摘要
以Ellman法测定锦龟头部酯酶酶活,以吲哚乙酸酯为底物测定树鼩肝部酯酶酶活,并用正交试验确定最佳反应体系条件,比较几种特异性酶抑制剂的抑制效果;采用酶抑制法考察锦龟酯酶和树鼩酯酶对有机磷农药和氨基甲酸酯类农药的敏感性。结果表明:锦龟酯酶酶活测定方法的最佳条件为孵育温度35℃、孵育时间15min、缓冲液pH 7.5;树鼩酶酶活测定的最佳条件为孵育温度25℃、孵育时间15min、缓冲液pH 8.0;锦龟酯酶对盐酸多奈哌齐敏感性最高;树鼩脂酶对双(4-硝基苯基)磷酸酯敏感性最高;锦龟酯酶和树鼩酯酶对敌百虫、辛硫磷、敌敌畏、速灭磷、磷铵、速灭威、克百威、灭多威、涕灭威、甲萘威的检出限均低于国标所规定的农药最大残留量限量。锦龟酯酶与树鼩酯酶粗提物具有良好的农药敏感性,可作为新型酶源用于酶抑制法检测有机磷和氨基甲酸酯类农药残留。
The esterase activity of Chrysemyswas determined by Ellman method,and the indole acetate was used as substrate to determine the esterase activity of Tupaiabelangeri.The optimum reaction conditions were determined by orthogonal array design.and the inhibition effects of several specific enzyme inhibitors were compared;The enzyme inhibition method was used to investigated the sensitivity of esterase form Chrysemyse and Tupaiabelangeri to organophosphorus and carbamate pesticides.The optimum conditions of esterase activity determination of Chrysemys were:incubation temperature 35 ℃,incubation time15 min,buffer pH 7.5;The optimum conditions of esterase activity determination of Tupaiabelangeri were:incubation temperature 25 ℃,incubation time 15 min,buffer pH 8.0.The most sensitive to Chrysemysesterase was donepezil hydrochloride,the most sensitive to Tupaiabelangeri esterasewas bis(4-nitrophenyl)phosphate.The detection limits of esterase form Chrysemys and Tupaiabelangeri on trichlorfon,phoxim,dichlorvos,mevinphos,phosphamidon,metolcarb,carbofuran,methomyl,aldicarb and carbaryl were all lower than the maximum residue limits stipulated by national standards.The extracts of esterase from Chrysemys and Tupaiabelangeriwere showed high sensitivity to pesticide,which could be used as a new enzyme source in the enzyme inhibition method for rapid detection organophosphorus and carbamate pesticide.
作者
宋燕飞
陈祥贵
向俊丞
王玮
阴鹏飞
杨潇
无
SONG Yan-fei;CHEN Xiang-gui;XIANG Jun-cheng;WANG Wei;YIN Peng-fei;YANG Xiao(School of Food and Biological Engineering,Xihua University,Chengdu,Sichuan 610039,China;Yibin Xihua University Research Institute,Yibin,Sichuan 644004,China)
出处
《食品与机械》
北大核心
2019年第11期58-63,共6页
Food and Machinery
基金
国家自然基金项目（编号：31271872）
四川省科技厅科技计划项目（编号：18MZGC0276）
四川省科技厅重点研发项目计划资助（编号：2018SZ0340)
关键词
锦龟
树鼩
农药
敏感性
Chrysemys
Tupaiabelangeri
pesticide
sensitivity
