凡纳滨对虾肌质钙结合蛋白纯化鉴定、理化特性及抗原表位分析

doi:10.7506/spkx1002-6630-20231023-190

食品科学 ›› 2025, Vol. 46 ›› Issue (2): 30-37.doi: 10.7506/spkx1002-6630-20231023-190

凡纳滨对虾肌质钙结合蛋白纯化鉴定、理化特性及抗原表位分析

陈伟，周军君，陈雅纯，贾英民，马爱进

（北京工商大学食品与健康学院，北京 100048）

出版日期:2025-01-25 发布日期:2024-12-30
基金资助:
国家自然科学基金面上项目（31972184）

Purification, Physicochemical Properties, and Epitope Analysis of Sarcoplasmic Calcium-Binding Protein from Penaeus vannamei

CHEN Wei, ZHOU Junjun, CHEN Yachun, JIA Yingmin, MA Aijin

(School of Food and Health, Beijing Technology and Business University, Beijing 100048, China)

Online:2025-01-25 Published:2024-12-30

摘要/Abstract

摘要： 以凡纳滨对虾（Penaeus vannamei）为原料，分离纯化和鉴定过敏原肌质钙结合蛋白（sarcoplasmic-calcium-binding protein，SCP），对其理化性质（免疫活性、热稳定性、pH值稳定性及消化稳定性）、二级结构和抗原表位进行研究。结果表明：采用蛋白粗提、硫酸铵分级沉淀、阴离子交换层析等步骤纯化得到的蛋白分子质量为21.6 kDa，经鉴定，该蛋白为凡纳滨对虾SCP，肽段覆盖率达93.26%。凡纳滨对虾SCP具有较强的免疫活性，在热处理温度≥65 ℃和强酸强碱条件下免疫活性减弱；SCP对肠液消化具有较强的稳定性，而对胃液消化稳定性较差。凡纳滨对虾SCP的二级结构中α-螺旋、β-折叠、β-转角和无规卷曲的相对含量分别为26%、16.9%、17.5%、39.6%。利用生物信息学工具结合免疫学技术最终预测识别出8 条凡纳滨对虾SCP抗原表位。

关键词: 凡纳滨对虾；肌质钙结合蛋白；稳定性；抗原表位

Abstract: Sarcoplasmic-calcium-binding protein (SCP) was isolated, puriﬁed and identified from Penaeus vannamei. Meanwhile, its physicochemical properties (immunoactivity, thermal stability, pH stability and digestive stability), secondary structures and epitopes were studied. The results showed that the purified protein, which was obtained by successive steps of crude protein extraction, ammonium sulfate fractionation and anion exchange chromatography and exhibited a molecular mass of 21.6 kDa, was identified as SCP with a peptide coverage of 93.26%. SCP had strong immunoactivity, which decreased during heat treatment (≥ 65 ℃) and under strongly acidic and alkaline conditions. SCP had strong stability against intestinal fluid digestion, but poor stability against gastric fluid digestion. The secondary structure of SCP consisted of 26% α-helix, 16.9% β-sheet, 17.5% β-turn, and 39.6% random coil. Eight epitopes in SCP were predicted and identified using bioinformatics tools combined with immunological techniques.

Key words: Penaeus vannamei; sarcoplasmic calcium-binding protein; stability; epitope

中图分类号:

TS201.2

陈伟，周军君，陈雅纯，贾英民，马爱进. 凡纳滨对虾肌质钙结合蛋白纯化鉴定、理化特性及抗原表位分析[J]. 食品科学, 2025, 46(2): 30-37 https://doi.org/10.7506/spkx1002-6630-20231023-190

CHEN Wei, ZHOU Junjun, CHEN Yachun, JIA Yingmin, MA Aijin. Purification, Physicochemical Properties, and Epitope Analysis of Sarcoplasmic Calcium-Binding Protein from Penaeus vannamei[J]. FOOD SCIENCE, 2025, 46(2): 30-37 https://doi.org/10.7506/spkx1002-6630-20231023-190

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凡纳滨对虾肌质钙结合蛋白纯化鉴定、理化特性及抗原表位分析

Purification, Physicochemical Properties, and Epitope Analysis of Sarcoplasmic Calcium-Binding Protein from Penaeus vannamei

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