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粉尘螨1类变应原的酶解工艺优化及其过敏原性检测
  • ISSN号:1005-9903
  • 期刊名称:中国实验方剂学杂志
  • 时间:2012
  • 页码:8-12
  • 分类:R283.6[医药卫生—中药学;医药卫生—中医学]
  • 作者机构:[1]皖南医学院医学寄生虫学教研室,安徽芜湖241002
  • 相关基金:国家自然科学基金项目(30872367;81172790)
  • 相关项目:微小RNA(miRNA)对实验性哮喘CD4+ T细胞增殖和分化的调节机制研究
中文摘要:

目的:优化粉尘螨1类变应原proDerf1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proDerf1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proDerf1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pH6.5,温度60℃,水解时间4h,酶用量4000U·g-1;胰蛋白酶最佳水解条件为pH8.0,温度45℃,水解时间4h,酶用量5000U·g-1;ELISA检测表明与proDerf1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proDerf1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。

英文摘要:

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.

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期刊信息
  • 《中国实验方剂学杂志》
  • 北大核心期刊(2011版)
  • 主管单位:国家中医药管理局
  • 主办单位:中国中医科学院中药研究所 中华中医药学会
  • 主编:吴以岭
  • 地址:北京东直门内南小街16号中国中医科学院中药研究所304室
  • 邮编:100700
  • 邮箱:syfjx_2010@188.com
  • 电话:010-84076882
  • 国际标准刊号:ISSN:1005-9903
  • 国内统一刊号:ISSN:11-3495/R
  • 邮发代号:2-417
  • 获奖情况:
  • 全国中医药优秀期刊三等奖,首届《CAJ-CD规范》执行优秀奖
  • 国内外数据库收录:
  • 美国化学文摘(网络版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版)
  • 被引量:59476