文章摘要
波罗的海希瓦氏菌和杀鲑气单胞菌致腐性及TorA差异比较
Comparison of Spoilage Potential and TorA Protein between Shewanella baltica and Aeromonas salmonicida
投稿时间:2019-01-24  修订日期:2019-08-01
DOI:
中文关键词: 希瓦氏菌  气单胞菌  致腐性  TorA
英文关键词: Shewanella  Aeromonas  spoilage potential  TorA
基金项目:省协同创新中心项目(2017SICR105);浙江省教育厅项目(Y201839893),浙江省分析测试项目(LGC19C200004)。*通讯作者。Tel:+86-571-88071024-8588;E-mail:junlizhu0305@163.com
作者单位E-mail
毕伟伟 浙江工商大学食品与生物工程学院 1032984616@qq.com 
石双妮 浙江工商大学食品与生物工程学院  
繆晨豪 浙江工商大学食品与生物工程学院  
朱军莉 浙江工商大学食品与生物工程学院 junlizhu0305@163.com 
田迪英 浙江工商大学食品与生物工程学院  
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中文摘要:
      [目的]为研究冷藏海产品中腐败菌希瓦氏菌和气单胞菌的致腐性差异。[方法]测定大黄鱼源波罗的海希瓦氏菌(Shewanella baltica)和杀鲑气单胞菌(Aeromonas salmonicida)在28℃和4℃下生长曲线,三甲胺(TMA)、生物胺的生成及挥发性盐基氮(TVB-N)的积累。PCR扩增两种腐败菌的氧化三甲胺(TMAO)还原酶基因(torA),生物信息学比较TorA蛋白的相似性、理化特性和蛋白空间结构。[结果]结果表明,在28℃培养下A. salmonicida生长较快,而在4℃培养下S. baltica生长更快。相对于A. salmonicida,S. baltica产生较高的TMA和生物胺,在冷藏鱼汁中积累更高的TVB-N,表明S. baltica具有更强的胺类生成能力。同时在S. baltica和A. salmonicida分别扩增出2490bp和1959bp的torA基因。生物信息学分析显示,两种TorA蛋白与同属菌相似性高于97%,而两者间的相似性仅为36.90%,S. baltica中TorA蛋白的分子量和等电点约为92.3k和6.52,甘氨酸含量最高,而气单胞菌中约为90.6k和6.74,丙氨酸含量最高,两个TorA蛋白的空间结构差异明显,但均为分泌蛋白,表达量相似。[结论] S. baltica和A. salmonicida为海产品嗜冷腐败菌,其中前者形成更强的胺类代谢物,并且两者TorA蛋白的理化特性和空间结构差异,使TMAO三甲胺酶表现不同活性。研究为揭示海产品嗜冷腐败菌的致腐机制提供理论支持。
英文摘要:
      [Objective] To evaluate the differences of spoilage potential of Shewanella and Aeromonas in refrigerated seafood. [Methods] Growth curves, the formation of trimethylamine (TMA), biogenic amines and the accumulation of volatile base nitrogen (TVB-N) of Shewanella baltica and Aeromonas salmonicida isolated from Larimichthys crocea were measured at 28 °C and 4 °C. And trimethylamine reductase (torA) gene in two spoilage bacteria was amplified, and similarity, physicochemical properties and protein spatial structure of TorA protein were compared by bioinformatics. [Results] The results showed that A. salmonicida grew faster at 28 °C, while S. baltica grew faster at 4 °C. Compared with A. salmonicida, S.baltica produced the higher levels of TMA and biogenic amines, and accumulates higher TVB-N content in sterile fish juice, which indicated that S.baltica had stronger ability to produce amine metabolites. The torA genes of 2490 bp and 1959 bp were amplified from S.baltica and A. salmonicida respectively. Bioinformatics analysis showed that the similarity between TorA proteins in the isolates and same genus was higher than 97%, and between the two isolates were only 36.90%. The molecular weight and isoelectric point of TorA protein in S.baltica were about 92.3k and 6.52, with highest content of glycine, while those of A. salmonicida were about 90.6k and 6.74, with alanine as the highest amino acid. The spatial structure of TorA protein in two isolates was significantly different, however, they were both secreting proteins with similar expression levels. [Conclusion] S.baltica and A. salmonicida are psychrophilic spoilage bacteria in marine products, and S.baltica has stronger production of amine metabolites. TorA protein in two isolates exhibited the different similarity and physicochemical properties, resulting in different activity of TMAO trimethylamine. The study provides theoretical support for revealing the spoilage mechanism involving in psychrophilic spoilage bacteria.
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