1887

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Volume 143, Issue 5

A 16 kDa protein family overexpressed by PB18 in acid environments Free

Abstract

The one- and two-dimensional protein patterns of PB18 in the exponential and stationary phases of growth were analysed. Onedimensional SDS-PAGE showed that a 16 kDa protein was overexpressed in stationary phase as well as 2 h after an acid shock, and that it was not expressed when the bacteria reached the stationary phase in medium with limiting lactose concentrations (5 or 10 g l), in which the pH (5∙5) was not as acid as in control cultures (pH 4∙7, lactose 20 g l). The results support the idea that this protein is expressed in response to the acidic environment and not in response to the growth phase. Two-dimensional PAGE showed that nine proteins were expressed only during the exponential phase and ten others only during the stationary phase. The 16 kDa band seen in one-dimensional SDS-PAGE corresponded to a 16 kDa protein family observed on two-dimensional SDS-PAGE/IEF gels, whose expression was increased 8∙5-fold when the extracellular pH reached a critical value below 5∙0. The N-terminal sequences of proteins from two spots on the two-dimensional gels (members of the 16 kDa family) were determined and found to be identical. The physiological role of this protein family has not yet been elucidated.

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Keyword(s): 2D electrophoresis , acid stress , intracellular pH and Streptococcus thermophilus
© Society Society for General Microbiology 1997
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1997-05-01
2024-04-19
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References

  1. Altschul S. F., Gish W., Miller W., Meyers E. Z., Lipman D. J. 1990; Basic local alignment search tool. J Mol Biol 215:403–410
     [Google Scholar]
  2. Auffray Y., Lecesne E., Hartke A., Boutibonnes P. 1995; Basic features of the Streptococcus thermophilus heat shock response. Curr Microbiol 30:87–91
     [Google Scholar]
  3. Baumberg S., Klingel U. 1993; Biosynthesis of arginine, proline, and related compounds. In Bacillus subtilis and Other Gram-positive Bacteria, Edited by A. L. Sonenshein, R. Losick & J. A. Hoch. Washington, DC:. American Society for Microbiology.299–306
     [Google Scholar]
  4. Booth I. R. 1985; Regulation of cytoplasmic pH in bacteria. Microbiol Rev 49:359–378
     [Google Scholar]
  5. Bracquart P. 1981; An agar medium for the differential enumeration of Streptococcus thermophilus and Lactobacillus bulgaricus in yoghurt. J Appl Bacteriol 51:303–305
     [Google Scholar]
  6. Bradford M. M. 1976; A rapid and sensitive method for the quantification of microgram quantities of protein utilising the principle of protein dye binding. Anal Biochem 72:248–254
     [Google Scholar]
  7. Foster J. W., Hall H. K. 1991; Inducible pH homeostasis and the acid tolerance response of Salmonella typhimurium. J Bacteriol 173:5129–5135
     [Google Scholar]
  8. Groat R. G., Hall H. K., Schultz J. E., Zychlinsky E., Bockman A., Matin A. 1986; Starvation proteins in Escherichia coli: kinetics of synthesis and role in starvation. J Bacteriol 168:486–493
     [Google Scholar]
  9. Guimont C., Clary D., Bracquart P. 1994; Analysis of wholecell proteins of Streptococcus thermophilus by two electrophoretic methods. Lait 74:13–21
     [Google Scholar]
  10. Guzzo J., Cavin J. F., Divies C. 1994; Induction of stress proteins in Leuconostoc oenos to permit direct inoculation of wine. Biotechnol Lett 16:1189–1194
     [Google Scholar]
  11. Hamilton I. R., Buckley N. D. 1991; Adaptation by Streptococcus mutans to acid tolerance. Oral Microbiol Zmmunot 6:65–71.
     [Google Scholar]
  12. Hashiba H., Takiguchi R., Joho K., Aoyama K., Hirota T. 1993; Identification of the replication region of Streptococcus thermophilus No. 29 plasmid pST1. Biosci Biotechnol Biochem 57:1646–1649
     [Google Scholar]
  13. Hengge-Aronis R. 1993; Survival of hunger and stress: the role of rpoS in early stationary phase gene regulation in E . coli. Cell 72:165–168
     [Google Scholar]
  14. Hickey E. W., Hirshfield I. N. 1990; Low-pH-induced effects on patterns of protein synthesis and on internal pH in Escherichia coli and Salmonella typhimurium. Appl Environ Microbiol 56:1038–1045
     [Google Scholar]
  15. Hutkins R. W., Nannen N. L. 1993; pH homeostasis in lactic acid bacteria. J Dairy Sci 76:2354–2365.
     [Google Scholar]
  16. Kashket E. R. 1985; The proton motive force in bacteria: a critical assessment of methods. Annu Rev Microbiol 39:219–242
     [Google Scholar]
  17. Kashket E. R. 1987; Bioenergetics of lactic acid bacteria: cytoplasmic pH and osmotolerance. FEMS Microbiol Rev 46:233–244
     [Google Scholar]
  18. Kolter R., Siegele D. A., Tormo A. 1993; The stationary phase of the bacterial life cycle. Annu Rev Microbiol 47:855–874
     [Google Scholar]
  19. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
     [Google Scholar]
  20. Lee I. S., Lin J., Hall H. K., Bearson B., Foste J. W. 1995; The stationary-phase sigma factor σS (RpoS) is required for a sustained acid tolerance response in virulent Salmonella typhimurium. Mol Microbiol 17:155–167
     [Google Scholar]
  21. Nannen N. L., Hutkins R. W. 1991; Intracellular pH effects in lactic acid bacteria. J Dair Sci 74:741–746
     [Google Scholar]
  22. O’Farrell P. H. 1975; High-resolution two-dimensional electrophoresis of proteins. J Biol Chem 250:4007–4021
     [Google Scholar]
  23. Patton Z. F., Pluskal M. G., Skeq W. M., Buecker J. L., Lopez M. L., Zimmermann R., Belanger L. M., Hatch P. 1990; Development of a dedicated two-dimensional gel electrophoresis system that provides optimal pattern reproducibility and polypeptide resolution. Biotechniques 8:518–527
     [Google Scholar]
  24. Piard J. C, Desmazeaud M. 1991; Inhibiting factors produced by lactic acid bacteria. 1. Oxygen metabolites and catabolism end-products. Lait 71:525–541
     [Google Scholar]
  25. Rallu F., Gruss A., Maguin E. 1996; Lactococcus lactis and stress. Antonie Leeuwenhoek 70:243–251
     [Google Scholar]
  26. Sauer U., Durre P. 1993; Sequence and molecular characterization of a DNA region encoding a small heat shock protein of Clostridium acetobutylicum. J Bacteriol 175:3394–3400
     [Google Scholar]
  27. Servant P., Mazodier P. 1995; Characterization of Streptomyces albus 18-kilodalton heat shock-responsive protein. J Bacteriol 177:2998–3003
     [Google Scholar]
  28. Sonenshein A. L. 1989; Metabolic regulation of sporulation and other stationary phase phenomena. In Regulation of Procaryotic Development, Edited by I. Smith, R. A. Slepecky & P. Setlow. Washington, DC:. American Society for Microbiology.109–130
     [Google Scholar]
  29. Terzaghi B. E., Sandine W. 1975; Improved medium for lactic streptococci and their bacteriophages. Appl Microbiol 29:807–813
     [Google Scholar]
  30. Towbin H., Staehelin T., Gordon J. 1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets : procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354
     [Google Scholar]
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A 16 kDa protein family overexpressed by Streptococcus thermophilus PB18 in acid environments
Microbiology 143, 1587 (1997); https://doi.org/10.1099/00221287-143-5-1587
/content/journal/micro/10.1099/00221287-143-5-1587
/content/journal/micro/10.1099/00221287-143-5-1587
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