细胞壁缺失对副溶血性弧菌生物学特性的影响
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国家重点研发计划课题“生食水产品加工储运过程中生物性危害防控技术与示范”(2019YFD0901702)


Effects of Cell Wall Deletion on the Biological Characteristics of Vibrio parahaemolyticus
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    摘要:

    副溶血性弧菌是沿海地区重要的食源性致病菌之一,该菌的抗药性问题逐渐凸显,急需开发安全、有效、具有新作用位点的抑菌剂。本文以细胞壁为潜在靶点,采用肉桂醇作为抑菌剂研究细胞壁缺失对副溶血性弧菌生物学特性的影响。通过测定去壁菌体在最小抑菌浓度、生长曲线、细胞膜完整性、内膜通透性和外膜蛋白竞争试验中的变化,探寻肉桂醇作用于菌体的部位和抑制方式,并通过药敏试验验证获得的去壁细胞。结果表明:细胞壁缺失使肉桂醇对副溶血性弧菌的MIC降低为未处理组的1/8。肉桂醇对不同状态的副溶血性弧菌均有很强的抑制作用,可造成细胞膜损伤,引起胞内特征大分子外漏。硫酸粘杆菌素和去壁处理能够使肉桂醇失去作用靶点。外膜蛋白竞争试验进一步证实,肉桂醇能与外膜蛋白产生相互作用。去壁处理使原本作用于细胞壁的抗生素失去药效,表明去壁操作无误。结论:肉桂醇可以作用于副溶血性弧菌的细胞膜和外膜,可能与外膜蛋白发生靶向结合。外膜是小分子进入阴性菌的第1道屏障,可作为抑菌剂开发的新靶点。

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    Vibrio parahaemolyticus is one of the important food-borne pathogens in coastal areas, and the problem of drug resistance of this bacterium is becoming more and more serious, it is urgent to develop safe, effective bacteriostatic agents with new action sites. In this paper, the effect of cell wall deletion on the biological characteristics of V. parahaemolyticus was studied with the cell wall as a potential target and cinnamyl alcohol as a bacteriostatic agent. The position and mode of the inhibition of cinnamyl alcohol on the bacteria were explored by measuring the changes in the minimal inhibitory concentration, growth curve, cell membrane integrity, inner membrane permeability and outer membrane protein competition test, and obtained dewalled cells were verified by drug sensitivity tests. The results showed that the cell wall deletion reduced the MIC of cinnamyl alcohol to V. parahaemolyticus to 1/8 of the untreated group. Cinnamyl alcohol has a strong inhibitory effect on V. parahaemolyticus in different states, which could cause cell membrane damage and cause intracellular characteristic macromolecular leakage. Colistin sulfate and wall removal could make cinnamyl alcohol lose its target. The outer membrane protein competition test further confirmed that cinnamyl alcohol could interact with outer membrane proteins. The de-walling treatment made the antibiotics that had been acting on the cell wall ineffective, indicating that the de-walling operation was done correctly. Conclusion: Cinnamyl alcohol can act on the cell membrane and outer membrane of V. parahaemolyticus, which may be targeted to the outer membrane protein. The outer membrane is the first barrier for small molecules to enter the Gram-negative bacteria and can be used as a new target for the development of bacteriostatic agents.

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刘雪飞;吴思楠;李秀霞;谢晶;牟伟丽;郭晓华;李学鹏;励建荣.细胞壁缺失对副溶血性弧菌生物学特性的影响[J].中国食品学报,2021,21(5):23-30

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  • 在线发布日期: 2021-06-07
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