Department of Food and Nutritional Sciences, The University of Reading, PO Box 226, Whiteknights, Reading, RG6 6AP, UK.
Bacterial Stress Response Group, Microbiology, School of Natural Sciences, National University of Ireland, Galway, Galway, Ireland.
Food Microbiol. 2020 Sep;90:103481. doi: 10.1016/j.fm.2020.103481. Epub 2020 Mar 2.
The Glutamate Decarboxylase (GAD) system is important for survival of L. monocytogenes and other microorganisms under acidic conditions. Environmental conditions influence the function of the GAD system. Until now, the only conditions known to lead to increased transcription of the GAD system are the stationary phase in rich media and anoxic conditions. Previously, we showed that transcription of the GAD system requires unidentified compounds other than glutamate present in rich media. Following a test looking at various compounds we identified for first time that peptone, tryptone and casamino acids activate the GAD system under oxic conditions suggesting that amino acid(s) other than glutamate and/or peptides are important for the above process. The defined medium, where the GAD system is inactive, once it is supplemented with the above compounds results in an active intracellular and extracellular GAD system and increased acid resistance. Through functional genomics we show that these compounds are required for GadD2 activity and although we previously showed that GadD3 is active part of the intracellular GAD system, the supplementation did not activate this gene. The above is explained by the fact that only gadD2 transcription was upregulated by these compounds while the transcription of gadD1 and gadD3 remained unaffected. Together our results show that the L. monocytogenes GadD2 decarboxylase is activated in the presence of amino acids or peptides other than glutamate, a finding that has important implications for acid tolerance and food safety.
谷氨酸脱羧酶(GAD)系统对于李斯特菌和其他微生物在酸性条件下的生存至关重要。环境条件会影响 GAD 系统的功能。到目前为止,已知唯一能导致 GAD 系统转录增加的条件是丰富培养基中的静止期和缺氧条件。此前,我们已经证明,GAD 系统的转录需要丰富培养基中除谷氨酸以外的未知化合物。在对各种化合物进行测试后,我们首次发现蛋白胨、胰蛋白胨和氨基酸肉汤能在有氧条件下激活 GAD 系统,这表明除谷氨酸和/或肽以外的氨基酸对于上述过程很重要。在补充上述化合物之前,定义培养基中的 GAD 系统是不活跃的,这会导致细胞内和细胞外 GAD 系统活跃,并增强耐酸性。通过功能基因组学,我们表明这些化合物是 GadD2 活性所必需的,尽管我们之前已经表明 GadD3 是细胞内 GAD 系统的活性部分,但补充这些化合物并没有激活该基因。这是因为这些化合物仅上调了 gadD2 的转录,而 gadD1 和 gadD3 的转录不受影响。综上所述,我们的研究结果表明,除了谷氨酸之外,其他氨基酸或肽也能激活李斯特菌的 GadD2 脱羧酶,这一发现对于耐酸性和食品安全具有重要意义。