1887

Abstract

SUMMARY: Increasing concentrations of ammonium ions in the medium of nitrogen-fixing, sulphate-limited continuous cultures of caused a proportionate repression of nitrogenase activity; free NH could be detected in the extracellular culture fluid only when nitrogenase activity was wholly repressed. The NH concentrations giving 50% or 100% repression were proportional to the population density. Nitrate ions repressed with similar stoichiometry; glutamate, glutamine and aspartate did not repress and were not metabolized; repressed and derepressed populations contained equal amounts and proportions of glutamate-forming enzymes. Repressed populations lacked both enzymatic components of nitrogenase. The intracellular free amino acid pools were typical of Gram-negative bacteria; an increase in the degree of repression was associated with an increase in the pool levels of ammonia, aspartate and glutamate. Nitrogen-fixing populations possessed a convoluted intracytoplasmic membrane system which was absent from ammonia-assimilating organisms, but the phospholipid contents of the two types of population were similar. All members of a half-repressed population possessed these membranes, but to a lesser extent that fully derepressed populations.

When N-fixing chemostat populations were abruptly exposed to repressive concentrations of ammonium succinate. repression occurred exponentially and nitrogenase activity disappeared from the culture faster than wash-out of stable enzyme. Repression was not alleviated by exogenous cyclic AMP. Derepression was complete, according to the acetylene test, within half a doubling time of disappearance of free ammonium ions from the culture.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-73-2-221
1972-11-01
2024-04-24
Loading full text...

Full text loading...

/deliver/fulltext/micro/73/2/mic-73-2-221.html?itemId=/content/journal/micro/10.1099/00221287-73-2-221&mimeType=html&fmt=ahah

References

  1. Baker K. 1968; Low cost continuous culture apparatus. Laboratory Practice 17:817–824
    [Google Scholar]
  2. Bartlett G. R. 1959; Phosphorus assay in column chromatography. Journal of Biological Chemistry 234:466–468
    [Google Scholar]
  3. Boltz D. F. 1958 Colorimetric Determination of Non-Metals. New York: Interscience Inc.;
    [Google Scholar]
  4. Bone D. H. 1971; Nitrogenase activity and nitrogen assimilation in Anabena flos-aquae growing in continuous culture. Archiv für Mikrobiologie 80:234–241
    [Google Scholar]
  5. Daesch G., Mortenson L. E. 1968; Sucrose catabolism in Clostridium pasteurianum and its relation to N2 fixation. Journal of Bacteriology 96:346–351
    [Google Scholar]
  6. Dalton H., Postgate J. R. 1969a; Effect of oxygen on growth of Azotobacter chroococcum in batch and continuous cultures. Journal of General Microbiology 54:463–473
    [Google Scholar]
  7. Dalton H., Postgate J. R. 1969b; Growth and physiology of Azotobacter chroococcum in continuous culture. Journal of General Microbiology 56:307–319
    [Google Scholar]
  8. Davis L. C., Shah V. K., Brill W. J., Orme- Johnson W. H. 1972; Nitrogenase. II. Changes in the EPR signal of component 1 (iron-molybdenum protein) of Azotobacter vinelandii during repression and derepression. Biochimica et biophysica acta 256,512–523
    [Google Scholar]
  9. Drozd J. W., Postgate J. R. 1970; Effects of oxygen on acetylene reduction, cytochrome content and respiratory activity of Azotobacter chroococcum . Journal of General Microbiology 63:63–73
    [Google Scholar]
  10. Folch J., Lees M., Sloane-Stanley G. H. 1957; A simple method for the isolation and purification of total lipids from animal tissues. Journal of Biological Chemistry 226:497–506
    [Google Scholar]
  11. Goerz R. D., Pengra R. M. 1961; Physiology of nitrogen function by a species of Achromobacter . Journal of Bacteriology 81:568–572
    [Google Scholar]
  12. Gorini L., Maas W. K. 1957; The potential for the formation of a biosynthetic enzyme in Escherichia coli . Biochimica et biophysica acta 25:208–209
    [Google Scholar]
  13. Hardy R. W. F., Holsten R. D., Jackson E. K., Burns R. C. 1968; The acetylene-ethylene assay for N2 fixation — laboratory and field evaluation. Plant Physiology 43:1185–1207
    [Google Scholar]
  14. Hill S., Drozd J. W., Postgate J. R. 1972; Environmental effects on the growth of nitrogen-fixing bacteria. Journal of Applied Chemistry and Biotechnology 22:541–558
    [Google Scholar]
  15. Kellenberger E., Ryter A., Sechaud J. 1958; Electron microscope study of DNA-containing plasms. II. Vegetative and mature phase DNA as compared with normal bacterial nucleoids in different physiological states. Journal of Biophysical and Biochemical Cytology 4:671–676
    [Google Scholar]
  16. Kelly M. 1969; The properties of purified nitrogenase of Azotobacter chroococcum . Biochimica et biophysica acta 171:9–22
    [Google Scholar]
  17. Kelly M., Klucas R. V., Burris R. H. 1967; Fractionation and storage of nitrogenase from Azotobacter vinelandii . Biochemical Journal 105:3C–5C
    [Google Scholar]
  18. Lascelles J., Szilagyi J. R. 1965; Phospholipid synthesis by Rhodopseudomonas spheriodes in relation to the formation of photosynthetic pigments. Journal of General Microbiology 38:55–64
    [Google Scholar]
  19. Luft J. M. 1961; Improvements in epoxy resin embedding methods. Journal of Biophysical and Biochemical Cytology 9:409–414
    [Google Scholar]
  20. Macura J. 1966; Continuous flow methods in soil microbiology. In Theoretical and Methodological Basis of Continuous Culture of Microorganisms, pp. 470–472 Edited by Malek I., Feud Z. Prague: Czechoslovak Academy of Sciences;
    [Google Scholar]
  21. Mahl M. C., Wilson P. W. 1967; Nitrogen fixation by cell-free extracts of Klebsiella pneumoniae . Canadian Journal of Microbiology 14:33–38
    [Google Scholar]
  22. Meers J. L., Tempest D. W., Brown C. M. 1970; ‘Glutamine (amide): 2-oxoglutarate amino transferase oxido-reductase (NADP)’, an enzyme involved in the synthesis of glutamate by some bacteria. Journal of General Microbiology 64:187–194
    [Google Scholar]
  23. Munson T. O., Burris R. H. 1969; Nitrogen fixation by Rhodospirillum rubrum grown in nitrogen- limited continuous culture. Journal of Bacteriology 97:1093–1098
    [Google Scholar]
  24. Nagatani H., Shimazu M., Valentine R. C. 1971; The mechanism of ammonia assimilation in nitrogen-fixing bacteria. Archiv für Mikrobiologie 79:164–175
    [Google Scholar]
  25. Neilson A., Rippka R., Kunisawa R. 1971; Heterocyst formation and nitrogenase synthesis in Anabena sp. - a kinetic study. Archiv für Mikrobiologie 76:139–150
    [Google Scholar]
  26. Oppenheim J., Marcus L. 1970a; Correlation of ultra-structure in Azotobacter vinelandii with nitrogen source for growth. Journal of Bacteriology 101:286–291
    [Google Scholar]
  27. Oppenheim J., Marcus L. 1970b; Induction and repression of nitrogenase and internal membranes in Azotobacter vinelandii . Bacteriological Proceedings,148
    [Google Scholar]
  28. Pangborn J., Marr A. G., Robrish S. A. 1962; Localisation of respiratory enzymes in intracytoplasmic membranes of Azotobacter agilis . Journal of Bacteriology 84:669–678
    [Google Scholar]
  29. Parejko R. A., Wilson P. W. 1970; Regulation of nitrogenase synthesis by Klebsiella pneumoniae . Canadian Journal of Microbiology 16:681–685
    [Google Scholar]
  30. Postgate J. R., Hunter J. R. 1962; The survival of starved bacteria. Journal of General Microbiology 29:233–263 Addendum, 1963 Journal of General Microbiology 34, 473
    [Google Scholar]
  31. Reynolds E. S. 1963; The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. Journal of Cell Biology 17:208–212
    [Google Scholar]
  32. Roberts R. B., Abelson P. H., Cowrie D. B., Bolton E. T., Britten R. J. 1955 Studies on Bio-synthesis in Escherichia coli. Publication 607. Washington D.C.: Carnegie Institution;
    [Google Scholar]
  33. St John R. T., Brill W. S. 1972; Inhibitory effect of methylalanine on glucose-grown Azotobacter vinelandii . Biochimica et biophysica acta 261:63–69
    [Google Scholar]
  34. Shah V. K., Davis L. C., Brill W. J. 1972; Nitrogenase. I. Repression and derepression of the iron- molybdenum and iron proteins in Azotobacter vinelandii . Biochimica et biophysica acta 256:498–511
    [Google Scholar]
  35. Smith R. V., Evans M. C. W. 1970; Soluble nitrogenase from vegetative cells of the blue-green alga Anabaena cylindrica . Nature, London 225:1253–1254
    [Google Scholar]
  36. Sorger G. J. 1968; Regulation of nitrogen fixation in Azotobacter vinelandii OP and in an apparently partially constitutive mutant. Journal of Bacteriology 95:1721–1726
    [Google Scholar]
  37. Strandberg G. W., Wilson P. W. 1968; Formation of the nitrogen-fixing enzyme system in Azotobacter vinelandii . Canadian Journal of Microbiology 14:25–31
    [Google Scholar]
  38. Tempest D. W., Meers J. L., Brown C. M. 1970; Influence of environment on the content and composition of microbial-free amino acid pools. Journal of General Microbiology 64:171–185
    [Google Scholar]
  39. Wilson P. W. 1958; Asymbiotic nitrogen fixation. In Encyclopedia of Plant Physiology, 8 pp. 9–47 Edited by Ruhland W. Berlin: Springer;
    [Google Scholar]
  40. Yoch D. C., Pengra R. M. 1966; Effect of amino acids on the nitrogenase system of Klebsiella pneumoniae . Journal of Bacteriology 92:618–622
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-73-2-221
Loading
/content/journal/micro/10.1099/00221287-73-2-221
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error