牛肉低温贮藏环境中沙门氏菌诱导耐酸响应的存在程度及其产生机制

摘要/Abstract

摘要： 冷鲜牛肉生产过程中弱酸的压力环境能够引发沙门氏菌产生诱导耐酸响应（Acid tolerance response, ATR），危害肉品安全。但该现象一旦产生，其在牛肉低温成熟与贮藏过程中的消长规律仍不明确、调控机制尚不清楚。为了探究牛肉生产和销售过程中沙门氏菌ATR的存在程度及其产生机制，本实验探究了微酸诱导、双组分基因敲除、低温长期贮藏对沙门氏菌耐酸能力的影响，同时借助λRed同源重组、RT-qPCR及氨基酸添加实验探索菌株耐酸性产生的内在机理。结果表明微酸诱导能够显著增强沙门氏菌的耐酸能力（P < 0.05），并且ATR一旦形成，在牛肉低温贮藏（4 ℃）的过程中至少可以维持1周，对食品安全有极大危害。牛肉培养基作为微酸的细菌生长介质，在低温下其本身并不能引发沙门氏菌产生诱导耐酸反应，说明低温处理可能是抑制沙门氏菌ATR的重要方法。RT-qPCR和氨基酸添加实验表明，沙门氏菌的双组分系统PhoP/PhoQ和PmrA/PmrB均参与酸性环境的感知，并能通过调控精氨酸脱羧和赖氨酸脱羧系统来提高菌株的耐酸性，这从氨基酸代谢角度解释了沙门氏菌ATR的产生机制，同时也揭示了食品基质提升微生物耐酸性这一表观现象的内在机理。

关键词: 牛肉, 沙门氏菌, 诱导耐酸响应, 双组分调控系统, PhoP/PhoQ, PmrA/PmrB, 食品安全

Abstract: Mild acidic and nutrient environment of chilled beef may induce acid tolerance response (ATR) of Salmonella, posing a great risk to meat quality and safety. But the existence and mechanism of this phenomenon during the aging and storage of beef was still unclear. The influence of acid induction, two-component systems and the low-temperature storage on the formation of ATR were evaluated, λRed recombination system, RT-qPCR and amino acid addition experiments were also conducted to explore the mechanism. The results showed that mild acid induction significantly enhanced the acid tolerance of Salmonella (P < 0.05), and once induced, it could be maintained for at least 1 week at 4 ℃. As a mild acidic incubation medium, beef does not induce the ATR at 4 ℃, indicating that low-temperature treatment may be an important method to inhibit ATR in Salmonella typhimurium. RT-qPCR and amino acid addition experiments showed that PhoP/PhoQ and PmrA/PmrB are involved in the perception of the acidic environment, improving the bacterial acid resistance by regulating arginine decarboxylation and lysine decarboxylation systems. This explains the formation of the ATR from the aspect of amino acid metabolism, which may also reveal an explanation of why substrates enhance the acid resistance of some bacteria in food.

Key words: Beef, Salmonella, acid tolerance response, two-component regulatory system, PhoP/PhoQ, PmrA/PmrB, food safety

中图分类号:

TS251.5+2

郎晨晓张一敏朱立贤梁荣蓉毛衍伟杨啸吟韩广星罗欣董鹏程. 牛肉低温贮藏环境中沙门氏菌诱导耐酸响应的存在程度及其产生机制[J]. 食品科学, 0, (): 0-0.

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