1887

Microbiology Society logo

Volume 139, Issue 6

Oxygen inhibition of nitrogenase activity in Free

Abstract

A purpose-built oxystat has been used to study reversible inhibition of nitrogenase by O in the facultative anaerobe CH-reducing activity in samples from either an anaerobic glucose-limited or an O-limited diazotrophic chemostat culture was completely inhibited by exposure to a dissolved O concentration (DOC) of 1·5 μm or above. Subsequently, under anaerobic conditions, CH-reducing activity returned in the absence of protein synthesis. The amount of activity returning never reached 100% of the initial anaerobic activity before O treatment. The degree of reversibility was inversely proportional to the log of DOC during exposure and was decreased by increasing the time of exposure to O (about 60% reversibility occurred after a 20 min exposure to 6 μm-O). The failure to obtain complete recovery of activity was apparently not due to inactivation of the very O-sensitive pyruvate-flavodoxin oxidoreductase ( product) which provides electrons for nitrogenase activity Samples from the O-limited culture behaved similarly to those limited by glucose. Thus, ‘training’ of the organism to use O during growth does not influence the tolerance of nitrogenase to O. Since the behaviour towards O reported here for differs from that known to occur in , the mechanism of protection of nitrogenase from O damage may differ in these organisms.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
© Society for General Microbiology 1993
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-139-6-1307
1993-06-01
2024-04-18
Loading full text...

Full text loading...

/deliver/fulltext/micro/139/6/mic-139-6-1307.html?itemId=/content/journal/micro/10.1099/00221287-139-6-1307&mimeType=html&fmt=ahah

References

  1. Bergersen F. J., Turner G. L. 1979; Systems utilising oxygenated leghaemoglobin and myoglobin as sources of free dissolved O2at low concentrations for experiments with bacteria.. Analytical Biochemistry 96165–174
     [Google Scholar]
  2. Cannon F. C. 1980; Genetic studies with diazotrophs.. in Methods for Evaluating Biological Nitrogen Fixation pp. 367–413 Bergersen F. J. Edited by Chichester: John Wiley;
     [Google Scholar]
  3. Cannon M., Hill S., Kavanagh E., Cannon F. 1985; A molecular study ofnifexpression inKlebsiella pneumoniaeat the level of transcription, translation and nitrogenase activity.. Molecular and General Genetics 198:198–206
     [Google Scholar]
  4. Deistung J., Cannon F. C., Cannon M. C., Hill S., Thorneley R. N. F. 1985; Electron transfer to nitrogenase inKlebsiella pneumoniae:nifFgene cloned and the gene product, a flavodoxin purified.. Biochemical Journal 231:743–753
     [Google Scholar]
  5. Dingler Ch., Oelze J. 1985; Reversible and irreversible inactivation of cellular nitrogenase upon oxygen stress inAzotobacter vinelandiigrowing in oxygen controlled continuous culture.. Archives of Microbiology 141:80–84
     [Google Scholar]
  6. Drozd J., Postgate J. R. 1970; Effects of oxygen on acetylene reduction, cytochrome content and respiratory activity ofAzotobacter chroococcum. . Journal of General Microbiology 63:63–73
     [Google Scholar]
  7. Drummond M. 1986; Structure predictions and surface change of nitrogenase flavodoxins fromKlebsiella pneumoniaeandAzotobacter vinelandii. . European Journal of Biochemistry 159:549–553
     [Google Scholar]
  8. Goldberg I., Nadler V., Hochman A. 1987; Mechanism of nitrogenase ʻswitch off̕ by oxygen.. Journal of Bacteriology 169:874–879
     [Google Scholar]
  9. Haaker H., Veeger C. 1977; Involvement of the cytoplasmic membrane in nitrogen fixation byAzotobacter vinelandii. . European Journal of Biochemistry 77:1–10
     [Google Scholar]
  10. Hill S. 1976a; Influence of atmospheric oxygen concentration on acetylene reduction and efficiency of nitrogen fixation in intactKlebsiella pneumoniae. . Journal of General Microbiology 93:335–345
     [Google Scholar]
  11. Hill S. 1976b; The apparent ATP requirement for nitrogen fixation in growingKlebsiella pneumoniae. . Journal of General Microbiology 95:297–312
     [Google Scholar]
  12. Hill S., Kavanagh E. 1980; Roles ofnifFandnifJgene products in the electron transport to nitrogenase inKlebsiella pneumoniae. . Journal of Bacteriology 141:470–475
     [Google Scholar]
  13. Hill S., Turner G. L., Bergersen F. J. 1984; Synthesis and activity of nitrogenase inKlebsiella pneumoniaeexposed to low concentrations of oxygen.. Journal of General Microbiology 130:1061–1067
     [Google Scholar]
  14. Hill S., Viollet S., Smith A., Anthony C. 1990; Roles for entericd-type cytochrome oxidase inN2 fixation and micro- aerobiosis.. Journal of Bacteriology 172:2071–2078
     [Google Scholar]
  15. Hochman A., Goldberg I., Nadler V., Hartmann A. 1987; The mechanism of nitrogenase reversible ʻswitch off̕ by oxygen.. In Inorganic Nitrogen Metabolism pp. 173–174 Ullrich W. R., Aparicio P. J., Syreet P. J., Castillo F. Edited by Berlin: Springer Verlag;
     [Google Scholar]
  16. Johnson M. J., Borkowski J., Engelblom C. 1964; Steam sterilisable probes for dissolved oxygen measurements.. Biotechnology and Bioengineering 6:457–468
     [Google Scholar]
  17. Kavanagh E., Hill S. 1990; The automatic maintenance of low dissolved oxygen using a photobacterial oxygen sensor for the study of microaerobiosis.. Journal of Applied Bacteriology 69:539–549
     [Google Scholar]
  18. Kelly M. J. S., Poole R. K., Yates M. G., Kennedy C. 1990; Cloning and mutagenesis of genes encoding the cytochromebdterminal oxidase complex inAzotobacter vinelandii.Mutants deficient in the cytochromedcomplex are unable to fix nitrogen in air.. Journal of Bacteriology 172:6010–6019
     [Google Scholar]
  19. Nieva-Gomez D., Roberts G. P., Klevickis S., Brill W. J. 1980; Electron transport to nitrogenase inKlebsiella pneumoniae. . Proceedings of the National Academy of Sciences of the United States of America 77:2555–2558
     [Google Scholar]
  20. Robson R. L., Postgate J. R. 1980; Oxygen and hydrogen in biological nitrogen fixation.. Annual Review of Microbiology 34:183–207
     [Google Scholar]
  21. Scherings G., Haaker H., Veeger C. 1977; Regulation of nitrogen fixation by Fe-S protein II, inAzotobacter vinelandii. . European Journal of Biochemistry 77:621–630
     [Google Scholar]
  22. Shah V. K., Stacey G., Brill W. J. 1983; Electron transport to nitrogenase. Purification and characterisation of pyruvate- flavodoxin oxidoreductase, thenifJproduct.. Journal of Biological Chemistry 258:12064–12068
     [Google Scholar]
  23. Smith A., Hill S., Anthony C. 1990; The purification, characterization and role of the rf-type cytochrome oxidase ofKlebsiella pneumoniaeduring nitrogen fixation.. Journal of General Microbiology 136:171–180
     [Google Scholar]
  24. Tubb R. S., Postgate J. R. 1973; Control of nitrogenase synthesis inKlebsiella pneumoniae. . Journal of General Microbiology 79:103–117
     [Google Scholar]
  25. Thorneley R. N. F., Ashby G. A. 1989; Oxidation of nitrogenase iron protein by dioxygen without inactivation could contribute to high respiration rates ofAzotobacterspecies and facilitate nitrogen fixation in other aerobic environments.. Biochemical Journal 261:181–187
     [Google Scholar]
  26. Thorneley R.N. F., Deistung J. 1988; Electron-transfer studies involving flavodoxin and a natural redox partner, the iron protein of nitrogenase. Conformational constraints on protein-protein interactions and the kinetics of electron transfer within the protein complex.. Biochemical Journal 253:587–595
     [Google Scholar]
  27. Thorneley R. N. F., Abell C., Ashby G. A., Drummond M. H., Eady R. R., Huff S., Macdonald C. J., Shneier A. 1992; Posttranslational modifications ofKlebsiella pneumoniaeflavodoxin by covalent attachment of coenzyme A, shown by31P NMR and electrospray mass spectrometry, prevents electron transfer from thenifJprotein to nitrogenase. A possible new regulatory mechanism for biological nitrogen fixation.. Biochemistry 31:1216–1224
     [Google Scholar]
  28. Wahl R. C., Orme-Johnson W. H. 1987; Clostridial pyruvate oxidoreductase and the pyruvate oxidising enzyme specific to nitrogen fixation inKlebsiella pneumoniaeare similar enzymes.. Journal of Biological Chemistry 282:10489–10496
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-139-6-1307
Loading
Oxygen inhibition of nitrogenase activity in Klebsiella pneumoniae
Microbiology 139, 1307 (1993); https://doi.org/10.1099/00221287-139-6-1307
/content/journal/micro/10.1099/00221287-139-6-1307
/content/journal/micro/10.1099/00221287-139-6-1307
Loading

Data & Media loading...

Most cited Most Cited RSS feed

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