Summary
The free amino acid pools in the nitrogen-fixing blue-green algae Anabaena cylindrica, A. flos-aquae and Westiellopsis prolifica contain a variety of amino acids with aspartic acid, glutamic acid and the amide glutamine being present in much higher concentrations than the others. This pattern is characteristic of that found in organisms having glutamine synthetage/glutamate synthetase [glutamine amide-2-oxoglutarate amino transferase (oxido-reductase)] as an important pathway of ammonia incorporation. Under nitrogen-starved conditions the level of acetylene reduction (nitrogen fixation) and the glutamine pool both increase but the free ammonia pool decreases, suggesting that ammonia rather than glutamine regulates nitrogen fixation.
Glutamine synthetase has been demonstrated in Anabaena cylindrica using the γ-glutamyl transferase assay and also using a biosynthetic assay in which Pi release from ATP during glutamine synthesis was measured. The enzyme (γ-glutamyl transferase assay) is present in nitrogen-fixing cultures and activity is higher in aerobic than in microaerophilic cultures. Ammonium-grown cultures have lowest levels of all and activity in the presence of nitrate-nitrogen (150 mg nitrogen 1-1) is lower than in aerobic cultures growing on elemental nitrogen. Ammonium-nitrogen and nitrate-nitrogen have no effect on glutamine synthetase in vitro. Glutamate synthetase also operates in nitrogen-fixing cultures of Anabaena cylindrica.
Similar content being viewed by others
References
Allen, M. B., Arnon, D. I.: Studies on nitrogen-fixing blue-green algae. I: Growth and nitrogen fixation by Anabaena cylindrica Lemm. Plant Physiol. 30, 366–372 (1955).
Benson, J. V., Jr., Gordon, M. F., Patterson, J. A.: Accelerated chromatographic analysis of amino acids. Analyt. Biochem. 18, 228–240 (1967).
Bothe, H.: Photosynthetische Stickstoffixierung mit einem zellfreien Extrakt aus der Blaualge Anabaena variabilis. Ber. dtsch. bot. Ges. 83, 421–432 (1970).
Brown, C. M., Stanley, S. O.: Environment-mediated changes in the cellular content of the “pool” constituents and their associated changes in cell physiology. J. appl. Chem. Biotechnol. 22, 363–389 (1972).
Burris, R. H., Wilson, P. W.: Methods for measurement of nitrogen fixation. In: Methods in enzymology (eds. S. Colowick and N. O. Kaplan) vol. 4, p. 355–365 (1957).
Dharmawardene, N., Stewart, W. D. P.: Combined nitrogen and nitrogen fixation in the blue-green alga Anabaena cylindrica (in press).
Fogg, G. E.: Growth and heterocyst production in Anabaena cylindrica Lemm. II. In relation to carbon and nitrogen metabolism. Ann. Bot. (Lond.), N.S. 13, 241–259 (1949).
Hoare, D. S., Hoare, S. L., Moore, R. B.: The photoassimilation of organic compounds by autotrophic blue-green algae. J. gen. Microbiol. 49, 351–370 (1967).
Holm-Hansen, O., Brown, G. W.: Ornithine cycle enzymes in the blue-green alga Nostoc muscorum. Plant Cell Physiol. Tokyo 4, 299–306 (1963).
Holzer, H.: Regulation of enzymes by enzyme catalyzed chemical modification. Advanc. Enzymol. 32, 297–326 (1969).
Hood, W., Carr, N. G.: Apparent lack of control by repression of arginine metabolism in blue-green algae. J. Bact. 107, 365–367 (1971).
Linko, P., Holm-Hansen, O., Bassham, J. A., Calvin, M.: Formation of radio-active citrulline during photosynthetic C14O2 fixation by blue-green algae. J. exp. Bot. 8, 147–156 (1964).
Mecke, D., Holzer, H.: Repression und Inaktivierung von Glutaminskythetase in E. coli durch NH +4 . Biochim. biophys. Acta (Amst.) 122, 341–351 (1966).
Meers, J. L., Pedersen, L.: Nitrogen assimilation by Bacillus licheniformis organisms growing in chemostat cultures. J. gen. Microbiol. 70, 277–286 (1972).
Meers, J. L., Tempest, D. W., Brown, C. M. “Glutamine (amide): 2-oxoglutarate aminotransferase oxido-reductase (NADP)”, an enzyme involved in the synthesis of glutamate by some bacteria. J. gen. Microbiol. 64, 187–194 (1970)
Nagatani, H., Schimizu, M., Valentine, R. C.: The mechanism of ammonia assimilation in nitrogen-fixing bacteria. Arch. Mikrobiol. 79, 164–175 (1971).
Parejko, R. A., Wilson, P. W.: Regulation of nitrogenase synthesis by Klebsiella pneumoniae. Canad. J. Microbiol. 16, 681–685 (1970).
Pate, J. S., Wallace, W.: Movement of assimilated nitrogen from the root system of the field pea (Pisum arvense L.). Ann. Bot. (Lond.), N.S. 28, 83–99 (1964).
Shapiro, B. M., Stadtman, E. R.: Glutamine synthetase (Escherichia coli), transferase assay, γ-glutamyl hydroxamate measured. In: Methods in Enzymology, vol. 17 A, p. 910–922 (1970).
Spackman, D. H., Stein, W. H., Moore, S.: Automatic recording apparatus for use in the chromatography of amino acids. Analyt. Chem. 30, 1190–1206 (1958).
Stanley, P. E.: The growth of Torulopsis utilis in aerated batch and continuous culture. Ph.D. thesis, University of Bristol (1964).
Stewart, W. D. P.: Biological and ecological aspects of nitrogen fixation by free-living micro-organisms. Proc. roy. Soc. B 172, 367–388 (1969).
Stewart, W. D. P.: Algal fixation of atmospheric nitrogen. Plant and Soil 32, 555–588 (1970).
Stewart, W. D. P.: Physiological studies on nitrogen-fixing blue-green algae. Plant and Soil Special volume 377–391 (1971).
Stewart, W. D. P., Fitzgerald, G. P., Burris, R. H.: In situ studies on N2 fixation using the acetylene reduction technique. Proc. nat. Acad. Sci. (Wash.) 58, 2071–2078 (1967).
Stewart, W. D. P., Mague, T., Fitzgerald, G. P., Burris, R. H.: Nitrogenase activity in Wisconsin lakes of differing degrees of eutrophication. New Phytologist 70, 497–509 (1971).
Tempest, D. W., Meers, J. L., Brown, C. M.: Synthesis of glutamate in Aerobacter aerogenes by a hitherto unknown route. Biochem. J. 117, 405–407 (1970).
Woolfolk, C. A., Shapiro, B., Stadtman, E. R.: Regulations of glutamine synthetase. 1. Purification and properties of glutamine synthetase from Escherichia coli. Arch. Biochem. 116, 177–192 (1966).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Dharmawardene, M.W.N., Stewart, W.D.P. & Stanley, S.O. Nitrogenase activity, amino acid pool patterns and amination in blue-green algae. Planta 108, 133–145 (1972). https://doi.org/10.1007/BF00386075
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00386075