Abstract
The primary steps of N2, ammonia and nitrate metabolism in Klebsiella pneumoniae grown in a continuous culture are regulated by the kind and supply of the nitrogenous compound. Cultures growing on N2 as the only nitrogen source have high activities of nitrogenase, unadenylated glutamine synthetase and glutamate synthase and low levels of glutamate dehydrogenase. If small amounts of ammonium salts are added continuously, initially only part of it is absorbed by the organisms. After 2–3 h complete absorption of ammonia against an ammonium gradient coinciding with an increased growth rate of the bacteria is observed. The change in the extracellular ammonium level is paralleled by the intracellular glutamine concentration which in turn regulates the glutamine synthetase activity. An increase in the degree of adenylation correlates with a repression of nitrogenase synthesis and an induction of glutamate dehydrogenase synthesis. Upon deadenylation these events are reversed.—After addition of nitrate ammonia appears in the medium, probably due to the action of a membrane bound dissimilatory nitrate reductase.—Addition of dinitrophenol causes transient leakage of intracellular ammonium into the medium.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bishop, P. E., McParland, R. H., Evans, H. J.: Inhibition of the adenylylation of glutamine synthetase by methionine sulfone during nitrogenase derepression. Biochem. biophys. Res. Commun. 67, 774–781 (1975)
Brenchley, J. E., Prival, M. J., Magasanik, B.: Regulation of the synthesis of enzymes responsible for glutamate formation in Klebsiella aerogenes. J. biol. Chem. 248, 6122–6128 (1973)
Cleland, W. W.: Steady state kinetics. Enzymes 2, 1–65 (1970)
Costerton, J. W., Ingram, J. M., Cheng, K.-J.: Structure and function of the cell envelope of gram-negative bacteria. Bact. Rev. 38, 87–110 (1974)
Foor, F., Jansen, K. A., Magasanik, B.: Regulation of synthesis of glutamine synthetase by adenylylated glutamine synthetase. Proc. nat. Acad. Sci. (Wash.) 72, 4844–4848 (1975)
Gancedo, C., Holzer, H.: Enzymatic inactivation of glutamine synthetase in enterobacteriaceae. Europ. J. Biochem. 4, 190–192 (1968)
Goa, J.: A microbiuret method for protein determination. Determination of tatal protein in cerebrospinal fluid. Scand. J. clin. Lab. Invest. 5, 218–222 (1953)
Grant, W. D., Sutherland, I. W., Wilkinson, J. F.: Exopolysaccharide colanic acid and its occurrence in the enterobacteriaceae. J. Bact. 100, 1187–1193 (1969)
Hardy, R. W. F., Knight, E., Jr.: Reductant-dependent adenosine triphosphatase of nitrogen-fixing extracts of Azotobacter vinelandii. biochim. biophys. Acta (Amst.) 122, 520–531 (1966)
Harold, F. M.: Conservation and transformation of energy by bacterial membranes. Bact. Rev. 36, 172–230 (1972)
Harold, F. M., Baarda, J. R.: Inhibition of membrane transport in Streptococcus faecalis by uncouplers of oxidative phosphorylation and its relationship to proton conduction. J. Bact. 96, 2025–2034 (1968)
Harold, F. M., Pavlasová, E., Baarda, J. R.: A transmembrane pH gradient in Streptococcus faecalis: Origin, and dissipation by proton conductors and N,N′-dicyclohexylcarbodiimide. Biochim. biophys. Acta (Amst.) 196, 235–244 (1970)
Holzer, H., Duntze, W.: Metabolic regulation by chemical modification of enzymes. Ann. Rev. Biochem. 40, 345–374 (1971)
Fleiner, D.: Quantitative relations for the repression of nitrogenase synthesis in Azotobacter vinelandii by ammonia. Arch. Microbiol. 101, 153–159 (1974)
Kleiner, D.: Ammonium uptake by nitrogen fixing bacteria. I. Azotobacter vinelandii. Arch. Microbiol. 104, 163–169 (1975)
Kováč, L., Kužela, Š.: Effect of uncopuling agents and azide on the synthesis of β-galactosidase in aerobically and anaerobically, grown Escherichia coli. Biochim. biophys. Acta (Amst.) 127, 355–365 (1966)
Magasanik, B., Prival, M. J., Brenchley, J. E., Tyler, B. M., De-Leo, A. B., Streicher, S. L., Bender, R. A., Paris, C. G.: Glutamine synthetase as a regulator of enzyme synthesis. Curr. Top. Cell. Reg. 8, 119–138 (1974)
Mahl, M. C., Wilson, P. W.: Nitrogen fixation by cell-free extracts from Klebsiella pneumoniae. Canad. J. Microbiol. 14, 33–38 (1968)
Marier, J. R., Boulet, M.: Direct determination of citric acid in milk with an improved pyridine-acetic anhydride method. J. Dairy Sci. 41, 1683–1692 (1958); ref. in Anal. Abstr. 6, 3762 (1959)
Meers, J. J., Tempest, D. W., Brown, C. M.: “Glutamine(amide): 2-oxoglutarate amino transferase oxido-reductase (NADP)”, an enzyme involved in the synthesis of glutamate by some bacteria. J. gen. Microbiol. 64, 187–194 (1970)
Mickelson, M. N.: Effect of uncoupling agents and respiratory inhibitors on the growth of Streptococcus agalactiae. J. Bact. 120, 733–740 (1974)
Nagatani, H., Shimizu, M., Valentine, R. C.: The mechanism of ammonia assimilation in nitrogen fixing bacteria. Arch. Microbiol. 79, 164–175 (1971)
Pareijko, R. A., Wilson, P. W.: Regulation of nitrogenase synthesis by Klebsiella pneumoniae. Canad. J. Microbiol. 16, 681–685 (1970)
Pavlasova, E., Harold, F. M.: Energy coupling in the transport of β-galactoside by Escherichia coli: Effect of proton conductors. J. Bact. 98, 198–204 (1969)
Pengra, R. M., Wilson, P. W.: Physiology of nitrogen fixation by Aerobacter aerogenes J. Bact. 75, 21–25 (1958)
Riebeling, V., Thauer, R. K., Jungermann, K.: The internal-alkaline pH gradient, sensitive to uncoupler and ATPase inhibitor, in growing Clostridium pasteurianum. Europ. J. Biochem. 55, 445–453 (1975)
Schutt, H., Holzer, H.: Biological function of the ammonia-induced inactivation of glutamine synthetase in Escherichia coli. Europ. J. Biochem. 26, 68–72 (1972)
Shanmugam, K. T., Chan, I., Morandi, C.: Regulation of nitrogen fixation. Nitrogenase-derepressed mutants of Klebsiella pneumoniae. Biochim. biophys. Acta (Amst.) 408, 101–111 (1975)
Shapiro, B. M., Stadtman, E. R.: Glutamine synthetase (Escherichia coli). Meth. Enzymol. 17, 910–922 (1972)
Stadtman, E. R., Ginsburg, A., Ciardi, J. E., Yeh, J., Hennig, S. B., Shapiro, B. M.: Multiple molecular forms of glutamine synthetase produced by enzyme catalyzed adenylylation and deadenylylation reactions. Advanc. Enzyme Reg. 8, 99–118 (1970)
Streicher, S. L., Shanmugam, K. T., Ausubel, F., Morandi, C., Goldberg, R. B.: Regulation of nitrogen fixation in Klebsiella pneumoniae: Evidence for glutamine synthetase as a regulator of nitrogenase synthesis. J. Bact. 120, 815–821 (1974)
Tubb, R. S.: Glutamine synthetase and ammonium regulation of nitrogenase synthesis in Klebsiella. Nature (Lond.) 251, 481–485 (1974)
Tubb, R. S., Postgate, J. R.: Control of nitrogenase synthesis in Klebsiella pneumoniae. J. gen. Microbiol. 79, 103–117 (1973)
Van't Riet, J., Planta, R. J.: Purification, structure and properties of the respiratory nitrate reductase of Klebsiella aerogenes. Biochim. biophys. Acta (Amst.) 379, 81–94 (1975)
Yoch, D. C., Pengra, R. M.: Effect of amino acids on the nitrogenase system of Klebsiella pneumoniae. J. Bact. 92, 618–622 (1966)
Zumft, W. G., Mortenson, L. E.: The nitrogen-fixing complex of bacteria. Biochim. biophys. Acta (Amst.) 416, 1–52 (1975)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kleiner, D. Ammonium uptake and metabolism by nitrogen fixing bacteria. Arch. Microbiol. 111, 85–91 (1976). https://doi.org/10.1007/BF00446553
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00446553