1887

Abstract

Summary: A thiosulphate-oxidizing system of grown on thiosulphate was studied using intact cells, crude cell-free extracts and isolated membrane vesicles. All active preparations oxidized thiosulphate to sulphate, without intermediary accumulation of polythionates, consuming two moles of oxygen for every mole of thiosulphate oxidized. Less active cell-free preparations required reduced glutathione, NADH or sulphite for thiosulphate oxidation. An active membrane-associated thiosulphate-oxidizing system from crude cell-free extracts was isolated by either Sepharose 4B column chromatography or differential centrifugation. The isolated multi-enzyme complex system exhibited high specificity for thiosulphate with an apparent value of 0·12 m. Enzyme activity was optimum at pH 7·5 and 25 °C. The system was sensitive to oxygen, storage at certain temperatures and freezing. Inhibition studies indicated that active sulphydryl groups and an electron transport chain were involved during thiosulphate oxidation. Electron micrographs of active crude extracts and the isolated membrane complex after negative staining showed the presence of spherical structures with a diameter of about 0·1 to 0·4 m. An ultra-thin section of the membrane complex revealed that the large spherical particles observed in the negatively stained preparations were aggregated structures consisting of smaller vesicles 0·02 to 0·05 m in diameter.

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/content/journal/micro/10.1099/00221287-99-2-397
1977-04-01
2024-12-06
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References

  1. Bodo C., Lundgren D.G. 1974; Iron oxidation by cell envelopes of Thiobacillus ferrooxidans. Canadian Journal of Microbiology 20:1647–1652
    [Google Scholar]
  2. Bray G.A. 1960; A simple efficient liquid scintillator for counting aqueous solutions in a liquid scintillation counter. Analytical Biochemistry 1:279–285
    [Google Scholar]
  3. Charles A.M., Suzuki I. 1966; Mechanism of thiosulphate oxidation by Thiobacillus novellus. Biochimica et biophysica acta 128:510–521
    [Google Scholar]
  4. Dular U. 1975 Biochemical and structural studies of the cell-free ammonia-oxidizing system of Nitrosomonas europaea Ph.D. thesis University of Manitoba, Canada.:
    [Google Scholar]
  5. Kelly D.P. 1970; Thin-layer chromatography of inorganic sulphur compounds. Journal of Chromatography 51:343–345
    [Google Scholar]
  6. London J., Rittenberg S.G. 1964; Path of S in sulphide and thiosulphate oxidation by thiobacilli. Proceedings of the national academy of sciences of the united states of america 52:1183–1190
    [Google Scholar]
  7. Lowry O.H., Rosebrough N.J., Farr A.L., Randall R.J. 1951; Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193:265–275
    [Google Scholar]
  8. Lyric M.R., Suzuki I. 1970; Enzymes involved in the metabolism of thiosulphate by Thiobacillus thioparus. 1. Survey of enzymes and properties of sulphite:cytochrome c oxidoreductase. Canadian Journal of Biochemistry 48:334–343
    [Google Scholar]
  9. Massey V., Edmondson D. 1970; On the mechanism of inactivation of xanthine oxidase by cyanide. Journal of Biological Chemistry 245:6595–6598
    [Google Scholar]
  10. Oh J.K., Suzuki I. 1977; Resolution of a membrane-associated thiosulphate-oxidizing complex of Thiobacillus noveltus. Journal of General Microbiology 99:1053–1057
    [Google Scholar]
  11. Peck H.D.Jr 1960; Adenosine 5′-phosphosulphate as an intermediate in the oxidation of thiosulphate by Thiobacillus thioparus. Proceedings of the National Academy of Sciences of the United States of America 461053–1057
    [Google Scholar]
  12. Peck H.D.Jr 1968; Energy metabolism in chemolithotrophic bacteria. Annual Review of Microbiology 22:489–518
    [Google Scholar]
  13. Peck H.D.Jr Fisher E.Jr 1962; The oxidation of thiosulphate and phosphorylation on extracts of Thiobacillus thioparus. Journal of Biological Chemistry 237:190–197
    [Google Scholar]
  14. Roy A.B., Trudinger P.A. 1970 The Biochemistry of Inorganic Compounds of Sulphur London: Cambridge University Press.;
    [Google Scholar]
  15. Sörbo B. 1957; A colorimetric method for the determination of thiosulphate. Biochimica et biophysica acta 23:412–416
    [Google Scholar]
  16. Starkey R.L. 1934; Cultivation of organisms concerned in the oxidation of thiosulphate. Journal of Bacteriology 28:365–386
    [Google Scholar]
  17. Suzuki I. 1974; Mechanisms of inorganic oxidation and energy coupling. Annual Review of Microbiology 28:85–101
    [Google Scholar]
  18. Suzuki I., Kwok S.C. 1969; Oxidation of ammonia by spheroplasts of Nitrosomonas europaea. Journal of Bacteriology 99:897–898
    [Google Scholar]
  19. Suzuki I., Kwok S.C. 1970; Cell-free ammonia oxidation by Nitrosomonas europaea extracts: effects of polyamines, Mg2+, and albumin. Biochemical and Biophysical Research Communications 39:950–955
    [Google Scholar]
  20. Suzuki I., Silver M. 1966; The initial product and properties of the sulphur-oxidizing enzyme of thiobacilli. Biochimica et biophysica acta 122:22–33
    [Google Scholar]
  21. Trudinger P.A. 1964a; Evidence for a four-sulphur intermediate in thiosulphate oxidation by Thiobacillus X. Australian Journal of Biological Science 17:577–579
    [Google Scholar]
  22. Trudinger P.A. 1964b; Oxidation of thiosulphate by intact cells of Thiobacillus X. Effects of some experimental conditions. Australian Journal of Biological Science 17:738–751
    [Google Scholar]
  23. Trudinger P.A. 1964c; The effects of thiosulphate and oxygen concentration on tetrathionate oxidation by Thiobacillus X and T. thioparus. Biochemical Journal 90:640–646
    [Google Scholar]
  24. Vishniac W., Trudinger P.A. 1962; Carbon dioxide fixation and substrate oxidation in the chemosynthetic sulphur and hydrogen bacteria. Bacteriological Reviews 26:168–175
    [Google Scholar]
  25. Westley J. 1973; Rhodanese. Advances in Enzymology 39:327–368
    [Google Scholar]
  26. Wharton D.C., Tzagoloff A. 1967; Cytochrome oxidase from beef heart mitochondria. Methods in Enzymology 10:245–246
    [Google Scholar]
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