Pathways of N2O production by marine ammonia-oxidizing archaea determined from dual-isotope labeling

Xianhui S. Wan, Lei Hou, Shuh Ji Kao, Yao Zhang, Hua Xia Sheng, Hui Shen, Senwei Tong, Wei Qin, Bess B. Ward

Research output: Contribution to journalArticlepeer-review

10 Scopus citations


The ocean is a net source of the greenhouse gas and ozone-depleting substance, nitrous oxide (N2O), to the atmosphere. Most of that N2O is produced as a trace side product during ammonia oxidation, primarily by ammonia-oxidizing archaea (AOA), which numerically dominate the ammonia-oxidizing community in most marine environments. The pathways to N2O production and their kinetics, however, are not completely understood. Here, we use 15N and 18O isotopes to determine the kinetics of N2O production and trace the source of nitrogen (N) and oxygen (O) atoms in N2O produced by a model marine AOA species, Nitrosopumilus maritimus. We find that during ammonia oxidation, the apparent half saturation constants of nitrite and N2O production are comparable, suggesting that both processes are enzymatically controlled and tightly coupled at low ammonia concentrations. The constituent atoms in N2O are derived from ammonia, nitrite, O2, and H2O via multiple pathways. Ammonia is the primary source of N atoms in N2O, but its contribution varies with ammonia to nitrite ratio. The ratio of 45N2O to 46N2O (i.e., single or double labeled N) varies with substrate ratio, leading to widely varying isotopic signatures in the N2O pool. O2 is the primary source for O atoms. In addition to the previously demonstrated hybrid formation pathway, we found a substantial contribution by hydroxylamine oxidation, while nitrite reduction is an insignificant source of N2O. Our study highlights the power of dual 15N-18O isotope labeling to disentangle N2O production pathways in microbes, with implications for interpretation of pathways and regulation of marine N2O sources.

Original languageEnglish (US)
Article numbere2220697120
JournalProceedings of the National Academy of Sciences of the United States of America
Issue number11
StatePublished - Mar 8 2023

All Science Journal Classification (ASJC) codes

  • General


  • ammonia-oxidizing archaea
  • dual isotope
  • kinetics
  • marine N2O production pathways
  • nitrous oxide


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