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Membrane-bound NADPH dehydrogenase- and ferredoxin: NADP oxidoreductase activity involved in electron transport during acetate oxidation to CO2 in Desulfobacter postgatei

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Abstract

Desulfobacter postgatei grows on acetate and sulfate as energy source. The oxidation of acetate to 2 CO2 proceeds via the citric acid cycle involving membrane-bound succinate dehydrogenase and membrane-bound malate dehydrogenase. We report here that the organism contains membrane-bound NADPH dehydrogenase and ferredoxin: NADP oxidoreductase for the reoxidation of NADPH and reduced ferredoxin generated during isocitrate- and 2-oxoglutarate oxidation, respectively. The presence of proton translocating ATPase activity is also described.

NADPH dehydrogenase and succinate dehydrogenase were found to be electrically connected within the membrane and electron transfer between these two enzymes was shown to be coupled with proton translocation. The membrane fraction catalyzed the oxidation of NADPH with fumarate and the reduction of NADP with succinate. NADPH oxidation with fumarate was stimulated by protonophores and inhibited by the proton translocating ATPase inhibitor dicyclohexylcarbodiimide (DCCD) and by heptylhydroxyquinoline-N-oxide (HQNO); inhibition by DCCD was relieved by protonophores. NADP reduction with succinate was dependent on ATP and inhibited by protonophores, DCCD, and HQNO. The membrane fraction also mediated the oxidation of NADPH with the water soluble menaquinone analogue dimethylnaphthoquinone (DMN) and the reduction of fumarate with DMNH2. Only the former reaction was stimulated by protonophores and only the latter reaction was inhibited by HQNO. This suggests that the NADPH dehydrogenase reaction is the site of energy conservation and the succinate dehydrogenase is the site of HQNO inhibition.

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Abbreviations

APS:

Adenosine 5′-phosphosulfate

DCCD:

N,N′-dicyclohexylcarbodiimide

DCPIP:

2,6-dichloroindophenol

DMN:

2,3-dimethyl-1,4-naphthoquinone

DTT:

DL-1,4-dithiothreitol

HQNO:

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

TCS:

3,5,3′,4′-tetrachlorosalicylanilide

Tricine:

N-tris-(hydroxymethyl)methylglycine

TTFB:

4,5,6,7-tetrachloro-2′-trifluoromethylbenzimidazole

SF-6847:

3,5-di-tert-butyl-4-hydroxybenzylidenemalononitrile

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  1. Mikrobiologie, Fachbereich Biologie, Philipps-Universität, Karl-von-Frisch-Straße, D-3550, Marburg, Germany

    Dieter Möller-Zinkhan & Rudolf K. Thauer

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  1. Dieter Möller-Zinkhan
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Möller-Zinkhan, D., Thauer, R.K. Membrane-bound NADPH dehydrogenase- and ferredoxin: NADP oxidoreductase activity involved in electron transport during acetate oxidation to CO2 in Desulfobacter postgatei . Arch. Microbiol. 150, 145–154 (1988). https://doi.org/10.1007/BF00425154

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