Archives of Microbiology

, Volume 112, Issue 2, pp 163–168 | Cite as

Acidostability of spheroplasts prepared from Thiobacillus thiooxidans

  • Akinorl Noguchi
  • Michihiro Takama
  • Takeshi Sekiguchi
  • Noriyuki Koyama
  • Yoshiaki Nosoh


Thiobacillus thiooxidans was acidostable even in the absence of its respiratory substrate, elementary sulfur. This suggests that the acidostability of the bacterium was energy-independent. The organism was subjected to osmotic shock with 0.75 M sucrose at 0°C and then treated with snail intestinal juice in the presence of 0.3 M sucrose. The decrease in the optical density of the sample thus prepared on dilution with deionized water and electron microscopic observation of the sample showed that spheroplasts were formed from the bacterium by this procedure. Spheroplasts were able to respire sulfur and their respiratory activity was acidostable. Spheroplasts, when treated with Nagase, proteolytic enzyme, lost their acidostability, and some protein components disappeared from the membrane fraction. This suggests that the acidostability of the bacterium may be related to protein components of the membrane.

Key words

Thiobacillus thiooxidans Sulfur oxidizing bacterium Loss of acidostability Spheroplast formation Snail intestinal juice Osmotic shock Proteinase treatment Membrane proteins 


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  1. Anderson, F. B., Millbank, J. W.: Protoplast formation and yeast cell-wall structure. The action of the enzyme of the snail, Helix pomatia. Biochem. J. 99, 682–687 (1966)Google Scholar
  2. Barridge, J. K., Shively, J. M.: Phospholipids of the Thiobacilli. J. Bact. 95, 2182–2185 (1968)Google Scholar
  3. Birdsell, D. C., Cota-Robles, E. H.: Production and ultrastructure of lysozyme and ethylenediaminetetraacetate-lysozyme spheroplasts of Escherichia coli. J. Bact. 93, 427–437 (1967)Google Scholar
  4. Brock, T. D.: Microbial growth under extreme conditions. In: Microbial growth (P. M. Meadow, S. U. Pirst, eds.), pp. 15–41. Cambridge: Cambridge University Press 1969Google Scholar
  5. Brock, T. D., Brock, K. M., Belly, R. T., Weiss, R. Z.: Sulfolobus: A new genus of sulfur-oxidizing bacteria at low pH and high temperature. Arch. Mikrobiol. 84, 54–68 (1972)Google Scholar
  6. Crum, E. H., Siehr, D. J.: Thiobacillus thiooxidans cell wall amino acids and monosaccharides. J. Bact. 94, 2069–2070 (1967)Google Scholar
  7. Darland, G., Brock, T. D.: Bacillus acidocalarius sp. nov., an acidophilic thermophilic sporeforming bacterium. J. gen. Microbiol. 67, 9–15 (1971)Google Scholar
  8. Eddy, A. A., Williamson, D. H.: A method of isolating protoplasts from yeast. Nature (Lond.) 179, 1252–1253 (1957)Google Scholar
  9. Fairbanks, G., Steck, T. L., Wallach, D. F. H.: Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry 10, 2606–2616 (1971)Google Scholar
  10. Folch, J., Lees, M., Sloane-Stanley, G. H.: A simple method for the isolation and purification of total lipids from animal tissues. J. biol. Chem. 226, 497–509 (1957)Google Scholar
  11. Haest, C. W. M., Degier, J., Op den Kamp, J. A. F., Bartels, P., Van Deenen, L. L. M.: Changes in permeability of Staphylococcus aureus and derived liposome with varying lipid composition. Biochim. biophys. Acta (Amst.) 255, 720–733 (1972)Google Scholar
  12. Iwatsuka, H., Kano, M., Maruyama, M.: Metabolism of sulfuroxidizing bacteria. II. System of CO2-fixation in Thiobacillus thiooxidans. Plant Cell Physiol. 3, 157–166 (1962)Google Scholar
  13. Kodama, A.: Studies on the metabolism of a sulfur-oxidizing bacterium. VI. Fractionation and reconstitution of the elementary sulfur-oxidizing system of Thiobacillus thiooxidans. Plant Cell Physiol. 10, 645–655 (1969)Google Scholar
  14. Koyama, N., Nosoh, Y.: Effect of the pH of culture medium on the alkalophilicity of a species of Bacillus. Arch. Microbiol. 109, 105–108 (1976)Google Scholar
  15. Levin, R. A.: Fatty acids of Thiobacillus thiooxidans. J. Bact. 108, 992–995 (1971)Google Scholar
  16. Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J.: Protein measurement with the Folin phenol reagent. J. biol. Chem. 193, 265–275 (1951)Google Scholar
  17. Luft, J. H.: Improvements in epoxy resin embedding methods. J. biophys. biochem. Cytol. 9, 409–414 (1961)Google Scholar
  18. Mahoney, R. P., Edwards, M. R.: Fine structure of Thiobacillus thiooxidans. J. Bact. 92, 487–495 (1966)Google Scholar
  19. Marunouchi, T., Mori, T.: The nature of the cell surface in Thiobacillus thiooxidans, a solid sulfur oxidizing bacterium. Bot. Mag. Tokyo 81, 179–189 (1968)Google Scholar
  20. Miura, T., Mizushima, S.: Separation and properties of outer and cytoplasmic membranes in Escherichia coli. Biochim. biophys. Acta (Amst.) 193, 268–276 (1969)Google Scholar
  21. Ohta, K., Kiyomiya, A., Koyama, N., Nosoh, Y.: The basis of the alkalophilic property of a species of Bacillus. J. gen. Microbiol. 86, 259–266 (1975)Google Scholar
  22. Osborn, M. J., Gander, J. E., Parisi, E., Carson, J.: Mechanism of assembly of the outer membrane of Salmonella typhimurium. Isolation and characterization of cytoplasmic and outer membrane. J. biol. Chem. 247, 3962–3972 (1972)Google Scholar
  23. Oshima, M., Ariga, T.: ω-Cyclohexyl fatty acids in acidophilic thermophilic bacteria. Studies on their presence, structure and biosynthesis using precursors labelled with stable isotopes and radioisotopes. J. biol. Chem. 250, 6963–6968 (1975)Google Scholar
  24. Phuff, H. J.: The yeasts. New York-London: Academic Press 1971Google Scholar
  25. Ryter, A., Kellenberger, E.: Etude au microscope électronique de plasma contenant de l'acide deoxyribonucléique. Z. Naturforsch. 13b, 597–605 (1958)Google Scholar
  26. Sabatini, D. D., Bensch, K., Barnett, R. J.: Cytochemistry and electron microscopy: The preservation of cellular ultrastructure and enzymic activity by aldehyde fixation. J. Cell Biol. 17, 19–58 (1963)Google Scholar
  27. Suzuki, I.: Oxidation of elementary sulfur by an enzyme system of Thiobacillus thiooxidans. Biochim. biophys. Acta (Amst.) 104, 359–371 (1965)Google Scholar
  28. Uchino, F., Doi, S.: Acido-thermophilic bacteria from thermal waters. Agr. biol. Chem. 31, 817–822 (1967)Google Scholar
  29. Venable, J. H., Coggeshall, R.: A simplified lead citrate stain for use in electron microscopy. J. Cell Biol. 25, 407–408 (1965)Google Scholar
  30. Vishniac, W., Santer, M.: The thiobacilli. Bact. Rev. 21, 195–213 (1957)Google Scholar
  31. Yamazaki, Y., Koyama, N., Nosoh, Y.: On the acidostability of an acidophilic thermophilic bacterium. Biochim. biophys. Acta (Amst.) 314, 257–260 (1973)Google Scholar

Copyright information

© Springer-Verlag 1977

Authors and Affiliations

  • Akinorl Noguchi
    • 1
  • Michihiro Takama
    • 1
  • Takeshi Sekiguchi
    • 1
  • Noriyuki Koyama
    • 1
  • Yoshiaki Nosoh
    • 1
  1. 1.Laboratory of Chemistry of Natural ProductsTokyo Institute of TechnologyTokyoJapan

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