Mishra Mukti Nath, Thirunavukkarasu Nagarajan, Sharma Indra Mani, Jagnnadham Medicharla V, Tripathi Anil K
School of Biotechnology, Faculty of Science, Banaras Hindu University, Varanasi, India.
FEMS Microbiol Lett. 2008 Oct;287(2):221-9. doi: 10.1111/j.1574-6968.2008.01325.x. Epub 2008 Aug 27.
Azospirillum brasilense Sp7 has been shown to overproduce carotenoids if the anti-sigma factor (anti-sigma(E))-encoding gene is inactivated. The anti-sigma mutant (Car-1) of A. brasilense Sp7 was more tolerant to the stresses generated by elevated temperature (40 degrees C), PEG-200 (30 mg mL(-1)) and the antibacterial agent Polymyxin-B (PMB, 25 microg mL(-1)) but not to elevated salinity (15 mg mL(-1)). Inhibition of carotenoid synthesis by diphenylamine inhibited the ability of the mutant to tolerate all the three stresses. Out of the four stress agents, only elevated temperature and salinity induced the rpoE promoter and increased the carotenoid content in Sp7 as well as in the Car-1 mutant. Comparison of the membrane permeability of the parent and the mutant by a PMB-N-phenyl-1-naphthylamine coupled assay showed that the presence of carotenoids in the mutant reduced the permeability of their membranes. Our study indicates that the carotenoid synthesis, which is under the control of extracytoplasmic function sigma factor (sigma(E)) in A. brasilense Sp7, plays a positive role in tolerating elevated temperature, the antibacterial peptide and PEG-200.
如果编码抗σ因子(抗σ(E))的基因失活,巴西固氮螺菌Sp7已被证明会过量产生类胡萝卜素。巴西固氮螺菌Sp7的抗σ突变体(Car-1)对高温(40℃)、PEG-200(30 mg mL(-1))和抗菌剂多粘菌素B(PMB,25 μg mL(-1))产生的胁迫更具耐受性,但对盐度升高(15 mg mL(-1))不具耐受性。二苯胺对类胡萝卜素合成的抑制作用抑制了突变体耐受所有三种胁迫的能力。在这四种胁迫因子中,只有高温和盐度诱导了rpoE启动子,并增加了Sp7以及Car-1突变体中的类胡萝卜素含量。通过PMB-N-苯基-1-萘胺偶联测定法比较亲本和突变体的膜通透性,结果表明突变体中类胡萝卜素的存在降低了其膜的通透性。我们的研究表明,巴西固氮螺菌Sp7中由胞外功能σ因子(σ(E))控制的类胡萝卜素合成在耐受高温、抗菌肽和PEG-200方面发挥了积极作用。