卢知浩，程青丽，冯晓文，赵晓涵，谷瑞增，鲁军，刘文颖*，李国明*
（中国食品发酵工业研究院有限公司，北京市蛋白功能肽工程技术研究中心，北京 100015）
摘要：目的：牡蛎存在致敏性是影响其作为饮料原料发展的主要原因，牡蛎中的肌浆钙结合蛋白（sarcoplasmiccalcium-binding protein，SCP）在细胞和肌肉功能中起着至关重要的作用，已被确定为牡蛎中的过敏原。为研究该类过敏原的具体致敏性原理，对其线性表位进行鉴定。方法：以太平洋牡蛎（Crassostrea gigas）的SCP 氨基酸序列为基础，制备并鉴定了分子量为21 kDa 的重组SCP（rSCP），并作为后续实验材料，建立了动物致敏模型进行实验。结果：rSCP 与大鼠血清具有较强的IgE 结合活性，过敏反应能引起大鼠脏器发生病变。最重要的是，五个表位肽首次通过间接竞争酶联免疫吸附实验（indirect competitive enzyme-linked immunosorbent assay，icELISA）鉴定为线性免疫显性表位。结论：线性表位研究对于理解人类对牡蛎的超敏反应具有重要意义，可为判断牡蛎蛋白饮料的致敏性奠定理论基础，并有助于提供有效的预防措施和治疗方案，促进该类饮料产品的发展。
关键词：太平洋牡蛎；重组肌浆钙结合蛋白；线性表位；过敏原
Prokaryotic Expression and Linear Epitope Identification of Potential Allergenic Proteinsin OysterProtein Beverages
LU Zhi-hao，CHENG Qing-li，FENG Xiao-wen，ZHAO Xiao-han，GU Rui-zeng，LU Jun，LIU Wen-ying*，LI Guo-ming*
（China National Research Institute of Food and Fermentation Industries Co. Ltd.，Beijing Engineering Research Centerof Protein and Functional Peptides，Beijing 100015，China）
Abstract: Purpose：The allergenicity of oysters is the main reason that affects its development as a beverage rawmaterial. The sarcoplasmic calcium- binding protein （SCP） in oysters plays a crucial role in cellular and musclefunction and was identified as an allergen in oysters. To study the specific sensitization mechanism of this allergen，its linear epitopes were identified.Methods：In this study，recombinant SCP（rSCP）with a molecular mass of 21 kDawas produced and identified based on the SCP amino acid sequence of pacific oyster（Crassostrea gigas），and wasused as a follow-up experimental material. We established an animal sensitization model in the study.Results：rSCPdisplayed stronger IgE-binding activity with rats serum，and allergic reaction could cause pathological changes in ratorgans. Most importantly，five epitope peptides were identified as linear immunodominant epitopes by indirectcompetitive enzyme- linked immunosorbent assay （icELISA） for the first time. Conclusion： These results aresignificant for understanding the hypersensitization of humans to oysters and offer available preventive measures and treatment programs in further research. These results are significant for understanding the hypersensitization ofhumans to oysters and can lay a theoretical basis for judging the allergenicity of oyster protein beverages. Andprovide effective preventive measures and treatment plans to promote the development of beverages.
Key words: crassostrea gigas；recombinant sarcoplasmic calcium-binding protein；linear epitopes；allergen
中图分类号：TS201.6 文献标志码：A 文章编号：1007-7871（2023）03-0011-08
2023，Vol.26，No.3