Kouzuma Atsushi, Endoh Takayuki, Omori Toshio, Nojiri Hideaki, Yamane Hisakazu, Habe Hiroshi
Biotechnology Research Center, The University of Tokyo, Tokyo, Japan.
FEMS Microbiol Lett. 2007 Oct;275(1):175-81. doi: 10.1111/j.1574-6968.2007.00882.x. Epub 2007 Aug 15.
Many bacteria living in soil have developed the ability to use a wide variety of organosulfur compounds. Pseudomonas putida strain DS1 is able to utilize dimethyl sulfide as a sulfur source via a series of oxidation reactions that sequentially produce dimethyl sulfoxide, dimethyl sulfone (DMSO2), methanesulfonate, and sulfite. To isolate novel genes involved in DMSO2 utilization, a transposon-based mutagenesis of DS1 was performed. Of c. 10,000 strains containing mini-Tn5 inserts, 11 mutants lacked the ability to utilize DMSO2, and their insertion sites were determined. In addition to the cysNC, cysH, and cysM genes involved in sulfate assimilation, the ptsP gene encoding the phosphoenolpyruvate:sugar phosphotransferase system (PTS) family protein EI(Ntr) was identified, which is necessary for DMSO2 utilization. Using quantitative reverse transcriptase-polymerase chain reaction analysis, it was demonstrated that the expression of the sfn genes, necessary for DMSO2 utilization, was impaired in the ptsP disruptant. To the authors' knowledge, this is the first report of a PTS protein that is involved in bacterial assimilation of organosulfur compounds.
许多生活在土壤中的细菌已经发展出利用多种有机硫化合物的能力。恶臭假单胞菌DS1菌株能够通过一系列氧化反应将二甲基硫醚用作硫源,这些反应依次产生二甲基亚砜、二甲基砜(DMSO2)、甲磺酸盐和亚硫酸盐。为了分离参与DMSO2利用的新基因,对DS1进行了基于转座子的诱变。在约10,000个含有mini-Tn5插入片段的菌株中,有11个突变体缺乏利用DMSO2的能力,并确定了它们的插入位点。除了参与硫酸盐同化的cysNC、cysH和cysM基因外,还鉴定出编码磷酸烯醇丙酮酸:糖磷酸转移酶系统(PTS)家族蛋白EI(Ntr)的ptsP基因,该基因是DMSO2利用所必需的。通过定量逆转录-聚合酶链反应分析表明,DMSO2利用所必需的sfn基因的表达在ptsP缺失突变体中受损。据作者所知,这是关于一种参与细菌有机硫化合物同化的PTS蛋白的首次报道。