be inhibited by high level of acetylcholine chloride. The enzyme was sensitive to high temperature.
出处
《食品工业科技》
CAS
CSCD
北大核心
2006年第9期91-93,共3页
Science and Technology of Food Industry
关键词
胆碱酯酶
马血清
纯化
cholinesterase
-horse serum
purification
