96微孔板固定鸡肝酯酶研究  

Study on the Immobilization of Chicken Liver Esterase on 96 Microplates

作　　者：Le Tuan Tu 王明珠[1] 徐斐[1] 曹慧[1] 

机构地区：[1]上海理工大学食品质量与安全研究所,上海200093

出　　处：《安徽农业科学》2011年第24期14534-14536,共3页Journal of Anhui Agricultural Sciences

基　　金：上海市科委重点项目(10391901500);上海市科技兴农重点攻关项目(2006第7-4号)

摘　　要：[目的]研究并优化鸡肝酯酶固定化的条件,提高其稳定性,便于在流动注射式生物传感器中使用。[方法]以聚氯乙烯和聚苯乙烯材料的96微孔板为载体,采用物理吸附法固定鸡肝酯酶。通过单因素试验考察时间、温度、酶浓度对固定化酶活力及对甲胺磷农药响应值的影响。[结果]聚苯乙烯材料固定的固定化酶对农药抑制有较高响应值,在最佳固定化时间5 h、温度30℃、酶浓度为30 U/m l的条件下得到的固定化酶吸光值为0.87,1 mg/kg甲胺磷农药对其抑制率为28.39%。[结论]该聚苯乙烯材料对鸡肝酯酶具有较好的吸附能力,且固定方法简单,酶分子空间构型稳定,是一种优良的载体。[Objective] The immobilization condition of chicken liver esterase was researched and optimized in order to improve its stability as well as its convenient use in flow injection biosensors.[Method] 96 Microplates of both polyvinyl chloride and polystyrene were used as vector and the chicken liver esterase was immobilized with absorption method.The effect of time,temperature and esterase concentration on the activity of immobilized enzyme and the sensitivity to methamidophos inhibition were studied.[Result] There was high responsible value of the enzyme immobilized by polystyrene to the sensitivity of agro-chemical and under the best immobilization condition that the time was 5 hours,the temperature was 30℃ and the concentration of esterase was 30 U/ml,respectively,the OD absorbance of immobilized enzyme was 0.87 and the inhibition percentage of methamidophos pesticideat 1mg/L was 28.39%.[Conclusion] The polystyrene,a good vector,had better the absorption ability in chicken liver esterase and the method was simple way,which spatial structure of the enzyme molecular was stable.

关 键 词：固定化酶 鸡肝酯酶 吸附 96微孔板 

分 类 号：S188.4[农业科学—农业基础科学]