Skip to main content
Log in

Amperometric method for determining nitrous oxide in denitrification and in nitrogenase-catalyzed nitrous oxide reduction

  • Published:
Current Microbiology Aims and scope Submit manuscript

Abstract

A conventional Clark-type O2 probe was used to determine N2O concentrations in suspensions. At a polarizing voltage of−0.95 V versus the reference Ag/AgCl electrode, the probe is almost half as sensitive for N2O as for O2, and the detection limit is less than 1 μM N2O. The probe can also be used to determine NO for which the suitable polarizing voltage is−0.7 V. The method was successfully applied for continuously recording dissimilatory formation or utilization of N2O by intactAzospirillum brasilense Sp 7, NO production by extracts from this bacterium, and N2O reduction catalyzed by nitrogenase in intactKlebsiella pneumoniae. It is concluded that the probe is useful for measuring N2O or NO contents in bacterial suspensions when the O2 level is zero or kept constant during the assays.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

Literature Cited

  1. Bergersen FJ (1980) Measurement of nitrogen fixation by direct means. In: Bergersen FJ (ed) Methods for evaluating biological nitrogen fixation. New York: John Wiley, pp 65–110

    Google Scholar 

  2. Bothe H, Klein B, Stephan MP, Döbereiner J (1981) Transformations of inorganic nitrogen byAzospirillum spp. Arch Microbiol 130:96–100

    Google Scholar 

  3. Dalton H (1980) The cultivation of diazotrophic microorganisms. In: Bergersen FJ (ed) Methods for evaluating biological nitrogen fixation. New York: John Wiley, pp 13–64

    Google Scholar 

  4. Delwiche CC (1981) Denitrification, nitrification and atmospheric nitrous oxide. New York: John Wiley, pp 1–286

    Google Scholar 

  5. Hanus FJ, Carter KR, Evans HJ (1980) Techniques for measurement of hydrogen evolution by nodules. Methods Enzymol 69:731–739

    Google Scholar 

  6. Hardy RWF, Knight E Jr (1966) Reduction of N2O by biological N2-fixing systems. Biochem Biophys Res Commun 23:409–414

    Google Scholar 

  7. Hoch GE, Schneider KC, Burris RH (1960) Hydrogen evolution and exchange, and conversion of N2O to N2 by soybean root nodules. Biochim Biophys Acta 37:273–279

    Google Scholar 

  8. Kim C-H, Hollocher TC (1983)15N-tracer studies on the reduction of nitrite by purified dissimilatory nitrite reductase ofPseudomonas aeruginosa. J Biol Chem 258:4861–4863

    Google Scholar 

  9. Mozen MM, Burris RH (1954) The incorporation of15N-labelled nitrous oxide by nitrogen fixing agents. Biochim Biophys Acta 14:577–578

    Google Scholar 

  10. Neuer G, Bothe H (1982) The pyruvate: ferredoxin oxido-reductase in heterocysts of the cyanobacteriumAnabaena cylindrica. Biochim Biophys Acta 716:358–365

    Google Scholar 

  11. Oehme F, Schuler P (1983) Gelöst-Sauerstoff-Messung. Heidelberg: A Hütting, pp 1–146

    Google Scholar 

  12. Penteado Stephan M, Zimmer W, Bothe H (1984) Denitrification byAzospirillum brasilense Sp 7. II. Growth with nitrous oxide as respiratory electron acceptor. Arch Microbiol 138:212–216

    Google Scholar 

  13. Stephen H, stephen T (1963) Solubility of inorganic and organic compounds, vol 1. Binary systems, parts 1 and 2, Oxford: Pergamon, pp 1–1933

    Google Scholar 

  14. Steudel R, Schenk PW (1975) Stickstoffmonoxid NO. In: Bauer G (ed) Handbuch der präparativen anorganischen Chemie, vol 1. Stuttgart: Enke, pp 470–471

    Google Scholar 

  15. Wang R, Healey FR, Myers J (1971) Amperometric measurement of hydrogen evolution inChlamydomonas. Plant Physiol 48:108–110

    Google Scholar 

  16. Zimmer W, Penteado Stephan M, Bothe H (1984) Denitrification byAzospirillum brasilense Sp 7. I. Growth with nitrate as respiratory electron acceptor. Arch Microbiol 138:206–211

    Google Scholar 

  17. Albery WJ, Brooks WN, Gibson SP, Heslop MW (1979) An electroanalytical method for the determination of N2O. Electrochim Acta 24:107–108

    Google Scholar 

  18. Alefounder PR, Ferguson SJ (1982) Electron transport linked nitrous oxide synthesis and reduction byParacoccus denitrificans monitored with an electrode. Biochem Biophys Res Commun 104:1149–1155

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zimmer, W., Danneberg, G. & Bothe, H. Amperometric method for determining nitrous oxide in denitrification and in nitrogenase-catalyzed nitrous oxide reduction. Current Microbiology 12, 341–345 (1985). https://doi.org/10.1007/BF01567894

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01567894

Keywords

Navigation