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Hydrogen oxidation and efficiency of nitrogen fixation in succinate-limited chemostat cultures ofRhizobium ORS 571

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Abstract

Rhizobium ORS 571, isolated from stem nodules of the tropical legumeSesbania rostrata is able to grow in the chemostat with molecular nitrogen as sole nitrogen source at a specific growth rate of 0.1 h-1. Samples from nitrogenfixing cultures showed high acetylene reduction activities: 1,500 nmol ethylene formed per milligram dry weight per hour. Under nitrogen-fixing conditions an uptake hydrogenase is induced. Ammonia-assimilating cultures, without additional hydrogen, did not induce hydrogenase. The addition of hydrogen to succinate-limited nitrogen-fixing cultures resulted in an increase in the molar growth yield on succinate (Y succinate) from 27 to 35 and a slight decrease in the molar growth yield on oxygen (\(Y_{O_2 }\)), showing that hydrogen oxidation is less energy-yielding than the oxidation of endogenous substrates. Respiration-driven proton translocation measured with starved cells indicated the functioning of site 1 and 2 of oxidative phosphorylation. Cytochrome spectra showed that cytochromea 600, present at high dissolved oxygen tension (d.o.t.) almost completely disappeared at low d.o.t. In flash-photolysis spectra only thea-type cytochrome could be detected as an oxidase in cells both grown at high and low d.o.t. Growth yields in ammonia-assimilating cultures were higher than those measured in nitrogen-fixing cultures. Assuming two sites of oxidative phosphorylation, a molar growth yield on ATP (Y ATP) of about 3 and 6 was calculated for respecticely nitrogen-fixing and ammonia-assimilating cultures. TheY ATP under nitrogen-fixing conditions is dependent on the amount of H2 formed per mol N2 fixed (H2/N2 ratio). A method has been described to calculate the total amount of ATP use by nitrogenase during the fixation of 1 mol N2 (ATP/N2 ratio) and H2/N2 ratios in aerobic nitrogen fixing organisms. This calculation yielded that nitrogen fixation inRhizobium ORS 571 is a high ATP-consuming process. The calculated ATP/N2 and H2/N2 ratios were respectively 42 and 7.5.

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Abbreviations

d.o.t.:

dissolved oxygen tension

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Authors and Affiliations

  1. Department of Microbiology, Biological Laboratory, Vrije Universiteit, Postbus 7161, NL-1007 MC, Amsterdam, The Netherlands

    Hein Stam, Henk W. van Verseveld, Wytske de Vries & Adriaan H. Stouthamer

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  1. Hein Stam
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  2. Henk W. van Verseveld
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  3. Wytske de Vries
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  4. Adriaan H. Stouthamer
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Stam, H., van Verseveld, H.W., de Vries, W. et al. Hydrogen oxidation and efficiency of nitrogen fixation in succinate-limited chemostat cultures ofRhizobium ORS 571. Arch. Microbiol. 139, 53–60 (1984). https://doi.org/10.1007/BF00692712

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  • DOI: https://doi.org/10.1007/BF00692712

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