Abstract
In order to examine the mediatory role of proton motive force (∆p) or proton ATPase in H2 production by Rhodobacter sphaeroides, ∆p was determined under anaerobic conditions in the dark, and the ATPase activity has been studied in R. sphaeroides strain A-10, isolated from Arzni mineral springs in Armenia. Membrane potential (∆φ) was measured from the distribution of tetraphenylphosphonium cation; pH gradient (∆pH) was the difference between the external and cytoplasmic pH values, and the latter was measured by 9-aminoacridine (9-AA) fluorescence changes. At pH 7.5, ∆φ was of −94 mV and the reversed ∆pH was +30 mV, resulting in ∆p of −64 mV. The addition of N,N′-dicyclohexylcarbodiimide (DCCD), the F0F1–ATPase inhibitor, was not affect ∆φ. It was shown that ∆φ varies nearly linearly with ΔpH, ∆φ increased from −57.1 mV at pH 6.0 to −103.8 mV at pH 8.0; it was compensated at high external pH by a reversed ∆pH, resulting in a low ∆p under anaerobic-dark conditions. Intracellular ATP concentrations and energetic charge (EC) were measured to evaluate a metabolism activity of R. sphaeroides.
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Acknowledgments
The author thank Prof. P. Kazaryan (Eolyan Hematology Center, Yerevan) for his help in ANP determination and valuable comments. This study was supported by the Armenian National Science and Education Fund, USA (ANSEF NS-Biotechnology-1668) and by the Research Grant from the Ministry of Education and Science of Armenia (#1012-2008).
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Hakobyan, L., Gabrielyan, L. & Trchounian, A. Proton Motive Force in Rhodobacter sphaeroides Under Anaerobic Conditions in the Dark. Curr Microbiol 62, 415–419 (2011). https://doi.org/10.1007/s00284-010-9723-1
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DOI: https://doi.org/10.1007/s00284-010-9723-1