大豆β-伴球蛋白脱糖基化及其酶解后抗原活性变化  

Deglycosylation of Soybeanβ-Conglycinin and Changes in Its Antigenic Activity After Enzymatic Hydrolysis

作　　者：王章存 刘洋 安广杰 胡金强 赵学伟 贺志铮 Wang Zhangcun;Liu Yang;An Guangjie;Hu Jinqiang;Zhao Xuewei;He Zhizheng(School of Food and Bioengineering,Zhengzhou University of Light Industry,Zhengzhou 450001;Henan International Joint Laboratory of Food Safety,Zhengzhou 450001)

机构地区：[1]郑州轻工业大学食品与生物工程学院,郑州450001 [2]河南省食品安全国际联合实验室,郑州450001

出　　处：《中国粮油学报》2020年第12期62-66,共5页Journal of the Chinese Cereals and Oils Association

基　　金：国家自然科学基金(31671782)。

摘　　要：大豆蛋白的抗原性是影响其食用安全的重要因素。为了探讨大豆主要抗原蛋白β-伴球蛋白中的糖基化在其抗原反应中的地位和作用,本研究用N-糖苷酶PNGase F对β-伴球蛋白脱糖基化处理后,脱糖和未脱糖β-伴球蛋白在相同条件下用风味蛋白酶水解,用SDS-PAGE电泳和专用β-伴球蛋白酶联免疫试剂盒分别测定了脱糖和未脱糖β-伴球蛋白在酶解过程中的蛋白质组分和抗原性变化。结果表明,β-伴球蛋白分子中不仅含有N-糖肽键连接,也有O-糖肽键连接。脱糖后β-伴球蛋白抗原活性降低到原来的60.1%;脱糖和不脱糖两种β-伴球蛋白及其酶解产物电泳谱带和抗原性变化均有显著差异。酶解180 min时,β-伴球蛋白抗原活性下降为43.4%,脱糖β-伴球蛋白抗原活性仅剩25.0%。此结果揭示了糖链存在影响β-伴球蛋白抗原活性和酶解时的降解行为,为探讨有效消除大豆蛋白抗原性提供一定的理论基础。The edible safety of soybean protein is greatly affected by its antigenicity.In order to study the role of glycosylation in the antigenic reaction ofβ-conglycinin,an important antigenic protein,the N-glycosidase PNGase F was used to remove the N-linked glycopeptide of the protein.Theβ-conglycinin and its deglycosylated form were hydrolyzed with Flavoryzme under same condition.SDS-PAGE and specialβ-conglycinin ELISA kit were used respectively to analyze the components changes and the antigenicity changes of the two form proteins during the hydrolysis.The results showed that there were not only N-linked,but also O-linked glycopeptide inβ-conglycinin.The antigenic activity of deglycosylatedβ-conglycinin was only 60.1%of the originalβ-conglycinin.The electrophoretic profiles between the hydrolysates ofβ-conglycinin and its deglycosylated form were significantly different.After 180 minutes of enzymolysis with Flavoryzme,the antigenic activity ofβ-conglycinin and its deglycosylated form were reduced to 43.4%and 25.0%,respectively.The results revealed that The activity and the degradation behavior ofβ-conglycinin were affected by the existence of glycosylation during enzymolysis,which provided a theoretical basis for effectively eliminating the antigenicity of soybean protein.

关 键 词：大豆β-伴球蛋白 脱糖基化 酶解 抗原活性 电泳 

分 类 号：TS214.2[轻工技术与工程—粮食、油脂及植物蛋白工程]