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Journal of Bacteriology logoLink to Journal of Bacteriology
. 1997 Feb;179(4):1362–1367. doi: 10.1128/jb.179.4.1362-1367.1997

Nitrogenase activity and regeneration of the cellular ATP pool in Azotobacter vinelandii adapted to different oxygen concentrations.

K Linkerhägner 1, J Oelze 1
PMCID: PMC178837  PMID: 9023223

Abstract

The in vivo activity of nitrogenase under aerobiosis was studied with diazotrophic chemostat cultures of Azotobacter vinelandii grown under glucose- or phosphate-limited conditions at different dilution rates (Ds, representing the growth rate mu) and different dissolved oxygen concentrations. Under steady-state conditions, the concentration as well as the cellular level of ATP increased in glucose-limited cultures when D was increased. Irrespective of the type of growth limitation or the dissolved oxygen concentration, the steady-state concentrations of ATP and of dinitrogen fixed by nitrogenase increased in direct proportion to each other. Specific rates of dinitrogen fixation as well as of the regeneration of the cellular ATP pool were compared with specific rates of cellular respiration. With glucose-limited cultures, the rate of regeneration of the ATP pool and the rate of respiration varied in direct proportion to each other. This relationship, however, was dependent on the dissolved oxygen concentration. As compared to the phosphate-sufficient control, phosphate-limited cultures exhibited the same nitrogenase activity but significantly increased respiratory activities. Rates of ATP regeneration and of cellular respiration of phosphate-limited cultures did not fit into the relationship characteristic of glucose-limited cultures. However, a linear relationship between the rates of dinitrogen fixation and ATP regeneration was identified irrespective of the type of growth limitation and the dissolved oxygen concentration. The results suggest that the ATP supply rather than cellular oxygen consumption is of primary importance in keeping nitrogenase activity in aerobic cultures of A. vinelandii.

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Selected References

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  1. Dalton H., Postgate J. R. Effect of oxygen on growth of Azotobacter chroococcum in batch and continuous cultures. J Gen Microbiol. 1968 Dec;54(3):463–473. doi: 10.1099/00221287-54-3-463. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Dingler C., Kuhla J., Wassink H., Oelze J. Levels and activities of nitrogenase proteins in Azotobacter vinelandii grown at different dissolved oxygen concentrations. J Bacteriol. 1988 May;170(5):2148–2152. doi: 10.1128/jb.170.5.2148-2152.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Drozd J., Postgate J. R. Effects of oxygen on acetylene reduction, cytochrome content and respiratory activity of Azotobacter chroococcum. J Gen Microbiol. 1970 Sep;63(1):63–73. doi: 10.1099/00221287-63-1-63. [DOI] [PubMed] [Google Scholar]
  5. Haaker H., Veeger C. Involvement of the cytoplasmic membrane in nitrogen fixation by Azotobacter vinelandii. Eur J Biochem. 1977 Jul 1;77(1):1–10. doi: 10.1111/j.1432-1033.1977.tb11634.x. [DOI] [PubMed] [Google Scholar]
  6. Kelly M. J., Poole R. K., Yates M. G., Kennedy C. Cloning and mutagenesis of genes encoding the cytochrome bd terminal oxidase complex in Azotobacter vinelandii: mutants deficient in the cytochrome d complex are unable to fix nitrogen in air. J Bacteriol. 1990 Oct;172(10):6010–6019. doi: 10.1128/jb.172.10.6010-6019.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kuhla J., Oelze J. Dependence of nitrogenase switch-off upon oxygen stress on the nitrogenase activity in Azotobacter vinelandii. J Bacteriol. 1988 Nov;170(11):5325–5329. doi: 10.1128/jb.170.11.5325-5329.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  9. Lees H., Postgate J. R. The behaviour of Azotobacter chroococcum in oxygen- and phosphate-limited chemostat culture. J Gen Microbiol. 1973 Mar;75(1):161–166. doi: 10.1099/00221287-75-1-161. [DOI] [PubMed] [Google Scholar]
  10. Linkerhägner K., Oelze J. Cellular ATP levels and nitrogenase switchoff upon oxygen stress in chemostat cultures of Azotobacter vinelandii. J Bacteriol. 1995 Sep;177(18):5289–5293. doi: 10.1128/jb.177.18.5289-5293.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Linkerhägner K., Oelze J. Hydrogenase does not confer significant benefits to Azotobacter vinelandii growing diazotrophically under conditions of glucose limitation. J Bacteriol. 1995 Oct;177(20):6018–6020. doi: 10.1128/jb.177.20.6018-6020.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lundin A., Thore A. Comparison of methods for extraction of bacterial adenine nucleotides determined by firefly assay. Appl Microbiol. 1975 Nov;30(5):713–721. doi: 10.1128/am.30.5.713-721.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Pirt S. J. Maintenance energy: a general model for energy-limited and energy-sufficient growth. Arch Microbiol. 1982 Dec 3;133(4):300–302. doi: 10.1007/BF00521294. [DOI] [PubMed] [Google Scholar]
  14. Post E., Kleiner D., Oelze J. Whole cell respiration and nitrogenase activities in Azotobacter vinelandii growing in oxygen controlled continuous culture. Arch Microbiol. 1983 Jan;134(1):68–72. doi: 10.1007/BF00429410. [DOI] [PubMed] [Google Scholar]
  15. Russell J. B., Cook G. M. Energetics of bacterial growth: balance of anabolic and catabolic reactions. Microbiol Rev. 1995 Mar;59(1):48–62. doi: 10.1128/mr.59.1.48-62.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Scherings G., Haaker H., Wassink H., Veeger C. On the formation of an oxygen-tolerant three-component nitrogenase complex from Azotobacter vinelandii. Eur J Biochem. 1983 Oct 3;135(3):591–599. doi: 10.1111/j.1432-1033.1983.tb07693.x. [DOI] [PubMed] [Google Scholar]
  17. Tempest D. W., Neijssel O. M. The status of YATP and maintenance energy as biologically interpretable phenomena. Annu Rev Microbiol. 1984;38:459–486. doi: 10.1146/annurev.mi.38.100184.002331. [DOI] [PubMed] [Google Scholar]
  18. Thorneley R. N., Ashby G. A. Oxidation of nitrogenase iron protein by dioxygen without inactivation could contribute to high respiration rates of Azotobacter species and facilitate nitrogen fixation in other aerobic environments. Biochem J. 1989 Jul 1;261(1):181–187. doi: 10.1042/bj2610181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Tsai J. C., Aladegbami S. L., Vela G. R. Phosphate-limited culture of Azotobacter vinelandii. J Bacteriol. 1979 Aug;139(2):639–645. doi: 10.1128/jb.139.2.639-645.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Wang Z. C., Burns A., Watt G. D. Complex formation and O2 sensitivity of Azotobacter vinelandii nitrogenase and its component proteins. Biochemistry. 1985 Jan 1;24(1):214–221. doi: 10.1021/bi00322a031. [DOI] [PubMed] [Google Scholar]
  21. Yates M. G. Control of respiration and nitrogen fixation by oxygen and adenine nucleotides in N2-grown Azotobacter chroococcum. J Gen Microbiol. 1970 Mar;60(3):393–401. doi: 10.1099/00221287-60-3-393. [DOI] [PubMed] [Google Scholar]

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