用于农药残留检测的大豆酯酶的纯化分离

PURIFICATION AND SEPARATION OF SOYBEAN ESTERASE FOR PESTICIDE RESIDUE DETECTION

以大豆种子为原料探索农药残留检测用酯酶的新酶原。采用差素离心、硫酸铵盐析、DE-AE-32离子交换层析和SephadexG-100凝胶过滤层析等方法对大豆酯酶进行了纯化分离,并研究了纯化酶对有机磷和氨基甲酸酯类农药的敏感性。最终得到3种电泳纯的大豆酯酶,记为E1,E2和E3。但3种大豆酯酶对有机磷和氨基甲酸酯类农药的敏感性不同,E1对农药没有敏感性,E2的敏感性比E3好。因此,大豆酯酶E2可作为农药残留检测的新酶原。

To search a new esterase for pesticide residue detection from soybean seeds,the soybean seed powder was dealt with crude extraction,differential centrifugation,ammonium sulfate salting-out to purify soybean esterase. The purified enzyme was then separated by DEAE-32 ion exchange column chromatography and Sephadex G-100 gel filtration column chromatography. Then,the inhibition to Organophosphate and Carbamate pesticides of the esterase was observed. Three soybean esterase of eleetrophoresis purity were obtained, marked with E1, E2 and E3. Rusults indicated that E1 had no sensitivity （inhibition） to Organophosphate and Carbamate pesticide while E2 was better sensitive than E3. Hence,a new esterase for pesticide residue detection from soybean seeds and a better purification method were obtained.