马尾藻多酚的提取纯化工艺优化及其对虾原肌球蛋白致敏性的消减作用研究

Optimization of extraction and purification of polyphenol from Sargassum carpophyllum and its attenuating effects on the allergenicity of shrimp tropomyosin

中文关键词:  马尾藻多酚  提取纯化  原肌球蛋白  致敏性  抗过敏

英文关键词:Sargassum carpophyllum polyphenol  extraction and purification  tropomyosin  allergenicity  antiallergy

中文摘要:

      目的 研究马尾藻(Sargassum carpophyllum)多酚的提取纯化及其对虾原肌球蛋白致敏性的消减作用。方法 比较了马尾藻和其他5种海藻中多酚的含量, 通过透明质酸酶抑制活性实验确定出海藻多酚含量且透明质酸酶抑制活性最高的马尾藻为原材料, 并通过单因素优化和响应面优化确定了最佳的马尾藻多酚的提取条件, 进一步通过大孔树脂和高效液相色谱进行分离纯化。利用间接竞争酶联免疫吸附试验(enzyme linked immunosorbent assay, ELISA)和RBL-2H3肥大细胞模型解析马尾藻多酚对虾原肌球蛋白致敏性的消减作用。结果 马尾藻多酚的最佳提取工艺为: 提取温度为40℃, 提取时间为10 h, 马尾藻粉末:75%乙醇为1:16 (m/V), 马尾藻多酚提取得率可达18.18 mg/g, 经纯化后获得纯度为71.93%的马尾藻多酚, 其中含有水杨酸一水合物、3,4-二羟基-5-磺氧基苯甲酸和没食子酸。马尾藻多酚可通过结合原肌球蛋白中的芳香族氨基酸和荧光氨基酸, 遮蔽和破坏原肌球蛋白的抗原表位, 从而消减虾原肌球蛋白的致敏性。结论 本研究优化了马尾藻多酚提取纯化工艺, 得到的马尾藻多酚可以显著消减虾原肌球蛋白的致敏性, 表现出较强的抗过敏活性。

英文摘要:

      Objective To study the extraction and purification of Sargassum carpophyllum polyphenol and its attenuating effect on the allergencity of shrimp tropomyosin. Methods The content of polyphenols in Sargassum carpophyllum was compared with those in other 5 species of seaweed. The Sargassum carpophyllum with the highest content of polyphenols and the highest inhibitory activity of hyaluronidase were identified by hyaluronidase inhibitory activity experiment as the raw material. The optimal extraction conditions of Sargassum carpophyllum polyphenols were determined by single factor optimization and response surface optimization, and further separated and purified by macroporous resin and high performance liquid chromatography. Enzyme-linked immunosorbent assay (ELISA) and RBL-2H3 mast cell model were used to determine the attenuating effect of Sargassum carpophyllum polyphenol on the allergenicity of shrimp tropomyosin. Results The optimal extraction process of Sargassum carpophyllum polyphenols was as follows: The extraction temperature was 40℃ and the extraction time was 10 h. With 75% ethanol at 1:16 (m/V), the extraction yield of Sargassum carpophyllum polyphenols reached 18.18 mg/g. After purification, Sargassum carpophyllum polyphenols with a purity of 71.93% were obtained, which contained salicylic acid monohydrate, 3,4-dihydroxy-5-sulfoxybenzoic acid and gallic acid. Sargassum carpophyllum polyphenols could inhibit the sensitization of tropomyosin by binding aromatic amino acids and fluorescent amino acids in tropomyosin, blocking and destroying epitopes of tropomyosin. Conclusion In this study, the extraction and purification process of Sargassum carpophyllum polyphenols is optimized, and the obtained Sargassum carpophyllum polyphenols can significantly reduce the allergenicity of tropomyosin and show strong anti-allergic activity.