糖基化修饰消减南极磷虾原肌球蛋白致敏性研究  

Allergenicity Reduction of Antarctic Krill(Euphausia superba)Tropomyosin through Glycosylation Modification

作　　者：张书琪 王舒雅 贾淑琪 刘柯欣 孙娜[1] ZHANG Shuqi;WANG Shuya;JIA Shuqi;LIU Kexin;SUN Na(School of Food Science and Technology,SKL of Marine Food Processing&Safety Control,National Engineering Research Center of Seafood,Food Engineering Research Center,Dalian Polytechnic University,Dalian 116034,China)

机构地区：[1]大连工业大学食品学院,海洋食品加工与安全控制全国重点实验室,国家海洋食品工程技术研究中心,食品工程技术研究中心,辽宁大连116034

出　　处：《食品工业科技》2025年第8期9-22,共14页Science and Technology of Food Industry

基　　金：大连市科技人才创新支持计划项目(2022RY04)。

摘　　要：南极磷虾是优质的蛋白质资源,然而其主要过敏原原肌球蛋白的强致敏性仍是亟待解决的问题。为了消减南极磷虾原肌球蛋白的致敏性,本研究用单糖(核糖、葡萄糖)和低聚糖(低聚甘露糖、低聚半乳糖)对南极磷虾原肌球蛋白进行糖基化处理。结果显示,糖基化后原肌球蛋白的分子量增加,自由氨基含量降低,二级结构从α-螺旋转变为β-折叠,同时由于糖基化处理促使原肌球蛋白中的芳香族氨基酸残基暴露在疏水环境中,导致荧光强度降低,这表明原肌球蛋白与四种糖发生了糖基化反应;进一步采用液相色谱质谱联用技术探究了原肌球蛋白的糖基化程度及其过敏原表位的修饰情况,结果发现,单糖糖基化修饰原肌球蛋白的糖基化程度显著高于低聚糖(P<0.05),同时低聚甘露糖和低聚半乳糖糖基化修饰原肌球蛋白,位于其过敏原表位上的糖基化位点数量分别为9、11个,高于核糖(8个)和葡萄糖(8个)的糖基化位点数量;BALB/c小鼠致敏模型结果表明,与受试原肌球蛋白的小鼠相比,受试糖基化原肌球蛋白小鼠的过敏症状减轻,包括较低水平的IgE、IgG1、IL-4、肥大细胞脱颗粒率、组胺以及血管和肠壁通透性,同时受试低聚半乳糖糖基化处理的原肌球蛋白小鼠的症状减轻程度更加明显,可能是由于低聚半乳糖对原肌球蛋白过敏原表位的修饰数量较多。本研究为南极磷虾原肌球蛋白致敏性的消减及其相关低致敏性产品的开发提供了理论基础。Antarctic krill(Euphausia superba)was a premium protein resource.However,its strong allergenicity remained a problem that awaited resolution.To reduce the allergenicity of Antarctic krill tropomyosin,monosaccharides(ribose,glucose)and oligosaccharides(mannooligosaccharide,galactooligosaccharide)were used for glycosylation.Results showed that glycosylated tropomyosin underwent a decrease in free amino groups and an increase in molecular weight.Meanwhile,the secondary structure experienced a transition fromα-helix toβ-sheet,the exposure of aromatic amino acid residues to a hydrophobic environment resulted in decreased fluorescence intensity of glycosylated tropomyosin.These findings demonstrated that four kinds of sugars successfully glycosylation with tropomyosin.The glycosylation degree of tropomyosin and the modification of its allergenic epitopes were further investigated using liquid chromatography-tandem MS.Results revealed that the glycosylation degree of monosaccharide glycosylated tropomyosin significantly higher than that of oligosaccharide glycosylated tropomyosin(P<0.05),and that the number of glycosylation sites on allergenic epitopes of tropomyosin modified by mannooligosaccharide and galactooligosaccharide was 9 and 11,respectively,higher than those of tropomyosin modified by ribose and glucose,which showed 8 and 8,respectively.Results from the BALB/c mice model showed that the mice gavaged with glycosylated tropomyosin exhibited reduced allergic responses compared to those gavaged with tropomyosin,including lower levels of IgE,IgG1,IL-4,mast cell degranulation,histamine,vascular permeability,and intestinal permeability.It was worth noting that the mice gavaged with galactooligosaccharideglycosylated tropomyosin showed a more pronounced reduction in allergic responses,which was attributed to the fact that galactooligosaccharide modified more allergen epitopes of tropomyosin than ribose,glucose,and mannooligosaccharide.This study provided a theoretical basis for reducing the allergenicity of Antarct

关 键 词：南极磷虾 原肌球蛋白 致敏性 糖基化位点 过敏原表位 

分 类 号：TS201.6[轻工技术与工程—食品科学]