文献类型：期刊文献

中文题名：凡纳滨对虾多酚氧化酶的纯化鉴定与生物信息学分析

英文题名：Purification and Identification of PPO from Litopenaeus vannamei and Its Bioinformatics Analysis

作者：魏帅[1,2,3];刘媛[1,2,3];刘蒙娜[1,2,3];孙钦秀[1,2,3];刘书成[1,2,3];吉宏武[1,2,3];郝记明[1,2,3];邓楚津[1,2,3];毛伟杰[1,2,3]

机构：[1]广东海洋大学食品科技学院,广东省水产品加工与安全重点实验室,广东省海洋食品工程技术研发中心,广东省海洋生物制品工程重点实验室,水产品深加工广东普通高校重点实验室,广东湛江524088;[2]海洋食品精深加工关键技术省部共建协同创新中心,大连工业大学,辽宁大连116034;[3]南方海洋科学与工程广东省实验室(湛江),广东湛江524088

年份：2020

卷号：40

期号：4

起止页码：100

中文期刊名：广东海洋大学学报

外文期刊名：Journal of Guangdong Ocean University

收录：CSTPCD

基金：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(2019KCXTD011)。

语种：中文

中文关键词：凡纳滨对虾;多酚氧化酶;质谱;鉴定;生物信息学

外文关键词：Litopenaeus vannamei;PPO;mass spectrometry;identification;bioinformatics

中文摘要：【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。

外文摘要：【Objective】To isolate and purify the PPO from Litopenaeus vannamei,the biological information was identified and analyzed by using mass spectrometry.【Methods】PPO was isolated and purified from the shrimp cephalothorax by using ammonium sulfate precipitation and column chromatography.The molecular weight was determined by electrophoresis,and the amino acid sequence was determined by liquid chromatography-electrospray ionization-tandem mass spectrometry(LC-ESIMS/MS).Based on these results,homology modeling was constructed and the evaluations were performed.【Results and Conclusion】Purified PPO was obtained with a purification of 65.7 folds,the specific activity of PPO was 650.1 U/mg protein and with a yield of 29.5%.The molecular weight of PPO was about 72 ku.The similarity of amino acid sequence between the purified protein and Propolyphenoloxidase of L.vannamei was 65%with a matching score of 11321 points by identification and analysis of mass spectrometry and basic local alignment search tool(blast)sequence comparison.The results have confirmed that the purified protein was PPO.PPO is an amphoteric protein and it has a higher hydrophilicity than hydrophobicity.The secondary structure of PPO mainly consists ofα-helix and random coil.In conclusion,the predicted three-dimensional structure of PPO had a high reliability.