Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1982 Oct;152(1):72–80. doi: 10.1128/jb.152.1.72-80.1982

Nitrogen fixation system of tungsten-resistant mutants of Azotobacter vinelandii.

G D Riddle, J G Simonson, B J Hales, H D Braymer
PMCID: PMC221376  PMID: 6956567

Abstract

Mutants of Azotobacter vinelandii ATCC 12837 were isolated which could fix N2 in the presence of high tungsten concentrations. The most studied of these mutants (WD2) grew well in N-free modified Burk broth containing 10 mM W, whereas the wild type would not grow in this medium. WD2 would also grow in Burk N-free broth at about the same rate as the wild type. WD2 in broth containing W exhibited 22% of the whole cell acetylene reduction activity of the wild type in broth containing Mo and showed a lowered affinity for acetylene. Two-dimensional gel electrophoresis experiments showed that N2-fixing cells of WD2 from broth containing W or Mo did not produce significant amounts of component I of native nitrogenase protein. Electron spin resonance spectra of whole cells and cell-free extracts of WD2 from broth containing W lacked any trace of the g = 3.6 resonance associated with FeMoCo.

Full text

PDF
72

Images in this article

Selected References

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

  1. Benemann J. R., Smith G. M., Kostel P. J., McKenna C. E. Tungsten incorporation into Azotobacter vinelandii nitrogenase. FEBS Lett. 1973 Feb 1;29(3):219–221. doi: 10.1016/0014-5793(73)80023-7. [DOI] [PubMed] [Google Scholar]
  2. Bishop P. E., Jarlenski D. M., Hetherington D. R. Evidence for an alternative nitrogen fixation system in Azotobacter vinelandii. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7342–7346. doi: 10.1073/pnas.77.12.7342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bulen W. A. EFFECT OF TUNGSTATE ON THE UPTAKE AND FUNCTION OF MOLYBDATE IN AZOTOBACTER AGILIS. J Bacteriol. 1961 Jul;82(1):130–134. doi: 10.1128/jb.82.1.130-134.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cardenas J., Mortenson L. E. Determination of molybdenum and tungsten in biological materials. Anal Biochem. 1974 Aug;60(2):372–381. doi: 10.1016/0003-2697(74)90244-9. [DOI] [PubMed] [Google Scholar]
  5. Davis L. C., Shah V. K., Brill W. J., Orme-Johnson W. H. Nitrogenase. II. Changes in the EPR signal of component I (iron-molybdenum protein) of Azotobacter vinelandii nitrogenase during repression and derepression. Biochim Biophys Acta. 1972 Feb 28;256(2):512–523. doi: 10.1016/0005-2728(72)90079-5. [DOI] [PubMed] [Google Scholar]
  6. KEELER R. F., VARNER J. E. Tungstate as an antagonist of molybdate in Azotobacter vinelandii. Arch Biochem Biophys. 1957 Aug;70(2):585–590. doi: 10.1016/0003-9861(57)90146-7. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Nagatani H. H., Brill W. J. Nitrogenase V. The effect of Mo, W and V on the synthesis of nitrogenase components in Azotobacter vinelandii. Biochim Biophys Acta. 1974 Aug 7;362(1):160–166. doi: 10.1016/0304-4165(74)90037-3. [DOI] [PubMed] [Google Scholar]
  9. Nagatani H. H., Shah V. K., Brill W. J. Activation of inactive nitrogenase by acid-treated component I. J Bacteriol. 1974 Nov;120(2):697–701. doi: 10.1128/jb.120.2.697-701.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  11. Pienkos P. T., Klevickis S., Brill W. J. In vitro activation of inactive nitrogenase component I with molybdate. J Bacteriol. 1981 Jan;145(1):248–256. doi: 10.1128/jb.145.1.248-256.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. ROBRISH S. A., MARR A. G. Location of enzymes in Azotobacteragilis. J Bacteriol. 1962 Jan;83:158–168. doi: 10.1128/jb.83.1.158-168.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Rawlings J., Shah V. K., Chisnell J. R., Brill W. J., Zimmermann R., Münck E., Orme-Johnson W. H. Novel metal cluster in the iron-molybdenum cofactor of nitrogenase. Spectroscopic evidence. J Biol Chem. 1978 Feb 25;253(4):1001–1004. [PubMed] [Google Scholar]
  14. Singh H. N., Vaishampayan A., Singh R. K. Evidence for the involvement of a genetic determinant controlling functional specificity of group VI B elements in the metabolism of N2 and NO-3 in the blue-green alga Nostoc muscorum. Biochem Biophys Res Commun. 1978 Mar 15;81(1):67–74. doi: 10.1016/0006-291x(78)91631-5. [DOI] [PubMed] [Google Scholar]
  15. TAKAHASHI H., NASON A. Tungstate as competitive inhibitor of molybdate in nitrate assimilation and in N2 fixation by Azotobacter. Biochim Biophys Acta. 1957 Feb;23(2):433–435. doi: 10.1016/0006-3002(57)90351-7. [DOI] [PubMed] [Google Scholar]

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

RESOURCES