1887

Abstract

The lipophilic cations butyltriphenylphosphonium (BTPP), tetraphenylphosphonium (TPP) and triphenylmethylphosphonium (TPMP) were taken up into cells of H18 under anaerobic conditions. Uptake was dependent on the presence of maltose together with both HCO and Na; it was at a maximum at concentrations of ≥ 20 m-HCO and ≥2 m-Na. The addition of 2-(n-heptyl)-hydroxyquinoline--oxide (HpHOQnO) or of an uncoupler of oxidative phosphorylation, or air, resulted in efflux of the lipophilic cations. From the binding behaviour and the physiological effects of the lipophilic cations, a membrane potential (Δψ) of 140 mV was estimated. There was no detectable ΔpH at an external pH of 6·7. The cytoplasmic Naconcentration was estimated to be 0·2 m, indicating that can maintain a Naconcentration gradient equivalent to at least 150 mV. Variation in the external Naconcentration (2 to 180 m) had little influence on Δψ. High external concentrations of the fermentation products acetate, formate and succinate had little effect on the growth rate and the Δψ. The cytoplasmic ATP concentration decreased rapidly on addition of HpHOQnO oxide or of an uncoupler. The maximum internal ATP concentration was only maintained at an external concentration≥2m-Na. The NADH: fumarate reductase activity of vesicles of was associated with alkalization of the suspension medium. The amount of Htaken up was in excess of the expected amount of scalar Hand was partially sensitive to uncoupler. It is concluded that the Δψ was generated via Htranslocation driven primarily by the cytochrome-deficient NADH: fumarate reductase system. The transmembrane Nagradient could be supported via the action of a Na/2 Hantiporter.

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1987-01-01
2024-04-18
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References

  1. Alefounder P. R., Mccarthy J. E. G., Ferguson S. J. 1981; The basis of the control of nitrate reduction by oxygen in Paracoccus denitrificans. FEMS Microbiology Letters 12:321–326
    [Google Scholar]
  2. Blaut M., Müller V., Fiebig K., Gottschalk G. 1985; Sodium ions and an energized membrane are required by Methanosarcina barkeri for the oxidation of methanol to the level of formaldehyde. Journal of Bacteriology 164:95–101
    [Google Scholar]
  3. Caldwell D. R., Kenney M., Barton J. S., Kelley J. F. 1973; Sodium and other inorganic growth requirements of Bacteroides amylophilus. Journal of Bacteriology 114:782–789
    [Google Scholar]
  4. Faust P. J., Vandemark P. J. 1970; Phosphorylation coupled to NADH oxidation with fumarate in Streptococcus faecalis 10 Cl. Archives of Biochemistry and Biophysics 137:392–398
    [Google Scholar]
  5. Gest H. 1980; The evolution of biological energy- transducing systems. FEMS Microbiology Letters 7:73–77
    [Google Scholar]
  6. Harold F. M., Heefner D. L. 1981; ATP and proton-motive force in bacterial ion transport: variations on a theme by Mitchell. In Chemiosmotic Proton Circuits in Biological Membranes pp. 537–550 Skulachev V. P., Hinkle P. C. Edited by Reading, Mass., USA: Addison-Wesley;
    [Google Scholar]
  7. Hellingwerf K. J., Bolscher J. G. M., Konings W. N. 1981; The electrochemical proton gradient generated by the fumarate-reductase system in Escherichia coli and its bioenergetic implications. European Journal of Biochemistry 113:369–374
    [Google Scholar]
  8. Hilpert W., Schink B., Dimroth P. 1984; Life by a new decarboxylation-dependent energy conservation mechanism with Na+ as coupling ion. EMBO Journal 3:1665–1670
    [Google Scholar]
  9. Hobson P. N., Summers R. 1967; The continuous culture of anaerobic bacteria. Journal of General Microbiology 47:53–65
    [Google Scholar]
  10. Hobson P. N., Wallace R. J. 1982; Microbial ecology and activities in the rumen. Part II. Critical Reviews in Microbiology 9:253–320
    [Google Scholar]
  11. Jenkinson H. F., Woodbine M. 1979; Growth and energy production in Bacteroides amylophilus. Archives of Microbiology 120:275–281
    [Google Scholar]
  12. Klein W. L., Boyer P. D. 1972; Energization of active transport by Escherichia coli. Journal of Biological Chemistry 247:7257–7265
    [Google Scholar]
  13. Konings W. N., Michels P. A. M. 1980; Electron transfer-driven solute translocation across bacterial membranes. In Diversity of Bacterial Respiratory Systems 1 pp. 33–86 Knowles C. J. Edited by Boca Raton, Fla., USA: CRC Press;
    [Google Scholar]
  14. Kröger A. 1978; Fumarate as terminal acceptor of phosphorylative electron transport. Biochimica et biophysica acta 505:129–145
    [Google Scholar]
  15. Lesk E. M., Blackburn T. H. 1971; Purification of Bacteroides amylophilus protease. Journal of Bacteriology 106:394–402
    [Google Scholar]
  16. Lolkema J. S., Hellingwerf K. J., Konings W. N. 1982; The effect of “probe binding” on the quantitative determination of the proton motive force in bacteria. Biochimica et biophysica acta 681:85–94
    [Google Scholar]
  17. Macy J., Probst I. A., Gottschalk G. 1975; Evidence for cytochrome involvement in fumarate reduction and adenosine-5′-triphosphate synthesis by Bacteroides fragilis grown in the presence of hemin. Journal of Bacteriology 123:436–442
    [Google Scholar]
  18. Maloney P. C. 1983; Relationship between phosphorylation potential and electrochemical H+ gradient during glycolysis in Streptococcus lactis. Journal of Bacteriology 153:1461–1470
    [Google Scholar]
  19. Muratsugu M., Kamo N., Kobatake Y., Kimura K. 1979; Determination of membrane potential for Escherichia coli with use of an electrode sensitive to tetraphenyl phosphonium. Journal of Electroanalytical Chemistry 104:477–491
    [Google Scholar]
  20. Mccarthy J. E. G., Ferguson S. J. 1983; Characterisation of membrane vesicles from Paracoccus denitrificans and measurement of the effect of partial uncoupling on their thermodynamics of oxidative phosphorylation. European Journal of Biochemistry 132:417–424
    [Google Scholar]
  21. Mitchell P. 1966; Chemiosmotic coupling in oxidative and photosynthetic phosphorylation. Biological Reviews 41:445–502
    [Google Scholar]
  22. Padan E., Zllberstein D., Schuldiner S. 1981; pH homeostasis in bacteria. Biochimica et biophysica acta 650:151–166
    [Google Scholar]
  23. Perski H. J., Moll J., Thauer R. K. 1981; Sodium dependence of growth and methane formation in Methanobacterium thermoautotrophicum. Archives of Microbiology 130:319–321
    [Google Scholar]
  24. Ramos S., Schuldiner S., Kaback H. R. 1979; The use of flow dialysis for determination of ApH and active transport. Methods in Enzymology 55:680–688
    [Google Scholar]
  25. Reddy C. A., Bryant M. P. 1977; Deoxyribonucleic acid base composition of certain species of the genus Bacteroides. Canadian Journal of Microbiology 23:1252–1256
    [Google Scholar]
  26. Schienz K. L., Rütten B., Tretter M. 1981; Determination of adenosine nucleotides with luciferin/luciferase from crude firefly lantern extract on a bioluminescence analyzer. Advances in Biotechnology 1:457–462
    [Google Scholar]
  27. Scholes P., Mitchell P. 1970; Respiration-driven proton translocation in Micrococcus denitrificans. Bioenergetics 1:309–323
    [Google Scholar]
  28. Sorgato M. C., Ferguson S. J., Kell D. B., John P. 1978; The proton-motive force in bovine heart submitochondrial particles. Magnitude, sites of generation and comparison with the phosphorylation potential. Biochemical Journal 174:237–256
    [Google Scholar]
  29. Stouthamer A. H. 1980; Electron linked phosphorylation in anaerobes. In Anaerobes and Anaerobic Infections pp. 17–29 Gottschalk G., Pfennig N., Werner H. Edited by Stuttgart: Fischer;
    [Google Scholar]
  30. Taussky H. H., Shorr E. 1953; A microcolorimetric method for the determination of inorganic phosphorus. Journal of Biological Chemistry 202:675–685
    [Google Scholar]
  31. Ten Brink B., Konings W. N. 1982; Electrochemical proton gradient and lactate concentration gradient in Streptococcus cremoris cells grown in batch culture. Journal of Bacteriology 152:682–686
    [Google Scholar]
  32. Thauer R. K., Jungermann K., Decker K. 1977; Energy conservation in chemotrophic anaerobic bacteria. Bacteriological Reviews 41:100–180
    [Google Scholar]
  33. Tokuda H., Unemoto T. 1982; Characterization of the respiration-dependent Na+ pump in the marine bacterium Vibrio alginolyticus. Journal of Biological Chemistry 257:10007–10014
    [Google Scholar]
  34. Vogel G., Steinhart R. 1976; ATPase of Escherichia coli: purification, dissociation and reconstitution of the active complex from the isolated subunits. Biochemistry 15:208–216
    [Google Scholar]
  35. Wetzstein H. G., Gottschalk G. 1985; A sodium-stimulated membrane-bound fumarate reductase system in Bacteroides amylophilus. Archives of Microbiology 143:157–162
    [Google Scholar]
  36. Wilson T. H., Lin E. C. C. 1980; Evolution of membrane bioenergetics. Journal of Supramolecular Structure 13:421–446
    [Google Scholar]
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