中华蜜蜂头部乙酰胆碱酯酶最佳反应体系的建立  

Establishment of optimization of assaying activity of head acetylcholinesterase from Apis cerana cerana

作　　者：王宇豪[1,2] 赵冬香[1] 王玉洁[1] 刘俊峰[1] 高景林[1] 刘桂琼[2] 

机构地区：[1]中国热带农业科学院环境与植物保护研究所,海口571101 [2]华中农业大学动物科技学院动物医学院,武汉430070

出　　处：《中国蜂业》2015年第11期20-24,共5页Apiculture of China

基　　金：国家蜂产业体系儋州综合实验站建设项目(No.CARSSYZ-14);海南省应用技术研发与示范推广专项(No.ZDXM2014050);公益性行业(蜜蜂)科研专项子课题(No.201203080-8)

摘　　要：乙酰胆碱酯酶是昆虫体内重要的生化标记物,利用其活性的变化可以监测农药对昆虫的亚致死效应。乙酰胆碱酯酶活性测定的条件会因为来源于不同的物种甚至同一物种不同部位而改变。本文采用正交试验排列的25个组合试验探究了酶浓度、底物浓度、pH、反应温度及反应时间5个因素对测定中华蜜蜂头部乙酰胆碱酯酶比活力的影响,并从中选择出最佳测定组合。试验结果表明,5个因素对中华蜜蜂头部乙酰胆碱酯酶比活力影响的大小顺序为酶浓度>反应温度>pH>反应时间>底物浓度,各因素最佳组合为酶浓度0.25g/L、底物浓度0.8mmol/L、pH值7.5、反应温度40℃及反应时间5min。本文优化了中华蜜蜂头部乙酰胆碱酯酶测定条件,为今后评估有机磷类、氨基甲酸类和新烟碱类杀虫剂提供条件。Acetylcholinesterase is an important biochemical marker in insects, and its changes are usually used to monitor pesticide sublethal effects of insects. The conditions of acetylcholinesterase activity will change with different species or different parts of the same species In this paper, we identified the enzyme Concentration, substrate concentration, pH, reaction temperature and reaction time by 25 combinations of orthogonal experiment. The five factors influence the specific activity of the head acetylcholinesterase of the Apis cerana cerana. The results showed that the order of the activity of acetylcholinesterase followed as concentration of protein 〉 temperature 〉 pH 〉 time 〉 concentration of substrate. And the optimal conditions were listed, such as enzyme concentration 0.25g protein /L, substrate concentration 0.8 mmol /L, pH 7.5, reaction temperature 40℃ and reaction time 5 min. Above all, we optimized the conditions of A. cerana cerana head acetylcholinesterase, which will be helpful to the assessment of organophosphorus, carbamate and neonicotinoid insecticides.

关 键 词：中华蜜蜂 乙酰胆碱酯酶 正交试验 最佳测定条件 

分 类 号：S891[农业科学—特种经济动物饲养]