Shen Lixin, Duan Kangmin
Wei Sheng Wu Xue Bao. 2014 Dec 4;54(12):1419-28.
To investigate the function of gene PA2580 in Pseudomonas aeruginosa PAO1.
We constructed a PA2580 knockout mutant of PAO1 and a complemented strain of the mutant. We studied the function of the gene PA2580 using both genetic and biochemical methods, including antibiotic minimum inhibition concentration (MIC) comparison, measurement of gene expression levels in different conditions, protein expression and purification in vitro and enzymatic activity detection.
PA2580 mutant was more sensitive to carbenicillin, chloramphenicol and ciprofloxacin. PA2580 expression was regulated by sub-inhibitory concentrations of antibiotics. PA2580 protein reduced various quinones efficiently using NADPH as the electron donor. PA2580 mutant was more sensitive to hydrogen peroxide and the mutant showed decreased expression of catalase. These results indicate that PA2580 is involved in the tolerance of oxidative stress in P. aeruginosa.
The P. aeruginosa PA2580 protein physiologically functions as an NADPH quinone reductase which plays an important role in dealing with environment stress.
研究基因PA2580在铜绿假单胞菌PAO1中的功能。
构建PAO1的PA2580基因敲除突变体及其互补菌株。采用遗传学和生物化学方法研究PA2580基因的功能，包括抗生素最低抑菌浓度（MIC）比较、不同条件下基因表达水平测定、体外蛋白表达与纯化以及酶活性检测。
PA2580突变体对羧苄青霉素、氯霉素和环丙沙星更敏感。PA2580的表达受亚抑菌浓度抗生素的调控。PA2580蛋白以NADPH作为电子供体，能有效还原多种醌类物质。PA2580突变体对过氧化氢更敏感，且该突变体中过氧化氢酶的表达降低。这些结果表明PA2580参与了铜绿假单胞菌对氧化应激的耐受性。
铜绿假单胞菌PA2580蛋白在生理上作为一种NADPH醌还原酶，在应对环境应激中发挥重要作用。
