粉尘螨1类变应原的酶解工艺优化及其过敏原性检测  

Optimization of Enzymatic Hydrolysis Technology for Dust Mite Group 1 Allergen and Its Allergenic Detection

作　　者：李秋雨[1] 姜玉新[1] 李朝品[1] 

机构地区：[1]皖南医学院医学寄生虫学教研室,安徽芜湖241002

出　　处：《中国实验方剂学杂志》2012年第22期8-12,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(30872367;81172790)

摘　　要：目的:优化粉尘螨1类变应原proDer f 1的酶解条件及探讨酶解产物的变应原性。方法:以水解度为指标,采用正交试验考察木瓜蛋白酶和胰蛋白酶水解proDer f 1蛋白的最佳工艺;采用间接酶联免疫吸附试验(ELISA)检测酶解产物的过敏原性。结果:温度是影响proDer f 1蛋白水解效果的主要因素,其中木瓜蛋白酶最佳水解条件为pH 6.5,温度60℃,水解时间4 h,酶用量4 000 U.g-1;胰蛋白酶最佳水解条件为pH 8.0,温度45℃,水解时间4 h,酶用量5 000 U.g-1;ELISA检测表明与proDer f 1变应原相比,木瓜蛋白酶水解产物和胰蛋白酶水解产物的过敏原性均显著减低,且胰蛋白酶水解产物的过敏原性较木瓜蛋白酶水解产物的过敏原性低。结论:优选的粉尘螨1类变应原proDer f 1蛋白的酶解工艺可行,且变应原酶解后,过敏原性有不同程度减低,尤以胰酶水解产物变化最为显著。Objective： To optimize enzymatic hydrolysis conditions of proDer f 1, and investigate allergenicity of enzymatic hydrolysate. Method： With hydrolysis degree as index, optimum hydrolysis technology of proDer f 1 was investigated by orthogonal test using papain and trypsin; Allergenicity of enzymatic hydrolysate was tested by ELISA. Result： Temperature was main factor affecting hydrolysis effect of proDer f 1, optimal hydrolysis conditions of proDer f 1 digested by papain were as follows： pH 6.5, temperature 60℃, hydrolysis time 4 h, the amount of papain 4 000 U-g^-1 ; Optimum hydrolysis conditions of trypsin was pH 8.0, temperature 45℃ , hydrolysis time 4 h, the amount of enzyme 5 000 U-g^-1 Compared with proDer f 1 allergen, ELISA analysis showed that allergens were reduced significantly of papain hydrolysate and trypsin hydrolysate, allergen of trypsin hydrolysate was lower than papain hydrolysate. Conclusion： Optimized enzymatic hydrolysis process of proDer f 1 protein was feasible, and after enzymatic hydrolysis of allergen, there was different degrees reduction of allergenicity, especially change of trypsin hydrolysate had the most significant.

关 键 词：proDer F 1蛋白 过敏原 蛋白酶水解 酶联免疫吸附试验 

分 类 号：R283.6[医药卫生—中药学]