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Appl Environ Microbiol. 1985 May; 49(5): 1134-1141
Copyright © 1985, American Society for Microbiology. All Rights Reserved.

15N Kinetic Analysis of N2O Production by Nitrosomonas europaea: an Examination of Nitrifier Denitrification

Mark Poth and Dennis D. Focht*

2 Department of Soil and Environmental Sciences, University of California, Riverside, California 92521, 1 and Pacific Southwest Forest and Range Experiment Station, Forest Service, U.S. Department of Agriculture, Riverside, California 925072

ABSTRACT

A series of 15N isotope tracer experiments showed that Nitrosomonas europaea produces nitrous oxide only under oxygen-limiting conditions and that the labeled N from nitrite, but not nitrate, is incorporated into nitrous oxide, indicating the presence of the "denitrifying enzyme" nitrite reductase. A kinetic analysis of the m/z 44, 45, and 46 nitrous oxide produced by washed cell suspensions of N. europaea when incubated with 4 mM ammonium (99% 14N) and 0.4 mM nitrite (99% 15N) was performed. No labeled nitrite was reduced to ammonium. All labeled material added was accounted for as either nitrite or nitrous oxide. The hypothesis that nitrous oxide is produced directly from nitrification was rejected since (i) it does not allow for the large amounts of double-labeled (m/z 46) nitrous oxide observed; (ii) the observed patterns of m/z 44, 45, and 46 nitrous oxide were completely consistent with a kinetic analysis based on denitrification as the sole mechanism of nitrous oxide production but not with a kinetic analysis based on both mechanisms; (iii) the asymptotic ratio of m/z 45 to m/z 46 nitrous oxide was consistent with denitrification kinetics but inconsistent with nitrification kinetics, which predicted no limit to m/z 45 production. It is concluded that N. europaea is a denitrifier which, under conditions of oxygen stress, uses nitrite as a terminal electron acceptor and produces nitrous oxide.

FOOTNOTES

* Corresponding author.

Appl Environ Microbiol. 1985 May; 49(5): 1134-1141
Copyright © 1985, American Society for Microbiology. All Rights Reserved.



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