波罗的海希瓦氏菌和杀鲑气单胞菌致腐性及TorA还原酶差异比较
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S984.1

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浙江省自然科学基金(Y19C200005);浙江省教育厅项目(Y201839893);浙江省分析测试项目(LGC19C200004)


Comparison of spoilage potential and TorA protein betweenShewanella baltica and Aeromonas salmonicida
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    摘要:

    为了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,本实验比较分析了大黄鱼源波罗的海希瓦氏菌和杀鲑气单胞菌在28℃和4℃下的生长及三甲胺(TMA)、生物胺和挥发性盐基氮(TVB-N)的生成;通过PCR技术扩增2种腐败菌的氧化三甲胺还原酶基因(torA),利用生物信息学比较TorA蛋白的相似性、理化特性和蛋白空间结构。结果显示,杀鲑气单胞菌在28℃生长较快,而波罗的海希瓦氏菌在4℃生长更快。相对于杀鲑气单胞菌形成较高的尸胺,波罗的海希瓦氏菌产生更多TMA和腐胺,在冷藏鱼汁中积累更高TVB-N。同时在波罗的海希瓦氏菌和杀鲑气单胞菌中分别扩增出2 490和1 959 bp的torA基因,2种TorA蛋白与同属菌相似性高于97%,而二者相似性仅为36.90%。波罗的海希瓦氏菌TorA蛋白的分子量和等电点分别为92.3 ku和6.52,甘氨酸含量最高,而杀鲑气单胞菌中TorA蛋白的分子量和等电点分别为90.6 ku和6.74,丙氨酸含量最高,蛋白结构差异明显。且希瓦氏菌中torA 和鸟氨酸脱羧酶(DOC)基因表达量分别为气单胞菌的1.26和19.04倍。可见,波罗的海希瓦氏菌和杀鲑气单胞菌为海产品嗜冷腐败菌,其中希瓦氏菌胺类代谢能力更强,与其TorA特定理化特性和高表达量相关。本研究为揭示海产品微生物的致腐机制提供理论支持。

    Abstract:

    To evaluate the differences of spoilage potential of Shewanella and Aeromonas in refrigerated seafood, growth curves and production of trimethylamine (TMA), biogenic amines and total volatile basic nitrogen (TVB-N) in S. baltica and A. salmonicida isolated from Larimichthys crocea were measured at 28℃ and 4℃. And trimethylamine reductase (torA) gene in the two strains were amplified, and similarity, physicochemical properties and spatial structures of TorA protein were compared by bioinformatics. A. salmonicida grew faster at 28℃, while S. baltica grew faster at 4℃. Compared with more amount of cadaverine in A. salmonicida, S. baltica produced the higher levels of TMA, biogenic amines, and TVB-N in refrigerated fish juice. The torA genes of 2 490 bp and 1 959 bp were amplified from S. baltica and A. salmonicida respectively, and similarity of TorA protein in its same genus was higher than 97%, and similarity between the two strains was only 36.90%. The molecular weight and isoelectric point of TorA protein in S. baltica were about 92.3 ku and 6.52, while those were about 90.6 ku and 6.74 in A. salmonicida. The highest amino acid of the predicted protein was glycine and alanine in two strains respectively, resulting in the significant difference of protein spatial structure. The expression levels of torA and ornithine decarboxylase ODC genes in S. baltica were 1.26 and 19.04 times compared to A. salmonicida. Thus, S. baltica and A. salmonicida are psychrophilic spoilage strains in marine products, in which S. baltica has stronger ability to produce amine metabolites due to specific physicochemical properties of TorA protein and its high expression. The study provides theoretical support for revealing the mechanism of microbial spoilage in seafood.

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毕伟伟,冯立芳,石双妮,缪晨豪,朱军莉,田迪英.波罗的海希瓦氏菌和杀鲑气单胞菌致腐性及TorA还原酶差异比较[J].水产学报,2020,44(3):495~504

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  • 收稿日期:2019-01-24
  • 最后修改日期:2019-08-01
  • 录用日期:2019-08-10
  • 在线发布日期: 2020-03-10
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