凡纳滨对虾多酚氧化酶的纯化鉴定与生物信息学分析Purification and Identification of PPO from Litopenaeus vannamei and Its Bioinformatics Analysis
魏帅,刘媛,刘蒙娜,孙钦秀,刘书成,吉宏武,郝记明,邓楚津,毛伟杰
摘要(Abstract):
【目的】从凡纳滨对虾(Litopenaeus vannamei)中分离纯化多酚氧化酶(Polyphenol Oxidase,PPO),进行质谱鉴定,分析其生物学信息。【方法】采用硫酸铵沉淀结合柱层析法从虾头胸部中提取纯化PPO,电泳法确定其分子质量,采用液相质谱联用法分析氨基酸序列结构,根据分析结果,同源建模并进行评估。【结果与结论】获得了高纯度的PPO,其纯化倍数为65.7,酶比活力达650.1 U/mg蛋白,得率为29.5%。PPO分子质量约为72 ku。经质谱检测分析和basic local alignment search tool(BLAST)序列比对,纯化蛋白与凡纳滨对虾酚氧化酶原的氨基酸序列相似性为65%,匹配度分值为11 321,确认为PPO。PPO是一种亲水性大于疏水性的两性蛋白,二级结构以α-螺旋和无规则卷曲为主,建立的PPO三维结构模型可靠性较高。
关键词(KeyWords): 凡纳滨对虾;多酚氧化酶;质谱;鉴定;生物信息学
基金项目(Foundation): 广东省区域联合基金-青年基金项目(2019A1515110419);; 国家重点研发计划资助(2019YFD0902003);; 国家自然科学基金面上项目(31771997);; 南方海洋科学与工程广东省实验室(湛江)资助(ZJW-2019-06);; 广东海洋大学创新强校重大培育项目(GDOU2017052603);; 国家虾蟹产业技术体系(G-48);; 广东省普通高校青年创新人才项目(2019KQNCX040);; 广东普通高等学校海洋食品绿色加工技术研究团队(2019KCXTD 011)
作者(Author): 魏帅,刘媛,刘蒙娜,孙钦秀,刘书成,吉宏武,郝记明,邓楚津,毛伟杰
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