Abstract
Growth and nitrogenase activity were studied in cultures ofAzotobacter vinelandii growing with dinitrogen, ammonium sulfate, aspartic acid or yeast extract. Nitrogenase activity was measured by means of the C2H2 reduction test.
In the presence of ammonium sulfate nitrogenase is completely repressed. After exhaustion of ammonia its activity is restored following a diauxic lag period of 30 min. With aspartic acid nitrogenase activity is partially repressed, and growth yield is higher than in the culture growing with N2 only. This is due to simultaneous use of dinitrogen and aspartate. Fluctuations of nitrogenase activity occurring during exponential growth and the mechanism of their regulation are discussed.
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Abbreviations
- NA:
-
nitrogenase activity
- BNF:
-
Burk's nitrogen free medium
References
Baak, J. M., Postma, P. W.: Oxidative phosphorylation in intactAzotobacter vinelandii. FEBS Letters19, 189–192 (1971)
Biggins, D. R., Postgate, J. R.: Nitrogen fixation by cultures and cell free extracts ofMycobacterium flavum. J. gen. Microbiol.56, 181–193 (1969)
Carnahan, J. E., Mortenson, L. E., Mower, H. F., Castle, J. E.: Nitrogen fixation in cell-free extracts ofClostridium pasteurianum. Biochim. biophys. Acta (Amst.)44, 520–535 (1960)
Daesch, G., Mortenson, L. E.: Effect of ammonia on the synthesis and function of the N2-fixing enzyme system inClostridium pasteurianum. J. Bact.110, 103–109 (1972)
Dalton, H., Mortenson, L. E.: Dinitrogen (N2) fixation (with a biochemical emphasis). Bact. Rev.36, 231–260 (1972)
Dilworth, M. J.: Acetylene reduction by nitrogen-fixing preparations fromClostridium pasteurianum. Biochim. biophys. Acta (Amst.)127, 285–294 (1966)
Drews, G.: Mikrobiologisches Praktikum für Naturwissenschaftler. Heidelberg-Berlin-New York: Springer 1968
Drozd, J. W., Postgate, J. R.: Interference by oxygen in the acetylene-reduction test for aerobic nitrogen-fixing bacteria. J. gen. Microbiol.60, 427–429 (1970)
Drozd, J. W., Tubb, R. S., Postgate, J. R.: A chemostat study of the effect of fixed nitrogen sources on nitrogen fixation, membranes and free amino acids inAzotobacter chroococcum. J. gen. Microbiol.73, 221–232 (1972)
Hardy, R. W. F., Holsten, R. D., Jackson, E. K., Burns, R. C.: The acetylene-ethylene assay for N2 fixation: Laboratory and field evaluation. Plant Physiol.43, 1185–1207 (1968)
Johnston, D. B., Fishbein, J. R.: Identification ofAzotobacter species by fluorescence and cell analysis. J. gen. Microbiol.14, 330 (1956)
Kelly, M.: Some properties of purified nitrogenase ofAzotobacter chroococcum. Biochim. biophys. Acta (Amst.)171, 9–22 (1969)
Kennedy, I. R., Morris, J. A., Mortenson, L. E.: N2 fixation by purified components of the N2-fixing system ofClostridium pasteurianum. Biochim. biophys. Acta (Amst.)153, 777–786 (1968)
Mahl, M. C., Wilson, P. W.: Nitrogen fixation by cell-free extracts ofKlebsiella pneumoniae. Canad. J. Microbiol.14, 33–38 (1968)
Moustafa, E., Mortenson, L. E.: Acetylene reduction by nitrogen fixing extracts ofClostridium pasteurianum: ATP requirement and inhibition by ADP. Nature (Lond.)216, 1241–1242 (1967)
Moustafa, E., Mortenson, L. E.: Properties of azoferredoxin purified from nitrogen-fixing extracts ofClostridium pasteurianum. Biochim. biophys. Acta (Amst.)172, 106–115 (1969)
Neilson, A., Rippka, R., Kunisawa, R.: Heterocyst formation and nitrogenase synthesis inAnabaena sp. A kinetic study. Arch. Mikrobiol.76, 139–150 (1971)
Newton, J. W., Wilson, P. W., Burris, R. H.: Direct demonstration of ammonia as an intermediate in nitrogen fixation byAzotobacter. J. biol. Chem.204, 445–453 (1953)
Ohmari, M., Hattori, A.: Effect of nitrite on nitrogen-fixation by the blue-green algaAnabaena cylindrica. Plant and Cell Physiol.13, 589–599 (1972)
Parejko, R. A., Wilson, P. W.: Regulation of nitrogenase synthesis byKlebsiella pneumoniae. Canad. J. Microbiol.16, 681–685 (1970)
Parker, C. A.: Effect of oxygen on the fixation of nitrogen byAzotobacter. Nature (Lond.)173, 780 (1954)
Patil, R. B., Pengra, R. M., Yoch, D. C.: Effects of nitrogen supplements on nitrogen fixation byAerobacter aerogenes. Biochim. biophys. Acta (Amst.)136, 1–5 (1967)
Pengra, R. M., Wilson, P. W.: Physiology of nitrogen fixation byAerobacter aerogenes. J. Bact.75, 21–25 (1958)
Postgate, J. R.: The acetylene reduction test for nitrogen fixation. In: Methods in microbiology, Vol. 6B, pp. 343–356, J. R. Norris and D. W. Ribbons, Eds. London-New York: Academic Press 1972
Schmidt-Lorenz, W., Rippel-Baldes, A.: Wirkung des Sauerstoff-Partialdruckes auf Wachstum und Stickstoffbindung vonAzotobacter chroococcum Beij. Arch. Mikrobiol.28, 45–68 (1957)
Shah, V. K., Davis, L. C., Brill, W. J.: Nitrogenase. I. Repression and derepression of the iron-molybdenum and iron proteins of nitrogenase inAzotobacter vinelandii. Biochim. biophys. Acta (Amst.)256, 498–511 (1972)
Wilson, P. W., Hull, J. F., Burris, R. H.: Competition between free and combined nitrogen in nutrition ofAzotobacter. Biochemistry29, 289–294 (1943)
Yoch, D. C., Pengra, R. M.: Effects of amino acids on the nitrogenase system ofKlebsiella pneumoniae. J. Bact.92, 618–622 (1966)
Zelitch, I.: Simultaneous use of molecular nitrogen and ammonia byClostridium pasteurianum. Proc. nat. Acad. Sci. (Wash.)37, 559–565 (1951)
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Gadkari, D., Stolp, H. Influence of nitrogen source on growth and nitrogenase activity inAzotobacter vinelandii . Arch. Microbiol. 96, 135–144 (1974). https://doi.org/10.1007/BF00590170
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DOI: https://doi.org/10.1007/BF00590170