1887

Abstract

Summary: A method was established to measure the cytoplasmic pH of the facultative alkaliphilic strain, . The bacterium was loaded with a pH-sensitive fluorescent probe, 2′,7′-bis-(2-carboxyethyl)-5 (and -6)-carboxyfluorescein (BCECF), and cytoplasmic pH was determined from the intensity of fluorescence of the intracellular BCECF. The activity of the organism to maintain neutral cytoplasmic pH was assessed by measuring the cytoplasmic pH of the cells exposed to various pH conditions. The cytoplasmic pH maintenance activity of C-125 increased with increasing culture pH, indicating that the activity was regulated in response to the culture pH.

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/content/journal/micro/10.1099/00221287-143-8-2531
1997-08-01
2021-05-09
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References

  1. Aono R. 1985; Isolation and partial characterization of structural components of the walls of alkalophilic Bacillus strain C-125. J Gen Microbiol 131:105–111
    [Google Scholar]
  2. Aono R. 1995; Assignment of facultatively alkaliphilic Bacillus sp. C-125 to Bacillus lentus group 3. Int J Syst Bacteriol 45:582–585
    [Google Scholar]
  3. Aono R., Horikoshi K. 1983; Chemical composition of cell walls of alkalophilic strains of Bacillus. . J Gen Microbiol 129:1083–1087
    [Google Scholar]
  4. Aono R., Sanada T. 1994; Hyper-autolysis of the facultative alkaliphile Bacillus sp. C-125 cells grown at neutral pH: culture-pH-dependent cross-linking of the peptide moieties of the peptidoglycan. Biosci Biotechnol Biochem 58:2015–2019
    [Google Scholar]
  5. Aono R., Ito M., Joblin K. N., Horikoshi K. 1995; A high cell wall negative charge is necessary for the growth of the alkaliphile Bacillus lentus C-125 at elevated pH. Microbiology 141:2955–2964
    [Google Scholar]
  6. Aono R., Kaneko H., Horikoshi K. 1996; Alkaline growth pH-dependent increase of respiratory and NADH oxidation activities of facultatively alkaliphilic strain Bacillus lentus C-125. Biosci Biotechnol Biochem 60:1243–1248
    [Google Scholar]
  7. Dencher N. A., Burghaus P. A., Grzesiek S. 1986; Determination of the net proton-hydroxide ion permeability across vesicular lipid bilayers and membranes proteins by optical probes. Methods Enzymol 127:746–760
    [Google Scholar]
  8. Futsaether C. M., Kjeldstad B., Johnsson A. 1993; Measurement of the intracellular pH of Propionibacterium acnes: comparison between the fluorescent probe BCECF and 31P-NMR spectroscopy. Can J Microbiol 39:180–186
    [Google Scholar]
  9. Hashimoto M., Hamamoto T., Kitada M., Hino M., Kudo T., Horikoshi K. 1994; Characteristics of alkali-sensitive mutants of alkaliphilic Bacillus sp. strain C-125 that show cellular morphological abnormalities. Biosci Biotechnol Biochem 58:2090–2092
    [Google Scholar]
  10. Kitada M., Onda K., Horikoshi K. 1989; The sodium/proton antiport system in a newly isolated alkalophilic Bacillus sp. J Bacteriol 171:1879–1884
    [Google Scholar]
  11. Kitada M., Hashimoto M., Kudo T., Horikoshi K. 1994; Properties of two different Na+/H+ antiport systems in alkaliphilic Bacillus sp. strain C-125. J Bacteriol 176:6464–6469
    [Google Scholar]
  12. Krulwich T. A. 1995; Alkaliphiles: ‘ basic’ molecular problems of pH tolerance and bioenergetics. Mol Microbiol 15:403–410
    [Google Scholar]
  13. Miller J. H. 1972 Experiments in Molecular Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  14. Molenaar D., Abee T., Konings W. N. 1991; Continuous measurement of the cytoplasmic pH in Lactococcus lactis with a fluorescent pH indicator. Biochim Biophys Acta 1115:75–83
    [Google Scholar]
  15. Molenaar D., Bolhuis H., Abee T., Konings W. N. 1992; The efflux of a fluorescent probe is catalyzed by an ATP-driven extrusion system in Lactococcus lactis. . J Bacteriol 174:118–124
    [Google Scholar]
  16. Padan E., Schuldiner S. 1986; Intracellular pH regulation in bacterial cells. Methods Enzymol 125:337–352
    [Google Scholar]
  17. Sturr M. G., Guffanti A. A., Krulwich T. A. 1994; Growth and bioenergitics of alkaliphilic Bacillus firmus OF4 in continuous culture at high pH. J Bacteriol 176:3111–3116
    [Google Scholar]
  18. Sugiyama S., Matsukura H., Imae Y. 1985; Relationship between Na+-dependent cytoplasmic pH homeostasis and Na+- dependent flagella rotation and amino acid transport in alkalophilic Bacillus. . FEBS Lett 182:265–268
    [Google Scholar]
  19. Tsuchiya T., Takeda K. 1979; Calcium/proton and sodium/proton antiport systems in Escherichia coli. . J Biochem 85:943–951
    [Google Scholar]
  20. Tsujimoto K., Semadeni M., Huflejt M., Packer L. 1988; Intracellular pH of halobacteria can be determined by the fluorescent dye 2ʹ,7ʹ-bis-(2-carboxyethyl)-5(6)-carboxy-fluorescein. Biochem Biophys Res Commun 155:123–129
    [Google Scholar]
  21. Zilverstein D., Agmon V., Schuldiner S., Padan E. 1982; The sodium/proton antiporter is part of the pH homeostasis mechanism in Escherichia coli. . J Biol Chem 257:3687–3691
    [Google Scholar]
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