多孔壳聚糖膜固定葡萄糖氧化酶活性的X射线微区分析

甲壳素(chitin)是无脊椎动物,特别是节肢动物,如虾、蟹及昆虫等的外骨骼重要组成部分,其学名为β(1,4)-2-乙酰氨基-2-脱氧-D-葡萄糖。壳聚糖是甲壳素脱乙酰基的衍生物,其结构上具有许多—NH2、—OH等反应基团,对蛋白质具有极高的亲和性,且具有生物相容性好、无毒、可生物降解等

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

The localization of the activity of immobilized glucose oxidase on chitosan membrane was studied by X-ray microanalysis.Ce(NO_3)_3 and glucose were selected as capture and substrate,respectively.The substrate was hydrolyzed by immobilized glucose oxidase to produce glucose acid and H_2O_2 in Tris-HCl buffer(pH 7.1).The H_2O_2 was captured by Ce(NO_3)_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.