酸胁迫条件对粪肠球菌Gr17代谢合成细菌素的影响
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(北京工商大学 北京食品营养与人类健康高精尖创新中心 北京市食品添加剂工程技术研究中心 北京 100048)

作者简介:

刘国荣(1983—),女,博士,教授 E-mail: liuguorong@th.btbu.edu.cn

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基金项目:

国家重点研发计划项目(2019YFC1606200);国家自然科学基金面上项目(31871772);北京市自然科学基金面上项目(6192003)


Effects of Acid Stress Conditions on Bacteriocin Synthesis by Metabolism of Enterococcus faecalis Gr17
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(Beijing Advanced Innovation Center for Food Nutrition and Human Health, Beijing Engineering and Technology Research Center of Food Additives, Beijing Technology and Business University, Beijing 100048)

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    摘要:

    目的:粪肠球菌Gr17分离自中国传统发酵酸鱼,可以代谢合成新型广谱IIa类细菌素enterocin Gr17,有作为发酵食品功能性菌株的巨大应用潜力,然而该菌株应用于实际发酵环境时会面临各种胁迫条件,酸胁迫是其中主要的胁迫因素之一。本研究旨在探讨不同酸胁迫条件对菌株Gr17生长及代谢合成细菌素的影响。方法:在筛选可正向影响细菌素合成的酸胁迫条件参数基础上,测定该条件下细菌素生物合成相关基因的表达水平变化,分析其影响机制。结果:与正常生长条件pH 7.0相比,在pH 4.5~6.5的酸胁迫条件下,菌株Gr17的菌体密度会随pH值的下降而降低;pH 5.0~6.5的酸胁迫条件可明显正向提高细菌素的分泌合成量,其中pH 5.5的酸胁迫条件最为明显。在此条件下,细菌素生物合成相关ABC转运系统as-48H、as-48G、as-48F、as-48E基因表达在处理时间48 h内没有显著变化,种内群体感应调控系统自诱导肽基因entIP在8 h时表达量明显提高,双组分基因entPK、entR以及细菌素编码基因entGr17在12 h时表达量明显提高。结论:基于以上结果,推测pH 5.5的酸胁迫条件可以促进自诱导肽的分泌表达,正向调控种内群体感应系统,从而增加细菌素的合成。本研究结果对于从分子水平上全面揭示实际发酵环境中乳酸菌细菌素的合成调控行为,最大限度地提高产细菌素乳酸菌在发酵生产中的应用价值具有重要的科学和现实意义。

    Abstract:

    Objective: Enterococcus faecalis Gr17 is isolated from Chinese traditional fermented sour fish. It can be metabolized into a new broad-spectrum IIa bacteriocin enterocin Gr17, which has great application potential as a functional strain of fermented food. However, when the strain is used in the actual fermentation environment, it will inevitably face various stress conditions. Acid stress is one of the main stress factors. The purpose of this research was to investigate the effects of different acid stress conditions on the growth and bacteriocin biosynthesis of Gr17. Methods: On the basis of screening the acid stress parameters which could positively affect bacteriocin biosynthesis, the expression level of genes related to bacteriocin biosynthesis under the condition was determined and the influence mechanism was preliminarily analyzed. Results: The results showed that the cell density of Gr17 decreased with the decrease of pH value at the condition of pH 4.5~6.5 compared with the normal growth condition of pH 7.0. Under the condition of pH 5.0~6.5, the secretion and synthesis of bacteriocin was increased. Acid stress at pH 5.5 could significantly increase the secretion and synthesis of bacteriocin. Under these conditions, there was no significant change in the gene expression of ABC transport system associated with bacteriocin biosynthesis in as-48E, as-48F, as-48G and as-48H within 48 h of treatment. The expression of self-induced peptide gene entIP in quorum sensing regulation system increased significantly at 8 h. The expression of two-component genes entPK, entR and bacteriocin coding gene entGr17 increased obviously at 12 h. Conclusion: Based on the above results, it is speculated that acid stress at pH 5.5 could promote the secretion and expression of self-induced peptides, positively regulate quorum sensing system, thus increasing the synthesis of bacteriocin. The results of this study have important scientific and practical significance for revealing the regulation behavior of bacteriocin synthesis in the actual fermentation environment at the molecular level, and maximizing the application value of bacteriocin producing lactic acid bacteria in fermentation production.

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刘国荣,聂蓉,段娇娇,王成涛.酸胁迫条件对粪肠球菌Gr17代谢合成细菌素的影响[J].中国食品学报,2022,22(5):18-26

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  • 收稿日期:2021-05-04
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  • 在线发布日期: 2022-06-15
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