Hydroxylamine oxidation and subsequent nitrous oxide production by the heterotrophic ammonia oxidizer Alcaligenes faecalis.
Nitrous oxide (N2O), a greenhouse gas, is emitted during autotrophic and heterotrophic ammonia oxidation. This emission may result from either coupling to aerobic denitrification, or it may be formed in the oxidation of hydroxylamine (NH2OH) to nitrite (NO2(-). Therefore, the N2O production during NH2OH oxidation was studied with Alcaligenes faecalis strain TUD. Continuous cultures of A. faecalis showed increased N2O production when supplemented with increasing NH2OH concentrations. 15N-labeling experiments showed that this N2O production was not due to aerobic denitrification of NO2(-). Addition of 15N-labeled NH2OH indicated that N2O was a direct by-product of NH2OH oxidation, which was subsequently reduced to N2. These observations are sustained by the fact that NO2(-) production was low (0.23 mM maximum) and did not increase significantly with increasing NH2OH concentration in the feed. The NH2OH-oxidizing capacity increased with increasing NH2OH concentrations. The apparent Vmax and K(m) were 31 nmol min-1 mg dry weight-1 and 1.5 mM respectively. The culture did not increase its growth yield and was not able to use NH2OH as the sole N source. A non-haem hydroxylamine oxidoreductase was partially purified from A. faecalis strain TUD. The enzyme could only use K3Fe(CN)6 as an electron acceptor and reacted with antibodies raised against the hydroxylamine oxidoreductase of Thiosphaera pantotropha.
Otte
S
S
Kluyver laboratory for Biotechnology, Environmental Microbiology, Delft University of Technology, The Netherlands.
Schalk
J
J
Kuenen
J G
JG
Jetten
M S
MS
D000421:Alcaligenes / Q000201:enzymology / Q000254:growth & development / Q000378:metabolism*; D000641:Ammonia / Q000378:metabolism; D015153:Blotting, Western; D002849:Chromatography, Gas; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D019811:Hydroxylamine / Q000378:metabolism*; D013058:Mass Spectrometry; D009609:Nitrous Oxide / Q000378:metabolism*; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000302:isolation & purification / Q000378:metabolism
D003470:Culture Media; D019811:Hydroxylamine; D000641:Ammonia; D010088:Oxidoreductases; C021401:hydroxylamine dehydrogenase; D009609:Nitrous Oxide