Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1971 Mar;105(3):913–917. doi: 10.1128/jb.105.3.913-917.1971

Nitrogenase Activity in Cell-Free Extracts of the Blue-Green Alga, Anabaena cylindrica

R V Smith 1, M C W Evans 1
PMCID: PMC248518  PMID: 4994040

Abstract

Cell-free extracts with high nitrogenase activity were prepared by sonic oscillation and French press treatment from the blue-gree alga Anabaena cylindrica. Extracts were prepared from cells grown on a 95% N2–5% CO2 gas mixture followed by a period of nitrogen starvation under an atmosphere of 95% argon–5% CO2. No increase in the specific activity of extracts was achieved by breaking heterocysts. Activity (assayed by acetylene reduction) was found to be dependent on adenosine triphosphate (ATP), an ATP-generating system, and a low-potential reductant. Na2S2O2 employed as reductant supports higher rates of nitrogenase activity than reduced ferredoxin. The activity is associated with a small-particle fraction that can be sedimented by ultracentrifugation. In contrast to the particulate nitrogenase of Azotobacter, which is stable in air, the A. cylindrica nitrogenase is an oxygen sensitive as nitrogenase prepared from anaerobic bacteria.

Full text

PDF
913

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Allen M. B., Arnon D. I. Studies on Nitrogen-Fixing Blue-Green Algae. I. Growth and Nitrogen Fixation by Anabaena Cylindrica Lemm. Plant Physiol. 1955 Jul;30(4):366–372. doi: 10.1104/pp.30.4.366. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Arnon D. I. COPPER ENZYMES IN ISOLATED CHLOROPLASTS. POLYPHENOLOXIDASE IN BETA VULGARIS. Plant Physiol. 1949 Jan;24(1):1–15. doi: 10.1104/pp.24.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Biggins D. R., Postgate J. R. Nitrogen fixation by cultures and cell-free extracts of Mycobacterium flavum 301. J Gen Microbiol. 1969 May;56(2):181–193. doi: 10.1099/00221287-56-2-181. [DOI] [PubMed] [Google Scholar]
  4. Bulen W. A., Burns R. C., LeComte J. R. Nitrogen fixation: cell-free system with extracts of Azotobacter. Biochem Biophys Res Commun. 1964 Oct 14;17(3):265–271. doi: 10.1016/0006-291x(64)90395-x. [DOI] [PubMed] [Google Scholar]
  5. Bulen W. A., LeComte J. R. The nitrogenase system from Azotobacter: two-enzyme requirement for N2 reduction, ATP-dependent H2 evolution, and ATP hydrolysis. Proc Natl Acad Sci U S A. 1966 Sep;56(3):979–986. doi: 10.1073/pnas.56.3.979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Burris R. H. Progress in the biochemistry of nitrogen fixation. Proc R Soc Lond B Biol Sci. 1969 Apr 1;172(1029):339–354. doi: 10.1098/rspb.1969.0025. [DOI] [PubMed] [Google Scholar]
  7. CARNAHAN J. E., MORTENSON L. E., MOWER H. F., CASTLE J. E. Nitrogen fixation in cell-free extracts of Clostridium pasteurianum. Biochim Biophys Acta. 1960 Nov 18;44:520–535. doi: 10.1016/0006-3002(60)91606-1. [DOI] [PubMed] [Google Scholar]
  8. COX R. M., FAY P., FOGG G. E. NITROGEN FIXATION AND PHOTOSYNTHESIS IN A SUBCELLULAR FRACTION OF THE BLUE-GREEN ALGA ANABAENA CYLINDRICA. Biochim Biophys Acta. 1964 Jul 29;88:208–210. doi: 10.1016/0926-6577(64)90168-8. [DOI] [PubMed] [Google Scholar]
  9. Dalton H., Postgate J. R. Growth and physiology of Azotobacter chroococcum in continuous culture. J Gen Microbiol. 1969 Jun;56(3):307–319. doi: 10.1099/00221287-56-3-307. [DOI] [PubMed] [Google Scholar]
  10. Dilworth M. J. Acetylene reduction by nitrogen-fixing preparations from Clostridium pasteurianum. Biochim Biophys Acta. 1966 Oct 31;127(2):285–294. doi: 10.1016/0304-4165(66)90383-7. [DOI] [PubMed] [Google Scholar]
  11. Evans M. C., Hall D. O., Bothe H., Whatley F. R. The stoicheiometry of electron transfer by bacterial and plant ferredoxins. Biochem J. 1968 Dec;110(3):485–489. doi: 10.1042/bj1100485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fay P., Cox R. M. Oxygen inhibition of nitrogen fixation in cell-free preparations of blue-green algae. Biochim Biophys Acta. 1967;143(3):562–569. doi: 10.1016/0005-2728(67)90061-8. [DOI] [PubMed] [Google Scholar]
  13. Fay P., Stewart W. D., Walsby A. E., Fogg G. E. Is the heterocyst the site of nitrogen fixation in blue-green algae? Nature. 1968 Nov 23;220(5169):810–812. doi: 10.1038/220810b0. [DOI] [PubMed] [Google Scholar]
  14. Fay P., Walsby A. E. Metabolic activities of isolated heterocysts of the blue-green alga Anabaena cylindrica. Nature. 1966 Jan 1;209(5018):94–95. doi: 10.1038/209094a0. [DOI] [PubMed] [Google Scholar]
  15. Hardy R. W., Holsten R. D., Jackson E. K., Burns R. C. The acetylene-ethylene assay for n(2) fixation: laboratory and field evaluation. Plant Physiol. 1968 Aug;43(8):1185–1207. doi: 10.1104/pp.43.8.1185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kelly M. Comparisons and cross reactions of nitrogenase from Klebsiella pneumoniae, Azotobacter chroococcum and Bacillus polymyxa. Biochim Biophys Acta. 1969;191(3):527–540. doi: 10.1016/0005-2744(69)90346-5. [DOI] [PubMed] [Google Scholar]
  17. RABINOWITZ J. C., PRICER W. E., Jr Formyltetrahydrofolate synthetase. I. Isolation and crystallization of the enzyme. J Biol Chem. 1962 Sep;237:2898–2902. [PubMed] [Google Scholar]
  18. Schneider K. C., Bradbeer C., Singh R. N., Wang L. C., Wilson P. W., Burris R. H. NITROGEN FIXATION BY CELL-FREE PREPARATIONS FROM MICROORGANISMS. Proc Natl Acad Sci U S A. 1960 May;46(5):726–733. doi: 10.1073/pnas.46.5.726. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Smith R. V., Evans M. C. Soluble nitrogenase from vegetative cells of the blue-green alga Anabaena cylindrica. Nature. 1970 Mar 28;225(5239):1253–1254. doi: 10.1038/2251253a0. [DOI] [PubMed] [Google Scholar]
  20. Stewart W. D. Biological and ecological aspects of nitrogen fixation by free-living micro-organisms. Proc R Soc Lond B Biol Sci. 1969 Apr 1;172(1029):367–388. doi: 10.1098/rspb.1969.0027. [DOI] [PubMed] [Google Scholar]
  21. Stewart W. D., Haystead A., Pearson H. W. Nitrogenase activity in heterocysts of blue-green algae. Nature. 1969 Oct 18;224(5216):226–228. doi: 10.1038/224226a0. [DOI] [PubMed] [Google Scholar]
  22. Wolk C. P. Movement of carbon from vegetative cells to heterocysts in Anabaena cylindrica. J Bacteriol. 1968 Dec;96(6):2138–2143. doi: 10.1128/jb.96.6.2138-2143.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wyatt J. T., Silvey J. K. Nitrogen fixation by gloeocapsa. Science. 1969 Aug 29;165(3896):908–909. doi: 10.1126/science.165.3896.908. [DOI] [PubMed] [Google Scholar]
  24. Yoch D. C., Arnon D. I. The nitrogen fixation system of photosynthetic bacteria. II. Chromatium nitrogenase activity linked to photochemically generated assimilatory power. Biochim Biophys Acta. 1970 Mar 3;197(2):180–184. doi: 10.1016/0005-2728(70)90029-0. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES