摘要
为研究小麦非麸质致敏原α-淀粉酶抑制剂的表位定位及消减技术,通过生物信息学预测工具DNAstar、IMED、IEDB对其线性表位进行预测,综合分析得出6条候选表位,利用间接竞争酶联免疫吸附实验对表位进行验证,得到4条线性表位。利用多酚与致敏原互作消减致敏,采用非共价方法制得α-淀粉酶抑制剂-表没子儿茶素没食子酸酯(epigallocatechin gallate,EGCG)/表没食子酸儿茶素复合物,通过荧光光谱、圆二色谱、紫外光谱进行表征,利用间接竞争酶联免疫吸附实验对消减效果进行研究,结果表明α-淀粉酶抑制剂与EGCG物质的量比为1∶30时,复合物的IgE结合水平达到最低,结合能力下降22.7%。通过Autodock软件进行分子对接模拟,经与表位进行比对,发现EGCG可与α-淀粉酶抑制剂线性表位及其附近残基相互作用从而达到致敏性消减的效果。研究结果可为小麦非麸质致敏原表位定位及低敏互作物的制备提供参考。
This study aimed at the epitope mapping and allergencity reduction of the wheat non-gluten allergenα-amylase inhibitor.Its linear epitopes were predicted by using bioinformatic prediction tools DNAstar,IMED,and IEDB.Six candidate epitopes were obtained by comprehensive analysis,and four linear epitopes were verified by using indirect competition enzyme-linked immunosorbent assay(ic-ELISA).The allergenicity of the allergen was reduced by interaction with polyphenols,A non-covalent complex ofα-amylase inhibitor and epigallocatechin gallate(EGCG)was prepared and characterized by fluorescence spectroscopy,circular dichroism(CD)spectroscopy and ultraviolet(UV)spectroscopy,and the efficiency of allergenicity reduction was investigated by ic-ELISA.Results showed that the complex with a molar ratio ofα-amylase inhibitor to EGCG of 1:30 had the lowest IgE-binding capacity.The binding capacity was reduced by 22.7%.Molecular docking simulations by using Autodock software further demonstrated that the allergenicity reduction was due to the interactions of the linear epitopes ofα-amylase inhibitor and their nearby residues with EGCG.This study can provide a reference for the epitope mapping of wheat non-gluten allergens and the preparation of polyphenols to reduce their allergenicity.
作者
王垚
张巧智
王彦波
傅玲琳
WANG Yao;ZHANG Qiaozhi;WANG Yanbo;FU Linglin(School of Food Science and Biological Engineering,Zhejiang Research Institute of Food Quality and Safety Engineering,Zhejiang Gongshang University,Hangzhou 310018,China)
出处
《食品科学》
EI
CAS
CSCD
北大核心
2023年第22期200-210,共11页
Food Science
基金
浙江省基础公益研究项目(LGN22C200027)
浙江省“三农九方”农业科技协作计划揭榜挂帅项目(2022SNJF078)
浙江工商大学“数字+”学科建设重大项目(SZJ2022A010)。
关键词
食物过敏
小麦致敏原
表位定位
分子互作
致敏原消减
food allergy
wheat allergens
epitope mapping
intermolecular interaction
allergenicity reduction of allergens
