摘要
[目的]探索滤纸壳聚糖膜固定乙酰胆碱酯酶(AChE)的最佳条件。[方法]以滤纸壳聚糖膜为载体,戊二醛为交联剂,牛血清白蛋白(BSA)为保护剂,进行AChE的固定。并对固定化酶的理化性质进行研究。[结果]固定AChE的最佳条件为将50μl 150 U/mlAChE液,50μl 5%(V/V)戊二醛溶液,100μl 1%(W/W)BSA,pH 8.0的0.2 mol/L的PBS缓冲液配制成1 ml酶液,滤纸壳聚糖膜浸于该酶液4℃固定8 h。固定化酶的最适反应温度为37℃,最适pH为8.0,能够重复利用4次以上。[结论]该研究为利用AChE快速检测蔬菜水果中有机磷农药残留提供了理论依据。
[Objective] The aim was to explore the optimum immobilized conditions of acetylcholinesterase on filter paper coated with chitosan.[Method] Acetylcholinesterase was immobilized on filter paper coated with chitosan using glutaraldehyde as cross-linking reagent,bovine serum albumin as protective agent.And the physical and chemical properties of the immobilized enzyme were studied.[Result] The optimal immobilized conditions were as follows： 1 ml solution was prepared with 50 μl 150 U/ml acetylcholinesterase solution,50 μl 5%（V/V） glutaraldehyde,100 μl 1%（W/W） BSA and 0.2 mol/L phosphate buffer（pH 8.0）,and then the filter paper coated with chitosan was immersed for 8 h at 4 ℃.The optimum temperature of the immobilized acetylcholinesterase was 37 ℃,the optimum pH was 8.0,and its could be reused four times.[Conclusion] The study provides a theory basis for rapid determination of organophosphorus pesticide residues in vegetables and fruits.
出处
《安徽农业科学》
CAS
2012年第23期11653-11655,共3页
Journal of Anhui Agricultural Sciences
基金
山东省自然科学基金项目(ZR2011DL008
Y2008D48)
关键词
滤纸壳聚糖膜
乙酰胆碱酯酶
固定化
农药残留
Filter paper coated with chitosan
Acetylcholinesterase
Immobilization
Pesticide residue
