豌豆酯酶的纯化和酯酶活性表征及其对农药的敏感性  

Purification,characterization and pesticides-sensitivity of plant-esterase from Pisum sativum Linn.

作　　者：黄嫒 马志强 王玮 宋燕飞 李荣 陈强 陈祥贵 杨潇 HUANG Ai;MA Zhiqiang;WANG Wei;SONG Yanfei;LI Rong;CHEN Qiang;CHEN Xianggui;YANG Xiao(School of Food and Biological Engineering,Xihua University,Chengdu 610039,China)

机构地区：[1]西华大学食品与生物工程学院

出　　处：《食品与发酵工业》2019年第11期78-84,共7页Food and Fermentation Industries

基　　金：国家自然科学基金面上项目(31271872)

摘　　要：由于食品和环境中的农药残留对人体健康构成威胁,使得发展农残快速检测方法显得十分重要,从植物中发现新的酶源有助于降低酶抑制法快速检测农药的成本。实验从豌豆中分离纯化豌豆酯酶,并进行了酶类鉴别、特性表征和农药敏感性研究。纯化后的豌豆酯酶分子质量约为26.0kDa,具有较高的酯酶活性。通过底物和抑制剂特异性试验,将豌豆酯酶分类为羧酸酯酶。豌豆酯酶在质量浓度为0.45μg/mL,缓冲液pH6.5和反应温度40℃时,表现出最好的催化活性。豌豆酯酶可以被有机磷和氨基甲酸酯类农药抑制。豌豆酯酶属于B型酯酶,可用于酶抑制法检测有机磷和氨基甲酸酯类农药残留,且经过纯化的豌豆酯酶满足大多数农药最大残留限值(MRL)检测的要求。Pesticide residues in foods and environment pose threats to human health,therefore,discovering new enzyme sources from plants can help reduce the cost of rapid pesticide detection by enzymatic inhibition method. Plant esterases from peas were purified,classified and characterized. Their pesticide sensitivities were also detected. The results showed that the purified pea esterase was about 26.0 kDa with high esterase activity,and it was classified as a carboxylesterase. Moreover,it demonstrated the best catalytic activity at 40 ℃ and pH 6.5 at 0.45 μg/mL. Furthermore,it could be inhibited by organophosphate and carbamate pesticides. In conclusion,pea esterase is a type B esterase and can be used to detect organophosphorus and carbamate pesticide residues. Besides,the purified pea esterase also meets the requirements of most pesticide MRL tests.

关 键 词：植物酯酶 豌豆 酶活表征 有机磷和氨基甲酸酯类农药 酶抑制法 

分 类 号：S481.8[农业科学—农药学]