食品与发酵工业 >

2026 , Vol. 52 >Issue 12: 61 - 69

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.044437

研究报告

核桃内种皮多酚提取物的共价修饰对核桃球蛋白抗原性、结构及过敏原蛋白质谱相对丰度的影响

  • 任丽秋 ,
  • 张燕 ,
  • 王杰 ,
  • 陆乃彦 ,
  • 成向荣
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  • (江南大学 食品学院,江苏 无锡,214122)
第一作者:硕士研究生(成向荣副教授为通信作者,E-mail:cheng-xiangrong@hotmail.com)

收稿日期: 2025-09-05

  修回日期: 2025-11-06

  网络出版日期: 2026-07-17

基金资助

国家自然科学基金项目(82103836)

收起

Effect of covalent modification by polyphenol extract from walnut pellicle on the antigenicity, structure, and relative abundance of allergen protein profiles of walnut globulin

  • REN Liqiu ,
  • ZHANG Yan ,
  • WANG Jie ,
  • LU Naiyan ,
  • CHENG Xiangrong
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  • (School of Food Science and Technology, Jiangnan University, Wuxi 214122, China)

Received date: 2025-09-05

  Revised date: 2025-11-06

  Online published: 2026-07-17

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摘要

核桃是我国的传统坚果,但核桃蛋白质中含有8种过敏原(Jug r 1~Jug r 8)。该研究旨在探究核桃内种皮多酚提取物(polyphenol extract from walnut pellicle,PEWP)对核桃球蛋白(walnut globulin,WG)的抗原性的影响及其与蛋白质结构和过敏原蛋白质谱相对丰度变化的关系。通过碱法制备WG-PEWP复合物,采用光谱技术及体外免疫实验评估PEWP对WG结构和抗原性的影响,同时运用蛋白质组学技术进一步探究复合物中核桃过敏原的变化。结果显示,PEWP的修饰改变了WG的二级与三级结构。随着PEWP添加量的增加,WG的免疫球蛋白G(immunoglobulin G,IgG)结合能力从(72.68±2.68)%显著降低至(43.26±0.25)%(P<0.05)。相关性分析表明复合物的IgG结合能力与其结构变化存在统计学显著相关性(P<0.05)。蛋白质组学分析结果显示WG中含有6种核桃过敏原,经PEWP修饰后,4种过敏原蛋白(Jug r 2、Jug r 4、Jug r 6和Jug r 8)的质谱相对丰度显著降低。复合物的酸水解产物分析发现鞣花酸(ellagic acid,EA)是PEWP中与WG发生相互作用的主要多酚成分。这些结果表明,PEWP的添加降低了WG的结构有序性和抗原性,而PEWP中的EA可能通过共价修饰核桃过敏原蛋白来减弱其抗原性。该研究旨在为开发低致敏性核桃产品提供理论基础及技术层面的创新见解。

关键词： 核桃过敏; 核桃球蛋白; 蛋白质组学; 蛋白质结构; 蛋白质-多酚复合物

本文引用格式

任丽秋 , 张燕 , 王杰 , 陆乃彦 , 成向荣 . 核桃内种皮多酚提取物的共价修饰对核桃球蛋白抗原性、结构及过敏原蛋白质谱相对丰度的影响[J]. 食品与发酵工业, 2026 , 52(12) : 61 -69 . DOI: 10.13995/j.cnki.11-1802/ts.044437

Abstract

Walnuts are traditional nuts in China, but the proteins in walnuts contain eight types of allergens (Jug r 1-Jug r 8).This study aimed to investigate the effect of polyphenol extract from walnut pellicle (PEWP) on the antigenicity of walnut globulin (WG), as well as its relationship with changes in protein structure and the relative abundance of allergen protein profiles.WG-PEWP complexes were prepared via alkaline extraction.Spectral techniques and in vitro immunological assays assessed the effects of PEWP on WG’s structure and antigenicity.Meanwhile, proteomic analysis further explored changes in walnut allergens within the complexes.Results showed that PEWP modification altered the secondary and tertiary structures of WG.As PEWP addition increased, WG’s IgG-binding capacity decreased significantly from (72.68±2.68)% to (43.26±0.25)% (P<0.05).Correlation analysis revealed a statistically significant correlation between the IgG-binding capacity of the complexes and their structural changes (P<0.05).Proteomic analysis identified six walnut allergens in WG.After PEWP modification, the mass spectrometric relative abundances of four allergens (Jug r 2, Jug r 4, Jug r 6, and Jug r 8) decreased significantly.Analysis of the acid hydrolysates of the complexes identified ellagic acid (EA) as the main polyphenolic component in PEWP that interacts with WG.These results indicate that PEWP addition reduces the structural order and antigenicity of WG.EA in PEWP may weaken allergenicity by covalently modifying walnut allergenic proteins.This study provides a theoretical basis and technical insights for developing hypoallergenic walnut products.

Key words： walnut allergy; walnut globulin; proteomics; protein structure; protein-polyphenol complex

参考文献

[1] LYONS S A, DATEMA M R, LE T M, et al.Walnut allergy across Europe:Distribution of allergen sensitization patterns and prediction of severity[J].The Journal of Allergy and Clinical Immunology:In Practice, 2021, 9(1):225-235.e10.
[2] SPOLIDORO G C I, LISIK D, NYASSI S, et al.Prevalence of tree nut allergy in Europe:A systematic review and meta‐analysis[J].Allergy, 2024, 79(2):302-323.
[3] LEUNG A S, PACHARN P, TANGVALELERD S, et al.Food allergy in a changing dietary landscape:A focus on the Asia Pacific region[J].Pediatric Allergy and Immunology, 2024, 35(8):e14211.
[4] 姜松松, 赵博, 韩诗雯, 等.不同热加工方式对核桃蛋白致敏性的影响[J].食品科学, 2018, 39(13):94-99.
JIANG S S, ZHAO B, HAN S W, et al.Effect of different thermal processing treatments on allergenicity of walnut proteins[J].Food Science, 2018, 39(13):94-99.
[5] MANZOOR M, TCHAMENI Z F N, BHAT Z F, et al.Recent insights on the conformational changes, functionality, and physiological properties of plant-based protein-polyphenol conjugates[J].Food and Bioprocess Technology, 2024, 17(8):2131-2154.
[6] XU X Q, CAO Q, YUAN J, et al.Shedding light on the interaction of ovalbumin and resveratrol:Structure, digestibility, transport, and allergenicity assessment of OVA-RES complexes[J].Journal of the Science of Food and Agriculture, 2024, 104(3):1645-1655.
[7] VICTORIA GIL M, GUTIÉRREZ-DÍAZ G, HIGUERO N, et al.Targeting cow’s milk allergy using hypoallergenic protein-polyphenol formulas:A proof of concept[J].Food Chemistry, 2025, 463(Pt 3):141285.
[8] AFZAAL M, SAEED F, HUSSAIN M, et al.Proteomics as a promising biomarker in food authentication, quality and safety:A review[J].Food Science & Nutrition, 2022, 10(7):2333-2346.
[9] CARRERA M, CAÑAS B, GALLARDO J M.Advanced proteomics and systems biology applied to study food allergy[J].Current Opinion in Food Science, 2018, 22:9-16.
[10] MA J H, REN L Q, TANG T X, et al.Effects of polyphenols from walnut pellicle on the structure and allergenicity of walnut globulin[J].Food Bioscience, 2024, 60:104381.
[11] 马家辉. 核桃蛋白与内种皮多酚相互作用及其对蛋白结构与致敏性的影响[D].无锡:江南大学, 2024.
MA J H.The interaction between walnut protein and pellicle polyphenols and its effects onprotein structure and allergenicity[D].Wuxi:Jiangnan University, 2024.
[12] JIA X D, XU M Y, TAN W Y, et al.Proteomic and transcriptomic analyses provide new insights into the mechanism underlying lipid deterioration in pecan kernels during storage[J].Journal of Agricultural and Food Chemistry, 2024, 72(17):10127-10137.
[13] CHENG X R, MA J H, AMADOU I, et al.Electrophilic components from Xiaoheiyao (rhizomes of in ula nervosa Wall.) alleviate the production of heterocyclic aromatic amines via creatinine inhibition[J].Food Chemistry, 2023, 404(Pt A):134561.
[14] QUAN T H, BENJAKUL S, SAE-LEAW T, et al.Protein-polyphenol conjugates:Antioxidant property, functionalities and their applications[J].Trends in Food Science & Technology, 2019, 91:507-517.
[15] BARBOSA B S T, ARAÚJO S C, CORDEIRO Y, et al.Impact of the covalent interaction between ferulic acid and ovalbumin on the structure and functional properties of the protein[J].Food Biophysics, 2024, 20(1):28.
[16] TANG M G, YANG M, XU L K, et al.Covalent interactions between walnut protein isolate and chlorogenic acid impact the formation, structure, functionality and emulsions stability of the isolates[J].LWT, 2024, 199:115991.
[17] LIU C, TANG P P, LIU X B, et al.Different polyphenols on cold plasma-induced covalent ovalbumin-polyphenols conjugates and affecting the antigenicity of OVA and its functionalities[J].Food Hydrocolloids, 2025, 162:110929.
[18] ZAINUDIN M A M, JONGBERG S, LUND M N.Combination of light and oxygen accelerates formation of covalent protein-polyphenol bonding during chill storage of meat added 4-methyl catechol[J].Food Chemistry, 2021, 334:127611.
[19] LI F, XIANG T J, JIANG L, et al.New insights into ultrasound-assisted noncovalent nano complexes of β-lactoglobulin and neochlorogenic acid/cryptochlorogenic acid and its potential application for curcumin loading[J].Food Research International, 2025, 199:115384.
[20] AHMAD ASHWAR B, GANI A.Noncovalent interactions of sea buckthorn polyphenols with casein and whey proteins:Effect on the stability, antioxidant potential, and bioaccessibility of polyphenols[J].ACS Food Science & Technology, 2021, 1(7):1206-1214.
[21] LIU L L, YANG J H, YU Y, et al.Elucidating interaction mechanisms of catechin, proanthocyanidin, and tea polyphenol with porcine serum albumin:Insights into functional properties and complex formation[J].Food Bioscience, 2024, 62:105324.
[22] ZHAO S N, DENG Y, YAN T Y, et al.Explore the interaction between ellagic acid and zein using multi-spectroscopy analysis and molecular docking[J].Foods, 2022, 11(18):2764.
[23] 张春子, 马小燕, 阿布里孜·阿不都热合曼, 等.矢车菊素-3-O-葡萄糖苷和矢车菊素对酪氨酸酶的抑制作用及其机理研究[J].食品与发酵工业, 2025, 51(10):224-232.
ZHANG C Z, MA X Y, ABRIZI A, et al.Inhibition effects and mechanisms of cyanidin-3-O-glicoside and cyanidin on tyrosinase[J].Food and Fermentation Industries, 2025, 51(10):224-232.
[24] WEI Z H, YANG W, FAN R, et al.Evaluation of structural and functional properties of protein-EGCG complexes and their ability of stabilizing a model β-carotene emulsion[J].Food Hydrocolloids, 2015, 45:337-350.
[25] CLARE MILLS E N, JENKINS J A, ALCOCER M J C, et al.Structural, biological, and evolutionary relationships of plant food allergens sensitizing via the gastrointestinal tract[J].Critical Reviews in Food Science and Nutrition, 2004, 44(5):379-407.
[26] MATTISON C P, RAI R, SETTLAGE R E, et al.RNA-seq analysis of developing pecan (Carya illinoinensis) embryos reveals parallel expression patterns among allergen and lipid metabolism genes[J].Journal of Agricultural and Food Chemistry, 2017, 65(7):1443-1455.
[27] KONG X Z, HUANG Z L, ZHANG C M, et al.Phenolic compounds in walnut pellicle improve walnut (Juglans regia L.) protein solubility under pH-shifting condition[J].Food Research International, 2023, 163:112156.
[28] WANG Y Z, CHEN Y, LV J, et al.Characterization of walnut protein isolate-polyphenol nanoconjugates for the developing a food-grade delivery system[J].Journal of Food Science and Technology, 2023, 60(9):2454-2467.
[29] ZHANG Y, LIU C Q, SU M N, et al.Effect of phenolics on amandin immunoreactivity[J].LWT, 2018, 98:515-523.
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