Archives of Microbiology

, Volume 140, Issue 4, pp 358–364 | Cite as

Proton translocation during denitrification in Rhodopseudomonas sphaeroides f. denitrificans

  • Balaram Kundu
  • D. J. D. Nicholas
Original Papers


Proton translocation during the reduction of NO 3 - , NO 2 - , N2O and O2, with endogenous substrates, in washed cells of Rhodopseudomonas sphaeroides f. denitrificans was investigated by an oxidant pulse method. On adding NO 2 - to washed cells, anaerobically in the dark, an alkalinization occurred in the reaction mixture followed by acidification. When NO 3 - , N2O or O2 was added to cells in the dark or with these compounds and NO 2 - in light an acidification only was observed. Proton translocation was inhibited by carbonyl cyanide-m-chlorophenyl hydrazone.

Valinomycin treated cells produced acid in response to the addition of either NO 3 - , NO 2 - , N2O or O2. The proton extrusion stoichiometry (\(\vec H^ + /2e^ - \) ratios) in illuminated cells were as follows: NO 3 - →0.5N2, 4.82; NO 2 - →0.5N2, 5.43; N2O→N2, 6.20; and O2→H2O, 6.43. In the dark the comparable values were 3.99, 4.10, 4.17 and 3.95. Thus, illuminated cells produced higher \(\vec H^ + /2e^ - \) values than those in the dark, indicating a close link between photosynthesis and denitrification in the generation of proton gradients across the bacterial cell membranes.

When reduced benzyl viologen was the electron donor in the presence of 1 mM N-ethylmaleimide and 0.5 mM 2-n-heptyl-4-hydroxyquinoline-N-oxide in the dark, the addition of either NO 3 - , NO 2 - or N2O to washed cells resulted in a rapid alkalinization of the reaction mixture. The stoichiometries for proton consumption, \(\vec H^ + /2e^ - \) ratios without a permeant ion were NO 3 - →NO 2 - ,-1.95; NO 2 - →0.5 N2O,-3.03 and N2O→N2,-2.02. The data indicate that these reductions occur on the periplasmic side of the cytoplasmic membrane.

Key words

Denitrification Proton translocation Photosynthesis Rhodopseudomonas sphaeroides f. denitrificans 



reduced benzyl viologen


carbonyl cyanide m-chlorophenyl hydrazone


N, N-diethyl-dithiocarbamate






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Copyright information

© Springer-Verlag 1985

Authors and Affiliations

  • Balaram Kundu
    • 1
  • D. J. D. Nicholas
    • 1
  1. 1.Department of Agricultural Biochemistry, Waite Agricultural Research InstituteUniversity of AdelaideGlen OsmondAustralia

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