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The pathway of nitrogen and reductive enzymes of denitrification

  • Denitrification: Recent Advances and Future Directions
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Abstract

Some recent studies on the pathway of nitrogen and the reductases of denitrification are reviewed. The available evidence suggests that while the intermediates of denitrification can remain enzyme-bound (presumably to nitrite reductase) prior to formation of N2O, NO and nitroxyl (HNO) can be released in part by certain bacteria. Release of NO is recognized by a nitrite/NO−15N exchange reaction and isotopic scrambling in product N2O; release of nitroxyl by Pseudomonas stutzeri is recognized by isotopic scrambling of nitrite and NO in product N2O in absence of exchange and affords evidence that the first N−N bond forms in denitrification at the N1+ redox level. The recent purification and partial characterization of nitrous oxide reductase are described. The ability of the dissimilatory nitrite reductase to activate nitrite for nitrosyl transfer affords a new chemical probe into the mechanism of action of this central enzyme. It would appear that reduction of nitrite is subject to electrophilic catalysis. 18O studies show that dissociation of nitrite from nitrite reductase can be slow relative to competing reduction or nitrosyl transfer.

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References

  • Averill, B. A. and Tiedje, J. M. 1982. The chemical mechanism of microbial denitrification. —FEBS Lett. 138: 8–12.

    Google Scholar 

  • Balderston, W. L., Sherr, B. and Payne, W. M. 1976. Blockage by acetylene of nitrous oxide reduction in Pseudomonas perfectomarinus. — Appl. Environ. Microbiol. 31: 504–508.

    Google Scholar 

  • Betlach, M. R. and Tiedje, J. M. 1981. Kinetic explanation for accumulation of nitrite, nitric oxide and nitrous oxide during bacterial denitrification. — Appl. Environ. Microbiol. 42: 1074–1084.

    Google Scholar 

  • Bonner, F. T. and Akhtar, M. J. 1981. Formation of nitrosyltricyanonickelate (NiNO(CN) 2-3 ) in a direct NO- displacement reaction. — Inorg. Chem. 20: 3155–3160.

    Google Scholar 

  • Bonner, F. T., Dzelzkains, L. S. and Bonucci, J. A. 1978. Properties of nitroxyl as intermediate in the nitric oxide-hydroxylamine reaction and in trioxydinitrite decomposition. — Inorg. Chem. 17: 2487–2494.

    Google Scholar 

  • Bothner-By, A. and Friedman, L. 1952. The reaction of nitrous acid with hydroxylamine. — J. Chem. Phys. 20: 459–462.

    Google Scholar 

  • Castignetti, D. and Gunner, H. B. 1981. Nitrite and nitrate synthesis from pyruvic oxime by an Alcaligenes sp. — Curr. Microbiol. 5: 379–384.

    Google Scholar 

  • Castignetti, D. and Hollocher, T. C. 1981. Vigorous denitrification by a heterotrophic nitrifier of the genus Alcaligenes. — Curr. Microbiol. 6: 229–231.

    Google Scholar 

  • Cooper, J. N., Chilton, J. E. and Powell, R. E. 1970. Reaction of nitric oxide with alkaline hydroxylamine. — Inorg. Chem. 9: 2303–2304.

    Google Scholar 

  • Fewson, C. A. and Nicholas, D. J. D. 1960. Utilization of nitric oxide by microorganisms and higher plants. — Nature (London) 188: 794–796.

    Google Scholar 

  • Firestone, M. K., Firestone, R. B. and Tiedje, J. M. 1979. Nitric oxide as an intermediate in denitrification: Evidence from nitrogen-13 isotope exchange. — Biochem. Biophys. Res. Commun. 91: 10–16.

    Google Scholar 

  • Garber, E. and Hollocher, T. C. 1980. 15N-isotope studies with denitrifying bacteria. — Fed. Proc. 39: 1773 (Abstract).

    Google Scholar 

  • Garber, E. A. E. and Hollocher, T. C., 1981. 15N Tracer studies on the role of NO in denitrification. — J. Biol. Chem. 256: 5459–5465.

    Google Scholar 

  • Garber, E. A. E. and Hollocher, T. C. 1982a. Positional isotopic equivalence of nitrogen in N2O produced by the denitrifying bacterium Pseudomonas stutzeri. Indirect evidence for a nitroxyl pathway. — J. Biol. Chem. 257: 4705–4708.

    Google Scholar 

  • Garber, E. A. E. and Hollocher, T. C. 1982b. 15N, 18O-tracer studies on the activation of nitrite by denitrifying bacteria. Nitrite/water-oxygen exchange and nitrosation reactions as indicators of electrophilic catalysis. — J. Biol. Chem. 257: 8091–8097.

    Google Scholar 

  • Hollocher, T. C., Garber, E., Cooper, A. J. L. and Reiman, R. E. 1980. 13N, 15N Isotope and kinetic evidence against hyponitrite as an intermediate in denitrification. — J. Biol. Chem. 255: 5027–5030.

    Google Scholar 

  • Iwasaki, H., Saigo, T. and Matsubara, T. 1980. Copper as a controlling factor of anaerobic growth under N2O and biosynthesis of N2O reductase in denitrifying bacteria. — Plant Cell Physiol. 21: 1573–1584.

    Google Scholar 

  • Kristjansson, J. K. 1980. N-oxide respiration and the enzymology of nitrous oxide reduction in denitrifiers. — Ph.D. Dissertation, Brandeis University, Waltham MA, USA.

  • Kristjansson, J. K. and Hollocher, T. C. 1980. First practical assay for soluble nitrous oxide reductase of denitrifying bacteria and a partial kinetic characterization. — J. Biol. Chem. 255: 704–707.

    Google Scholar 

  • Kristjansson, J. K. and Hollocher, T. C. 1981. Partial purification and characterization of nitrous oxide reductase from Paracoccus denitrificans. — Curr. Microbiol. 6: 247–251.

    Google Scholar 

  • Matsubara, T., Frunzke, K. and Zumft, W. G. 1982. Modulation by copper of the products of nitrite respiration in Pseudomonas perfectomarinus. — J. Bacteriol. 149: 816–823.

    Google Scholar 

  • Matsubara, T. and Iwasaki, H. 1972. Nitric oxide-reducing activity of Alcaligenes faecalis cytochrome cd. — J. Biochem. (Tokyo) 72: 57–64.

    Google Scholar 

  • Miyata, M. 1971. Studies on denitrification. XIV. The electron donating system in the reduction of nitric oxide and nitrate. — J. Biochem. (Tokyo) 70: 205–213.

    Google Scholar 

  • Payne, W. J., Riley, P. S. and Cox, C. D. 1971. Separate nitrite, nitric oxide, and nitrous oxide reducing fractions from Pseudomonas perfectomarinus. — J. Bacteriol. 106: 356–361.

    Google Scholar 

  • St. John, R. T. and Hollocher, T. C. 1977. Nitrogen 15 tracer studies on the pathway of denitrification in Pseudomonas aeruginosa. — J. Biol. Chem. 252: 212–218.

    Google Scholar 

  • Taylor, R. C., Brown, R. A., Young, W. S. and Headington, C. F. 1948. The mass spectrometer in organic chemical analysis. — Anal. Chem. 20: 396–401.

    Google Scholar 

  • Wharton, D. C. and Weintraub, S. T. 1980. Identification of nitric oxide and nitrous oxide as products of nitrite reduction by Pseudomonas cytochrome oxidase (nitrite reductase). — Biochem. Biophys. Res. Commun. 97: 236–242.

    Google Scholar 

  • Yoshinari, T. and Knowles, R. 1976. Acetylene inhibition of nitrous oxide reduction by denitrifying bacteria. — Biochem. Biophys. Res. Commun. 69: 705–710.

    Google Scholar 

  • Zumft, W. G. and Vega, J. M. 1979. Reduction of nitrite to nitrous oxide by a cytoplasmic membrane fraction from the marine denitrifier Pseudomonas perfectomarinus. — Biochim. Biophys. Acta 548: 484–499.

    Google Scholar 

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Hollocher, T.C. The pathway of nitrogen and reductive enzymes of denitrification. Antonie van Leeuwenhoek 48, 531–544 (1983). https://doi.org/10.1007/BF00399539

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