Proton Translocation Associated with Denitrification in a Photodenitrifier, Rhodopseudomonas sphaeroides forma sp. denitrificans |
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Authors: | Urata Katsuro; Shimada Keizo; Satoh Toshio |
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Institution: | Department of Biology, Tokyo Metropolitan University Fukazawa 2-1-1, Setagaya-ku, Tokyo 158, Japan |
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Abstract: | H+ translocation driven by NO3, NO2 and N2O reductionswith endogenous substrates in cells of Rhodopseudomonas sphaeroidesforma sp. denitrificans was investigated by the oxidant pulsemethod. Upon injection of nitrogenous oxides to anaerobic cellsin darkness, an alkaline transient in the external medium wasobserved, followed by acidification. The alkaline transientwas enhanced by carbonyl cyanide m-chlorophenylhydrazone. When a viologen dye was used as an electron donor in the presenceof 1 mM Af-ethylmaleimide and 0.1 mM 2-n-heptyl-4-hydroxyquinoline-N-oxideto preclude respiration-linked H+ extrusion, addition of KNO3,KNO2 and N2O caused only a rapid alkalinization. The H+ consumptionstoichiometries, H+/2e ratios for NO3 reductionto NO2, NO2 reduction to 1/2 N2O and N2O reductionto N2 were 1.90, 3.18 and 2.04, respectively.These values agreed well with the fact that all reductions ofnitrogenous oxides in denitrification occur on the periplasmicside of the cytoplasmic membrane. When corrected for H+ consumption in the periplasm, the H+ extrusionstoichiometries, H+/2e ratios with endogenous substratesin the presence of K+/valinomycin for NO3 reduction toNO2, NO2 reduction to 1/2 N2O and N2O reductionto N2 were 4.05, 4.95 and 6.01, respectively. (Received August 4, 1982; Accepted January 13, 1983) |
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