枯草芽孢杆菌发酵对花生蛋白抗原性的影响  

Effect of fermentation with Bacillus subtilis on antigenicity of peanut protein

作　　者：陈屹轩 布冠好[1] 杨趁仙[1] 辛颖[1] 段晓杰 李梦瑶 CHEN Yixuan;BU Guanhao;YANG Chenxian;XIN Ying;DUAN Xiaojie;LI Mengyao(College of Food Science and Engineering,Henan University of Technology,Zhengzhou 450001,China)

机构地区：[1]河南工业大学粮油食品学院,河南郑州450001

出　　处：《河南工业大学学报(自然科学版)》2025年第1期19-25,32,共8页Journal of Henan University of Technology:Natural Science Edition

基　　金：国家自然科学基金项目(32472289,31871748);河南省自然科学基金面上项目(242300421317);河南省高等学校青年骨干教师培养计划(2019GGJS084);河南工业大学青年骨干教师培育计划(21420064);中国博士后科学基金面上项目(2021M701112);河南工业大学国家自然科学基金培育项目支持计划(2024PYJH003)。

摘　　要：为了探究枯草芽孢杆菌固态发酵降低花生蛋白抗原性的机理,采用间接竞争ELISA法测定蛋白抗原性、茚三酮比色法测定蛋白水解度、SDS-PAGE测定蛋白分子量,利用红外、紫外、荧光光谱分析花生蛋白结构变化对其抗原性的影响。结果表明:枯草芽孢杆菌固态发酵能够使花生蛋白发生显著降解,抗原性降低了67.61%,花生蛋白中主要过敏原Ara h 3抗原性降低了55.03%;发酵72 h花生蛋白水解度为9.40%,分子量小于15 kDa的蛋白含量显著增加;发酵后花生蛋白二级结构相对含量发生变化,α-螺旋和β-转角含量降低;蛋白结构逐渐展开,发酵48 h后微生物酶解作用减弱且部分蛋白发生聚合。枯草芽孢杆菌固态发酵可以通过改变花生蛋白结构破坏抗原表位,从而降低花生蛋白抗原性。To investigate the mechanism by which solid-state fermentation with Bacillus subtilis reduces the antigenicity of peanut protein,indirect competitive ELISA was employed to determine protein antigenicity,the ninhydrin colorimetric method was used to measure protein hydrolysis,and SDS-PAGE was utilized to assess protein molecular weight.Infrared spectrum,ultraviolet spectrum and endogenous fluorescence spectrum were applied to analyze the effects of structural changes in peanut protein on its antigenicity.Exogenous fluorescence spectroscopy was applied to analyze the surface hydrophobicity of peanut protein.The results demonstrated that solid-state fermentation with Bacillus subtilis significantly hydrolyzed peanut protein,resulting in a 67.6%reduction in antigenicity and a 55.03%decrease in the antigenicity of the major allergen Ara h 3 in peanut protein.After 72 hours of fermentation,the protein hydrolysis degree of peanut protein reached 9.40%.Major allergens in peanut protein are degraded,and the protein content with a molecular weight less than 15 kDa increased significantly.After fermentation,changes were observed in the relative content of secondary structure of peanut protein,with the relative contents ofα-helix andβ-turn decreased by 5.52%and 8.17%,respectively.The surface hydrophobicity of peanut protein gradually increased before 48 hours of fermentation,and after 48 hours of fermentation,the surface hydrophobicity of peanut protein gradually decreased with the fermentation time.In the early stage of fermentation,the tertiary structure of peanut protein changed,and the spatial structure gradually unfolded.But after 48 hours of fermentation,the microbial enzymatic hydrolysis weakened,accompanied by partial protein aggregation.Solid-state fermentation with Bacillus subtilis can disrupt antigenic epitopes by altering the structure of peanut protein,thereby reducing its antigenicity.

关 键 词：枯草芽孢杆菌 发酵 花生蛋白 抗原性 蛋白结构 

分 类 号：TS201.2[轻工技术与工程—食品科学]