Hickey E W, Hirshfield I N
Department of Biological Sciences, St. John's University, Jamaica, New York 11439.
Appl Environ Microbiol. 1990 Apr;56(4):1038-45. doi: 10.1128/aem.56.4.1038-1045.1990.
Escherichia coli and Salmonella typhimurium were grown in a supplemented minimal medium (SMM) at a pH of 7.0 or 5.0 or were shifted from pH 7.0 to 5.0. Two-dimensional gel electrophoretic analysis of proteins labeled with H2(35)SO4 for 20 min during the shift showed that in E. coli, 13 polypeptides were elevated 1.5- to 4-fold, whereas in S. typhimurium, 19 polypeptides were increased 2- to 14-fold over the pH 7.0 control. Upon long-term growth at pH 5.0, almost double the number of polypeptides were elevated twofold or more in S. typhimurium compared with E. coli. In E. coli, there was no apparent induction of heat shock proteins upon growth at pH 5.0 in SMM. However, growth of E. coli in a complex broth to pH 5.0, or subsequent growth of fresh E. coli cells in the filtrate from this culture, showed that a subset of five polypeptides is uniquely induced by low pH. Two of these polypeptides, D60.5, the inducible lysyl-tRNA synthetase, and C62.5, are known heat shock proteins. Measurements of the internal pH (pHi) and growth rates of both organisms were made during growth in SMM at pH 7.0, pH 5.0, and upon the pH shift. The data show that the pHi of E. coli decreases more severely than that of S. typhimurium at an external pH of 5.0; the growth rate of E. coli is about one-half that of S. typhimurium at this pH, whereas the two organisms have the same growth rate at pH 7.0. The two-dimensional gel, growth, and pHi experiments collectively suggest that, at least in SMM, S. typhimurium is more adaptive to low-pH stress than is E. coli.
大肠杆菌和鼠伤寒沙门氏菌在添加了营养成分的基本培养基(SMM)中,于pH值为7.0或5.0的条件下培养,或者从pH 7.0转变为pH 5.0。在pH值转变过程中,用H2(35)SO4标记蛋白质20分钟后进行二维凝胶电泳分析,结果显示,在大肠杆菌中,有13种多肽的含量升高了1.5至4倍,而在鼠伤寒沙门氏菌中,与pH 7.0的对照相比,有19种多肽的含量增加了2至14倍。在pH 5.0条件下长期生长时,鼠伤寒沙门氏菌中含量升高两倍或更多的多肽数量几乎是大肠杆菌的两倍。在大肠杆菌中,在SMM中于pH 5.0生长时,未明显诱导热休克蛋白。然而,大肠杆菌在复合肉汤中生长至pH 5.0,或者新鲜的大肠杆菌细胞在该培养物的滤液中后续生长,结果表明有一组五种多肽是由低pH值独特诱导产生的。其中两种多肽,即可诱导的赖氨酰 - tRNA合成酶D60.5和C62.5,是已知的热休克蛋白。在SMM中于pH 7.0、pH 5.0以及pH值转变过程中,对这两种微生物的内部pH值(pHi)和生长速率进行了测量。数据表明,在外部pH值为5.0时,大肠杆菌的pHi下降比鼠伤寒沙门氏菌更严重;在此pH值下,大肠杆菌的生长速率约为鼠伤寒沙门氏菌的一半,而在pH 7.0时,这两种微生物的生长速率相同。二维凝胶、生长和pHi实验共同表明,至少在SMM中,鼠伤寒沙门氏菌比大肠杆菌更能适应低pH胁迫。