杨君扬,张烘煜,陈忠琴,谭明堂,林海生,高加龙,郑惠娜,曹文红,杨维,陈忆宾

• 1:广东海洋大学食品科技学院
• 2:国家贝类加工技术研发分中心(湛江)
• 3:海南翔泰渔业股份有限公司

摘要(Abstract)：

【目的】通过螯合结构优化实现牡蛎肽硒复合物免疫活性功能的提升。【方法】借助Capto Q离子交换层析对牡蛎肽及其硒螯合物进行分离纯化，筛选出易与硒结合的肽；通过红外和圆二色光谱对比螯合前后官能团的变化确定硒的结合位点；运用液相色谱-串联质谱联用（LC-MS/MS）和分子对接对硒螯合肽结构特征及结合位点进行分析；以RAW264.7巨噬细胞为实验对象，对比牡蛎肽及其硒螯合物洗脱组分（P2和F2）的细胞免疫应答变化。【结果】波谱和分子对接结果均表明，肽链上的氨基、羧基、羰基等均参与硒离子螯合过程的共价反应，且螯合反应使肽链的二级结构从无规则趋向规则的折叠、转角结构；LC-MS/MS结果表明，在所检测出的螯合物中，相对分子质量越大，碱性氨基酸占比和等电点（PI）值越高的肽链具有更强的硒螯合能力。体外细胞实验表明，F2能显著提高RAW264.7细胞一氧化氮（NO）、活性氧（ROS）、肿瘤坏死因子-α(TNF-α)、白细胞介素（IL-6）的分泌；增加巨噬细胞的吞噬活性；促进免疫相关蛋白COX-2和iNOS的表达，且效果显著优于P2。【结论】牡蛎肽在与硒螯合后结构发生变化，PI值越大的肽更易于与硒发生螯合，螯合后免疫活性得到显著增强。

关键词(KeyWords)： 牡蛎肽;硒螯合物;特征肽;RAW264.7细胞;免疫活性

Abstract:

Keywords:

基金项目(Foundation): 国家贝类产业技术体系（CARS-49）;; 国家重点研发计划课题（2018YFD0901105）

作者(Author): 杨君扬,张烘煜,陈忠琴,谭明堂,林海生,高加龙,郑惠娜,曹文红,杨维,陈忆宾

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