小麦中的过敏原在酱油酿造过程中的降解分析

Degradation analysis of wheat allergens during the soy sauce brewing process

中文关键词:  酱油  小麦  过敏蛋白  酿造过程  降解

英文关键词:soy sauce  wheat  allergy protein  brewing process  degradation

中文摘要:

      目的 探究低盐固态酱油酿造过程不同阶段小麦过敏原的变化。方法 采用实验室模拟方法酿造低盐固态酱油, 采集酿造过程中的原料小麦面粉和制曲阶段14、27、36和44 h时的样品, 发酵阶段5、10、17、24和30 d, 灭菌前和灭菌后12个样品, 用实验室制备的小麦蛋白的多克隆抗体、免疫球蛋白G以及从医院获得的小麦过敏病人血清免疫球蛋白E对样品进行十二烷基硫酸钠聚丙烯酰胺凝胶电泳、免疫印迹和酶联免疫测定不同阶段样品的免疫原性。结果 经制曲、发酵处理和加热灭菌处理后, 凝胶电泳图中小麦面粉中的蛋白条带逐渐减少甚至消失, 其中制曲阶段变化程度最大。小麦过敏蛋白中的主要组分麦醇溶蛋白和麦谷蛋白在制曲阶段后期免疫印迹条带逐渐消失, 在进入发酵阶段第5 d时已完全检测不到免疫印迹条带。酶联免疫实验结果表明, 与未处理小麦面粉样品相比, 制曲、发酵和灭菌等处理过程均会不同程度降低小麦过敏原的抗原性和过敏原性, 在进入发酵阶段的第24 d小麦过敏蛋白含量已低于20 mg/kg, 致敏性基本被消除。结论 在实验室模拟低盐固态酱油酿造中, 小麦过敏蛋白的抗原性和过敏原性在制曲、发酵和灭菌等过程均不同程度降低, 其中制曲阶段作用最大, 成品酱油中小麦的致敏性基本被消除。

英文摘要:

      Objective To explore the changes of wheat allergens in different stages of the low-salt solid soy sauce brewing process. Methods The low-salt solid soy sauce was brewed by a laboratory simulation method. The raw material wheat flour in the brewing process and samples in the preparation stages of 14, 27, 36 and 44 h, the fermentation stages of 5, 10, 17, 24 and 30 d, and the 12 samples before and after sterilization were collected. The samples in different stages were subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis, immunoblotting and enzyme-linked immunosorbent assay for immunogenicity, using polyclonal antibody of wheat protein prepared in the laboratory, immunoglobulin G and serum immunoglobulin E of wheat allergy patients obtained from a hospital. Results After preparation, fermentation and heating sterilization, the protein bands in wheat flour gradually decreased or even disappeared in the gel electrophoresis picture, and the degree of change was the largest in the preparation stage. The western blot bands of gliadin and glutenin, the main components of wheat anaphylaxis proteins, gradually disappeared in the late stage of fermentation stage, and the western blot bands were completely undetectable when entering the 5th day of fermentation stage. The results of enzyme-linked immunosorbent assay showed that compared with untreated wheat flour samples, the treatment processes such as preparation, fermentation and sterilization would reduce the antigenicity and allergenicity of wheat allergens to different degrees. The content of wheat anaphylaxis protein was less than 20 mg/kg in the 24th day of fermentation stage, and the allergenicity was basically eliminated. Conclusion In the laboratory simulation of low-salt solid soy sauce brewing, the antigenicity and allergenicity of wheat allergenic protein decreases in different degrees during preparation, fermentation and sterilization, among which the preparation stage playes the biggest role, and the allergenicity of wheat in the finished soy sauce is basically eliminated.