De Spiegeleer P, Sermon J, Lietaert A, Aertsen A, Michiels C W
Laboratory of Food Microbiology, Katholieke Universiteit Leuven, Leuven, Belgium.
J Appl Microbiol. 2004;97(1):124-33. doi: 10.1111/j.1365-2672.2004.02285.x.
To investigate the influence of the source of tryptone in the growth medium on the resistance of Escherichia coli to various types of oxidative stress.
Cultures of Escherichia coli MG1655 were grown in Luria-Bertani (LB) medium at 37 degrees C to stationary phase, harvested, and subsequently subjected to various types of oxidative stress. A marked difference in oxidative stress sensitivity was observed depending on the origin of the tryptone in the LB medium used to grow the cultures. Cells harvested from LB containing tryptone from source x (LBx) were more sensitive to inactivation by the superoxide generating compound plumbagin and by t-butyl peroxide, and to growth inhibition by the lactoperoxidase enzyme system, than cells harvested from LB containing tryptone from source y (LBy). By monitoring expression of a panel of stress gene promotors linked to the gfp (green fluorescent protein) gene, and using Delta2-22 alkaline phosphatase as a probe for disulphide bridge formation from protein sulphydryl groups, it was demonstrated that a greater cytoplasmic oxidative stress existed in cells during growth in LBy than in LBx.
Depending on the source of tryptone, bacteria may experience different levels of oxidative stress in tryptone-containing nonselective growth media. Although these levels of oxidative stress are subinhibitory, they may trigger a stress response that makes the bacteria more resistant to a subsequent exposure to a lethal or inhibitory level of oxidative stress.
This work highlights the importance of controlling very subtle differences in composition of nonselective growth media in studies on bacterial physiology.
研究生长培养基中胰蛋白胨的来源对大肠杆菌抵抗各种氧化应激的影响。
将大肠杆菌MG1655培养物在37℃的Luria-Bertani(LB)培养基中培养至稳定期,收获后,随后使其遭受各种氧化应激。根据用于培养细菌的LB培养基中胰蛋白胨的来源,观察到氧化应激敏感性存在显著差异。与从含有来源y的胰蛋白胨的LB(LBy)中收获的细胞相比,从含有来源x的胰蛋白胨的LB(LBx)中收获的细胞对超氧化物生成化合物白花丹素和叔丁基过氧化物的失活以及对乳过氧化物酶系统的生长抑制更敏感。通过监测一组与gfp(绿色荧光蛋白)基因相连的应激基因启动子的表达,并使用Delta2-22碱性磷酸酶作为蛋白质巯基形成二硫键的探针,结果表明,在LBy中生长期间细胞内的细胞质氧化应激比在LBx中更大。
根据胰蛋白胨的来源,细菌在含胰蛋白胨的非选择性生长培养基中可能经历不同程度的氧化应激。尽管这些氧化应激水平是亚抑制性的,但它们可能引发应激反应,使细菌对随后暴露于致死或抑制水平的氧化应激更具抗性。
这项工作突出了在细菌生理学研究中控制非选择性生长培养基组成中非常细微差异的重要性。