多孔壳聚糖膜固定葡萄糖氧化酶活性的X射线微区分析  

X-ray Microanalysis of the Activity of Immobilized Glucose Oxidase on Chitosan Membrane

作　　者：马晓莉[1] 姚子华[1] 高远[1] 

机构地区：[1]河北大学医学部,河北保定071002

出　　处：《分析测试学报》2006年第4期27-31,共5页Journal of Instrumental Analysis

基　　金：河北省科技厅攻关项目(05276441)

摘　　要：利用X射线微区分析方法,对壳聚糖固定葡萄糖氧化酶活性进行定位分析。筛选出能捕捉固定化葡萄糖氧化酶活性部位的捕捉剂Ce(NO3)3,以葡萄糖作为底物,在Tris-HC l缓冲溶液中,底物经固定化葡萄糖氧化酶催化产生葡萄糖酸和H2O2,后者和捕捉剂反应生成氧化氢氧化铈(Ce(OH)2OOH)沉淀,沉积在固定化葡萄糖氧化酶的催化活性部位。结果表明:Ce(NO3)3可用做壳聚糖固定化葡萄糖氧化酶的捕捉剂,并得到了活性定位的最佳实验条件,多孔壳聚糖膜孔隙率高,表面积大,载酶量高,葡萄糖氧化酶均匀分布在膜孔内外。The localization of the activity of immobilized glucose oxidase on chitosan membrane was studied by X-ray microanalysis. Ce（ NO3 ）3 and glucose were selected as capture and substrate, respectively. The substrate was hydrolyzed by immobilized glucose oxidase to produce glucose acid and H2O2 in Tris - HCI buffer（pH 7. 1 ）. The H2O2 was Captured by Ce（ NO3 ） 3 to form （ ce（OH） 2OOH） precipitate which was then deposited on the catalytic active site of immobilized glucose oxidase. It is shown that the method is practicable and reliable. The optimum conditions for the localization of activity of immobilized glucose oxidase was studied. The porous chitosan membrane possesses controlled pore sizes, thoroughfare pores, big surface area and more amount of glucose oxidase immobilized. The distribution of activated enzyme was uniform on the surface of the porous chitosan membrane.

关 键 词：X射线微区分析 固定化葡萄糖氧化酶 酶活性 多孔壳聚糖膜 

分 类 号：O766.3[理学—晶体学]