Master Program of Microbiology and Immunology, School of Medicine, Tzu Chi University, Hualien 97004, Taiwan.
Department of Microbiology, School of Medicine, Tzu Chi University, Hualien 97004, Taiwan.
Int J Mol Sci. 2021 Sep 14;22(18):9921. doi: 10.3390/ijms22189921.
Most bacteria possess alcohol dehydrogenase (ADH) genes ( genes) to mitigate alcohol toxicity, but these genes have functions beyond alcohol degradation. Previous research has shown that ADH can modulate quorum sensing in , a rising opportunistic pathogen. However, the number and nature of genes in have not yet been fully characterized. We identified seven alcohol dehydrogenases (NAD-ADHs) from ATCC 19606, and examined the roles of three iron-containing ADHs, ADH3, ADH4, and ADH6. Marker-less mutation was used to generate , , and single, double, and triple mutants. Disrupted mutants failed to grow in ethanol-, 1-butanol-, or 1-propanol-containing mediums, and recombinant ADH4 exhibited strongest activity against ethanol. Stress resistance assays with inorganic and organic hydroperoxides showed that and were key to oxidative stress resistance. Virulence assays performed on the model organism revealed that mutants had comparable virulence to wild-type, while and mutants had reduced virulence. The results suggest that ADH4 is primarily involved in alcohol metabolism, while ADH3 and ADH6 are key to stress resistance and virulence. Further investigation into the roles of other ADHs in is warranted.
大多数细菌都拥有能够减轻酒精毒性的醇脱氢酶（ADH）基因（genes），但这些基因的功能远不止于酒精降解。先前的研究表明，ADH 可以调节渐增的机会性病原体的群体感应。然而，尚未完全确定 中的 ADH 基因的数量和性质。我们从 ATCC 19606 中鉴定出了 7 种醇脱氢酶（NAD-ADHs），并研究了三种含铁的 ADH（ADH3、ADH4 和 ADH6）的作用。使用无标记突变生成了 、 、和 单、双和三突变体。破坏 突变体无法在含有乙醇、1-丁醇或 1-丙醇的培养基中生长，重组 ADH4 对乙醇表现出最强的活性。用无机和有机过氧化物进行的应激抗性测定表明， 和 对于氧化应激抗性至关重要。在 模式生物上进行的毒力测定表明， 突变体与野生型具有相当的毒力，而 和 突变体的毒力降低。结果表明，ADH4 主要参与酒精代谢，而 ADH3 和 ADH6 是应激抗性和毒力的关键。需要进一步研究其他 ADH 在 中的作用。
