Studies on the Purification and Characterization of Soybean Esterase,and Its Sensitivity to Organophosphate and Carbamate Pesticides  

作　　者：LI Jian-ke ZHOU Yan-li WEN Yan-xia WANG Jian-hua HU Qiu-hui 

机构地区：[1]College of Food Engineering and Nutritional Science, Shaanxi Normal University, Xi＇an 710062, P.R.China [2]College of Veterinary Medicine, Northwest A＆F Univerisity, Yangling 712100, P.R. China [3]College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, P.R.China

出　　处：《Agricultural Sciences in China》2009年第4期455-463,共9页中国农业科学（英文版）

基　　金：the financial supports from the Ministry of Science and Technology of China(2006BAK02A24);the Ministry of Agriculture of China(2007-2011);Xi'an Bureau of Science and Technology,China (GG05112)

摘　　要：Soybean esterase, a cholinesterase-like enzyme, was purified by differential centrifugation firstly, then, ammonium sulfate precipitation, dialysis, and finally, DEAE-cellulose-32 ion-exchange chromatography after extracting it from soybean seeds with phosphate buffer （0.3 mol L^-1, pH 7.0）. The extract recovery rate of the purified enzyme was 8.18% and purification fold was 91.58. The soybean esterase appeared as two bands on the denaturing SDS-PAGE with molecular weights of 24 and 37.2 kDa, respectively, which proved that it is a dimer protein consisting of two subunits. The result of nondenaturing PAGE revealed that the soybean esterase is a single band with cholinesterase-like activity using α- naphthyl acetate as the substrate and fast blue B salt as coloring agent. The esterase showed very high sensitivity to 18 kinds of organophosphate pesticides and 6 kinds of carbamate pesticides with the lowest detective limits of 0.03125- 0.0625 and 0.03125-0.25 mg kg^-1, respectively, and can meet the demands of MRL specified by the most countries.Soybean esterase, a cholinesterase-like enzyme, was purified by differential centrifugation firstly, then, ammonium sulfate precipitation, dialysis, and finally, DEAE-cellulose-32 ion-exchange chromatography after extracting it from soybean seeds with phosphate buffer （0.3 mol L^-1, pH 7.0）. The extract recovery rate of the purified enzyme was 8.18% and purification fold was 91.58. The soybean esterase appeared as two bands on the denaturing SDS-PAGE with molecular weights of 24 and 37.2 kDa, respectively, which proved that it is a dimer protein consisting of two subunits. The result of nondenaturing PAGE revealed that the soybean esterase is a single band with cholinesterase-like activity using α- naphthyl acetate as the substrate and fast blue B salt as coloring agent. The esterase showed very high sensitivity to 18 kinds of organophosphate pesticides and 6 kinds of carbamate pesticides with the lowest detective limits of 0.03125- 0.0625 and 0.03125-0.25 mg kg^-1, respectively, and can meet the demands of MRL specified by the most countries.

关 键 词：SOYBEAN phytoesterase PESTICIDES ORGANOPHOSPHATE CARBAMATE 

分 类 号：S436.3[农业科学—农业昆虫与害虫防治] Q556.1[农业科学—植物保护]