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‘Respiratory protection’ of the nitrogenase in dinitrogen-fixing cyanobacteria

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Abstract

Several filamentous and unicellular cyanobacteria were grown photoautotrophically with nitrate or dinitrogen as N-sources, and some respiratory properties of the cells or isolated plasma (CM) and thylakoid (ICM) membranes were compared. Specific cytochrome c oxidase activities in membranes from dinitrogen-fixing cells were between 10- and 50-times higher than those in membranes from nitrate-grown cells, ICM of heterocysts but CM of unicells being mainly responsible for the stimulation. Whole cell respiration (oxygen uptake) of diazotrophic unicells paralleled increased cytochrome oxidase activities of the isolated membranes. Mass spectrometric measurements of the uptake of isotopically labeled oxygen revealed that (low) light inhibited respiration of diazotrophic unicells to a much lesser degree than that of nitrate-grown cells which indicates the prevailing (respiratory) role of CM in the former. Normalized growth yields of diazotrophic unicells grown in continuous light were significantly higher than those of cells grown in a 12/12 hrs light/dark cycle. Mass spectrometry showed that overall nitrogen uptake by the former was higher than by the latter; in particular, and in marked contrast to the time course of nitrogenase activity (acetylene reduction) there was no appreciable nitrogen uptake or protein synthesis during dark periods; likewise, there was no 14-CO2 fixation, nor chloropholl synthesis, nor cell division in the dark. By contrast, growth in continuous light gave sustained rates of nitrogen and carbon dioxide incorporation over the whole time range. Our results will be discussed in terms of ‘respiratory protection’ as an essential strategy of keeping apart nitrogenase and oxygen, either atmospheric or photosynthetically produced within the same cell.

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Abbreviations

CM:

cytoplasmic or plasma membrane

ICM:

intracytoplasmic or thylakoid membrane

CCCP:

carbonyl cyanide m-chlorophenylhydrazone

PCC:

Pasteur Culture Collection

References

  • Almon H and Böger P 1988 Nitrogen and hydrogen metabolism: Induction and measurement. Methods Enzymol. 167, 459–467.

    Google Scholar 

  • Bradford M M 1976 A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254.

    PubMed  Google Scholar 

  • Carpenter E J and Price C C 1976 MarineOscillatoria (Trichodesmium): Explanation for aerobic nitrogen fixation without heterocysts. Science 191, 1278–1280.

    PubMed  Google Scholar 

  • Dalton H and Postgate J R 1969 Effect of oxygen on growth ofAzotobacter chroococcum in batch and continuous cultures. J. Gen. Microbiol. 54, 463–473.

    Google Scholar 

  • Dimon B, Gans P and Peltier G 1988 Mass spectrometric measurement of photosynthetic and respiratory oxygen exchange. Methods Enzymol. 167, 686–691.

    Google Scholar 

  • Erber W W A, Nitschmann W H, Muchl R and Peschek G A 1986 Endogenous energy supply to the plasma membrane of dark aerobic cyanobacteriumAnacystis nidulans: ATPase-independent efflux of H+ and Na+ from respiring cells. Arch. Biochem. Biophys. 247, 28–39.

    PubMed  Google Scholar 

  • Ernst A, Kirschenlohr H, Diez J and Böger P 1984 Glycogen content and nitrogenase activity inAnabaena variabilis. Arch. Microbiol. 140, 120–125.

    Google Scholar 

  • Fry I V, Huflejt M, Erber W W A, Peschek G A and Packer L 1986 The role of respiration during adaptation of the freshwater cyanobacterium Synechococcus 6311 to salinity. Arch. Biochem. Biophys. 244, 686–691.

    PubMed  Google Scholar 

  • Gallon J R and Chaplin A E 1988 Nitrogen fixation.In Biochemistry of the Algae and Cyanobacteria. Eds. L JRogers and J RGallon. pp 147–173. Oxford Science Publication, Clarendon Press. Oxford.

    Google Scholar 

  • Haselkorn R 1978 Heterocysts. Annu. Rev. Plant Physiol. 29, 319–344.

    Google Scholar 

  • Hoch G, Owens O H and Kok B 1963 Photosynthesis and respiration. Arch. Biochem. Biophys. 101, 171–180.

    PubMed  Google Scholar 

  • Jensen B B and Cox R P 1988 Measurement of hydrogen exchange and nitrogen uptake by mass spectrometry. Methods Enzymol. 167, 467–474.

    Google Scholar 

  • Jones C W, Brice J M, Wright V and Ackrell B A C 1973 Respiratory protection of nitrogenase inAzotobacter vinelandii. FEBS Lett. 29, 77–81.

    PubMed  Google Scholar 

  • Kallas T, Rippka R, Coursin T, Revière M-C, Tandeaude Marsac N and Cohen-Bazire G 1983 Aerobic nitrogen fixation by nonheterocystous cyanobacteria.In Photosynthetic Prokaryotes—Cell Differentiation and Function. Eds. G CPapageorgiou and LPacker. pp 281–302, Elsevier Biomedical Publishers, New York.

    Google Scholar 

  • Lung N J and Fay P 1971 The heterocysts of blue-green algae. II. Details of ultrastructure. Proc. Roy. Soc. Lond. B. 178, 193–203.

    Google Scholar 

  • Mackinney G 1941 Absorption of light by chlorophyll solutions. J. Biol. Chem. 140, 315–322.

    Google Scholar 

  • Mitsui A, Kumazawa S, Takahashi A, Ikemoto H, Cao S and Arai T 1986 Strategy by which nitrogen-fixing unicellular cyanobacteria grow photoautotrophically. Nature 323, 720–722.

    Google Scholar 

  • Molitor V and Peschek G A 1986 Respiratory eletron transport in plasma and thylakoid membrane preparations from the cyanobacteriumAnacystis nidulans. FEBS Lett. 195, 145–150.

    Google Scholar 

  • Mortenson L E and Thorneley R N F 1979 Structure and function of nitrogenase. Annu. Rev. Biochem. 48, 387–418.

    PubMed  Google Scholar 

  • Murata N and Omata T 1988 Isolation of cyanobacterial plasma membranes. Methods Enzymol 167, 245–251.

    Google Scholar 

  • Pearson H W and Howsley R 1980 Concomitant photo-autotrophic growth and nitrogenase activity by cyanobacteriumPlectonema boryanum in continuous culture. Nature 288, 263–265.

    Google Scholar 

  • Pearson H W, Howsley R, Kjeldsen C K and Walsby A E 1979 Aerobic nitrogenase activity associated with a non-heterocystous filamentous cyanobacterium. FEMS Microbiol. Lett. 5, 163–167.

    Google Scholar 

  • Peschek G A (1987) Respiratory electron transport.In The Cyanobacteria. Eds. PFay and CVanBaalen. pp 119–161. Elsevier Biomedical Publishers. Amsterdam.

    Google Scholar 

  • Peschek G A, Wastyn M, Trnka M, Molitor V, Fry I V and Packer L 1989 Characterization of the cytochrome c oxidase in isolated and purified plasma membranes from the cyanobacteriumAnacystis nidulans. Biochemistry 28, 3057–3063.

    PubMed  Google Scholar 

  • Peschek G A, Molitor V, Trnka M, Wastyn M and Erber W 1988 Characterization of cytochrome-c oxidase in isolated and purified plasma and thylakoid membranes from cyanobacteria. Methods Enzymol. 167, 437–449.

    Google Scholar 

  • Peschek G A, Hinterstoisser B, Wastyn M, Kuntner O, Pineau B, Missbichler A and Lang J 1989 Chlorophyll precursors in the plasma membrane of a cyanobacterium,Anacystis nidulans. J. Biol. Chem. 264, 11827–11832.

    PubMed  Google Scholar 

  • Radmer R and Ollinger O 1980 Measurement of the oxygen cycle: the mass spectrometric analysis of gases dissolved in a liquid phase. Methods Enzymol. 69, 547–560.

    Google Scholar 

  • Robson R L and Postgate J R 1980 Oxygen and hydrogen in biological nitrogen fixation. Annu. Rev. Microbiol. 34, 183–207.

    PubMed  Google Scholar 

  • Stal L J and Krumbein W E 1985 Oxygen protection of nitrogenase in the aerobically nitrogen-fixing, non-heterocystous cyanobacteriumOscillatoria sp. Arch. Microbiol. 143, 72–76.

    Google Scholar 

  • Stanier R Y and Cohen-Bazire G 1977 Phototrophic prokaryotes: The cyanobacteria. Annu. Rev. Microbiol. 31, 225–274.

    PubMed  Google Scholar 

  • Stewart W D P 1980 Some aspects of structure and function in N2-fixing cyanobacteria. Annu. Rev. Microbiol. 34, 497–536.

    PubMed  Google Scholar 

  • Wastyn M, Achatz A, Molitor V and Peschek G A 1988 Respiratory activities and aa3-type cytochrome oxidase in plasma and thylakoid membranes from vegetative cells and heterocysts of the cyanobacterium Anabaena ATCC 29413. Biochim. Biophys. Acta 935, 217–224.

    Google Scholar 

  • Winkenbach F and Wolk C P 1973 Activities of enzymes of the oxidative and the reductive pentose phosphate pathways in heterocysts of a blue-green alga. Plant Physiol. 52, 480–484.

    Google Scholar 

  • Yates M G and Jones C W 1974 Respiration and nitrogen-fixation in Azotobacter. Adv. Microb. Physiol. 11, 97–135.

    Google Scholar 

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Peschek, G.A., Villgrater, K. & Wastyn, M. ‘Respiratory protection’ of the nitrogenase in dinitrogen-fixing cyanobacteria. Plant Soil 137, 17–24 (1991). https://doi.org/10.1007/BF02187427

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