Department of Applied and Environmental Microbiology, Ludong University, Yantai, 264000, China.
Affiliated Hospital, Ludong University, Yantai, 264025, China.
Arch Microbiol. 2020 Aug;202(6):1507-1515. doi: 10.1007/s00203-020-01837-8. Epub 2020 Mar 28.
Pyocyanin produced by Pseudomonas aeruginosa is a key virulence factor that often causes heavy damages to airway and lung in patients. Conversion of phenazine-1-carboxylic acid to pyocyanin involves an extrametabolic pathway that contains two enzymes encoded, respectively, by phzM and phzS. In this study, with construction of the rpoS-deficient mutant, we first found that although phenazine production increased, pyocyanin produced in the mutant YTΔrpoS was fourfold much higher than that in the wild-type strain YT. To investigate this issue, we constructed phzM-lacZ fusion on a vector and on the chromosome. By quantifying β-galactosidase activities, we confirmed that expression of the phzM was up-regulated when the rpoS gene was inactivated. However, no changes occurred in the expression of phzS and phzH when the rpoS was knocked out. Taken together, overproduction of the SAM-dependent methyltransferase (PhzM) might contribute to the increased pyocyanin in the absence of RpoS in P. aeruginosa.
铜绿假单胞菌产生的绿脓菌素是一种关键的毒力因子,常导致气道和肺部严重损伤。吩嗪-1-羧酸向绿脓菌素的转化涉及一条细胞外代谢途径,该途径分别由 phzM 和 phzS 编码的两种酶所包含。在这项研究中,通过构建 rpoS 缺陷突变体,我们首先发现,尽管吩嗪产量增加,但突变体 YTΔrpoS 产生的绿脓菌素比野生型菌株 YT 高出四倍。为了研究这个问题,我们在载体和染色体上构建了 phzM-lacZ 融合。通过定量β-半乳糖苷酶活性,我们证实当 rpoS 基因失活时,phzM 的表达被上调。然而,当 rpoS 被敲除时,phzS 和 phzH 的表达没有变化。总之,SAM 依赖性甲基转移酶(PhzM)的过度表达可能导致铜绿假单胞菌中缺乏 RpoS 时绿脓菌素的增加。