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

Abstract

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 

Abbreviations

BVH

reduced benzyl viologen

CCCP

carbonyl cyanide m-chlorophenyl hydrazone

DIECA

N, N-diethyl-dithiocarbamate

HOQNO

2-n-heptyl-4-hydroxyquinoline-N-oxide

NEM

N-ethylmaleimide

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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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