基于液质联用技术的大豆主要致敏原蛋白检测及特异性肽段筛选方法研究  

Detection of Soybean Main Allergens and Specific Peptides Based on Liquid Chromatography-Mass Spectroscopy

作　　者：阴子悦 贾鑫 Yin Ziyue;Jia Xin(College of Food Science and Nutritional Engineering,China Agricultural University,Beijing 100083)

机构地区：[1]中国农业大学食品科学与营养工程学院,北京100083

出　　处：《中国粮油学报》2025年第1期193-201,共9页Journal of the Chinese Cereals and Oils Association

基　　金：山东省重点研发计划项目(2022CXGC010602)。

摘　　要：大豆[Glycine max(L.)Merr.]是一种重要的食品原料,同时富含致敏蛋白。目前,部分致敏原蛋白还未被确证,且致敏原蛋白的相对含量及其在大豆品种间的表达变异性尚不清楚。本研究利用高效液相色谱-串联质谱联用(HPLC-MS/MS)技术,以中国特色的种植范围较广的高蛋白大豆品种冀豆23(J23)、冀豆17(J17)和高油品种五星3(WX3)为研究对象,利用Orbitrap Fusion HPLC-MS/MS对其大豆蛋白进行检测和分析,建立多种致敏原蛋白及致敏特异性肽段的筛选方法。共确证了22中致敏蛋白组,51个致敏蛋白亚基和5923条致敏肽段。3个品种大豆总致敏蛋白含量差异不大,大豆球蛋白G1亚基在3个品种中占比最高,而WX3中存在特有的大豆球蛋白G2亚基以及β-伴大豆球蛋白α亚基,这可能是由于大豆品种的特异性导致的,而从J17中筛选的特异性肽段丰度较低。此外,3个大豆品种中的主要致敏蛋白β-伴大豆球蛋白β亚基和大豆球蛋白G1亚基中筛选得到的LITLAIPVNKPGR、VLIVPQNFVVAAR、VPSGTTYYVVNPDNNENLR等34条特异性肽段为首次报道,可作为检测高油酸、高蛋白大豆品种致敏原蛋白的特异性肽段。Soybean(Glycine max(L.)Merr.)is an important food material,rich in allergenic protein.At present,there are still soy allergens that have yet to be identified,also the relative amount of allergens among different soybean varieties remain unclear.In this study,Orbitrap Fusion HPLC-MS/MS was used to detect and analyze the extracted soybean protein from Chinese protein-rich soybean cultivars Jidou 23(J23)and Jidou 17(J17),as well as an oil-rich cultivar Wuxing 3(WX3).A total of 22 allergen groups,51 allergen protein subunits and 5923 allergen peptides were identified.There were few differences in the total allergenic protein content amount three varieties,with the G1 subunit of soybean globulin accounting for the highest proportion among the three varieties.While the globulin G2 subunit andβ-conglycininαsubunit were only identified in WX3 cultivar,which may be species specific,the specific peptide abundance identified from J17 was low.In addition,34 specific peptides,such as LITLAIPVNKPGR,VLIVPQNFVVAAR,VPSGTTYYVVVNPDNNLR,etc.identified in the main soy allergen proteins,such as globulin G1 subunit andβ-conglycininβsubunit,in three soybean varieties were reported for the first time,could be used as specific peptides for the detection of allergenic proteins from high-oleic acid and high-protein soybean varieties.

关 键 词：大豆致敏蛋白 致敏性 高效液相色谱-串联质谱联用 

分 类 号：TS214.2[轻工技术与工程—粮食、油脂及植物蛋白工程]