植物酯酶分离纯化及酶促反应条件优化的研究

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.

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

Objectives:It was to establish a rapid method of detection organophosphorus pesticide residuesin beef.Methods Plant esterase was separated fromfreshin wheat flour,vacuumfreezing enrichment,car-boxymethyl cellulose chromatography.The wheat esterase was extracted using vacuumfreeze-drying.Re-sults The enzymatic reactions were the opti mumtemperaturefor 36.8 ℃,the opti mal pHfor 7.7,enzymat-ic reaction ti me for 3min,panaplate onitsinhibitionti me for 8min,colour-display reagent utilizationleuco-2,6-dichlorophenolindophenol the maxi mumabsorption wavelengthfor 606 nm.ConclusionIinthe samecondition,the purification of wheat esterase was after testing positive for organophosphorus pesticideresidues si mulated sample inhibition rate for beef,99.5 %,Market sales butyl acylating cholinesterase mea-surement of the inhibition rate was 99.8 %,both test results height consistent.