植物酯酶分离纯化及酶促反应条件优化的研究  

Study on Isolation and Purification of Plant Esterase and Enzymatic Reaction Conditions Optimization

作　　者：陈士恩[1] 申晓蓉[1] 罗振兴[1] 霍生东[1] 阿依木古丽[1] 刘丽霞[1] 

机构地区：[1]西北民族大学生命科学与工程学院,甘肃兰州730030

出　　处：《西北民族大学学报（自然科学版）》2012年第1期41-46,72,共7页Journal of Northwest Minzu University(Natural Science)

基　　金：校地合作项目<早胜肉牛优质高效综合配套技术体系应用研究与产业化>(XBMV-2008-BC-24)

摘　　要：目的:建立一种快速检测牛肉中有机磷农药残留的方法.方法:从新鲜小麦粉、黄豆粉和玉米粉中分离植物酯酶,真空冷冻浓缩,羧甲基纤维素层析,真空冷冻干燥提取植物酯酶,并通过试验测得其酶促反应最适温度36.8℃,最适pH 7.7,酶促反应时间3 min,敌敌畏对其抑制时间8 min,显色剂2,6-二氯靛酚的最大吸收波长606 nm.结论:在相同条件下,提纯后的植物酯酶检测模拟有机磷农药残留阳性牛肉样品抑制率为99.5%,市售丁酰胆碱酯酶测得的抑制率为99.8%%,两者检测结果高度一致.Objectives： It was to establish a rapid method of detection organophosphorus pesticide residues in beef. Methods Plant esterase was separated from fresh in wheat flour, vacuum freezing enrichment, carboxymethyl cellulose chromatography. The wheat esterase was extracted using vacuum freeze - drying. Results The enzymatic reactions were the optimum temperature for 36.8 ＊C, the optimal pH for 7.7, enzymatic reaction time for 3min, panaplate on its inhibition time for 8mln, colour- display reagent utilization leuco - 2, 6 - dichlorophenolindophenol the maximum absorption wavelength for 606 nm. Conclusion Iin the same condition, the purification of wheat esterase was after testing positive for organophosphorus pesticide residues simulated sample inhibition rate for beef, 99.5 %, Market sales butyl acylating cholinesterase measurement of the inhibition rate was 99.8 %, both test results height consistent.

关 键 词：植物酯酶 分离纯化 酶促反应 最适反应条件 

分 类 号：S851.4[农业科学—预防兽医学]