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Peer-reviewedCC BY 4.0mBio

Diversity of dirammox microorganisms

1 more authors
4 affiliations
  1. 1State Key Laboratory of Microbial Diversity and Innovative Utilization, Institute of Microbiology, Chinese Academy of Sciences
  2. 2College of Life Sciences, University of Chinese Academy of Sciences
  3. 3China General Microbiological Culture Collection Center, Institute of Microbiology, Chinese Academy of Sciences
  4. 4Key Laboratory of Agricultural and Environmental Microbiology, Ministry of Agriculture and Rural Affairs, College of Life Sciences, Nanjing Agricultural University

Abstract

Our knowledge of the nitrogen cycle has been advanced by the discovery of anammox bacteria, ammonia-oxidizing archaea, and comammox bacteria, along with their high diversity. Direct ammonia oxidation (dirammox), a recently discovered heterotrophic nitrification pathway, converts ammonium and organic nitrogen directly to N ₂ through the DnfABC multienzyme system, further expanding our understanding of the nitrogen cycle. However, the taxonomic distribution, prevalence, and nitrogen metabolic potential of microorganisms encoding dirammox genes remain poorly understood. Here, large-scale genome mining identified complete dnfABC gene clusters in 690 genomes representing 3 phyla and 26 genera. The dnfABC cluster showed a striking bimodal prevalence: it was nearly ubiquitous in Alcaligenes , Delftia , and Microvirgula but occurred sporadically in most other genera. These microorganisms were recovered from diverse environments, particularly organic nitrogen-rich habitats. Comparative analyses of DnfABC phylogeny, gene cluster organization, GC content, and flanking mobile genetic elements indicate that the dnfABC cluster is conserved within several core genera, whereas its sporadic distribution in other lineages suggests that horizontal gene transfer may have contributed to its dissemination. The dnfABC cluster co-occurs with genes associated with assimilatory nitrate/nitrite reduction to ammonium, dissimilatory nitrate/nitrite reduction to ammonium, denitrification, and/or nitrogen fixation, but not with marker genes for nitrification or anammox, indicating diverse nitrogen metabolic potential among potential dirammox microorganisms. Functional assays further showed that diverse dnfABC clusters consistently conferred ammonium- and glutamine-oxidizing activities in Escherichia coli , despite exhibiting low activities in their native hosts. These results suggest that dirammox is a broadly distributed, metabolically flexible, and tightly regulated pathway and may represent an overlooked route of nitrogen transformation in organic-rich habitats.

IMPORTANCE: Nitrogen is essential for all life, and our understanding of the nitrogen cycle has advanced substantially with the discovery of anammox bacteria, ammonia-oxidizing archaea, and comammox bacteria, which exhibit remarkable taxonomic and metabolic diversity. This study suggests that the recently discovered dirammox pathway may be more widespread in nature than previously recognized. Potential dirammox microorganisms exhibit broad phylogenetic and ecological distributions, high genomic diversity, and notable metabolic versatility. Functional assays indicate that dirammox activity is tightly regulated in native hosts, suggesting that its ecological role depends on specific environmental conditions. Unlike canonical nitrification and anammox, dirammox operates under conditions where other nitrogen transformations are limited, highlighting an important route of nitrogen loss in organic-rich habitats. These findings highlight the previously overlooked potential role of dirammox in nitrogen cycling, expand our understanding of microbial contributions to the global nitrogen cycle, and provide a foundation for future ecological and environmental research.

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BibTeX
@article{khasbaatar2026diversity,
  title = {{Diversity of dirammox microorganisms}},
  author = {Gantuya Khasbaatar and Qun Han and Xiao-Kang Wang and Lei Song and Chang Liu and Xin Sun and Ze-Shen Liu and Ying Liu and Cheng-Ying Jiang and Ji-Guo Qiu and Shuang-Jiang Liu and Yanning Zheng and De-Feng Li},
  journal = {mBio},
  year = {2026},
  month = oct,
  doi = {10.1128/mbio.01645-26},
  url = {https://journals.asm.org/doi/10.1128/mbio.01645-26},
  note = {CC BY 4.0},
}
RIS
TY  - JOUR
TI  - Diversity of dirammox microorganisms
AU  - Gantuya Khasbaatar
AU  - Qun Han
AU  - Xiao-Kang Wang
AU  - Lei Song
AU  - Chang Liu
AU  - Xin Sun
AU  - Ze-Shen Liu
AU  - Ying Liu
AU  - Cheng-Ying Jiang
AU  - Ji-Guo Qiu
AU  - Shuang-Jiang Liu
AU  - Yanning Zheng
AU  - De-Feng Li
PY  - 2026
DA  - 2026/10/15
JO  - mBio
DO  - 10.1128/mbio.01645-26
UR  - https://journals.asm.org/doi/10.1128/mbio.01645-26
AB  - Our knowledge of the nitrogen cycle has been advanced by the discovery of anammox bacteria, ammonia-oxidizing archaea, and comammox bacteria, along with their high diversity. Direct ammonia oxidation (dirammox), a recently discovered heterotrophic nitrification pathway, converts ammonium and organic nitrogen directly to N ₂ through the DnfABC multienzyme system, further expanding our understanding of the nitrogen cycle. However, the taxonomic distribution, prevalence, and nitrogen metabolic potential of microorganisms encoding dirammox genes remain poorly understood. Here, large-scale genome mining identified complete dnfABC gene clusters in 690 genomes representing 3 phyla and 26 genera. The dnfABC cluster showed a striking bimodal prevalence: it was nearly ubiquitous in Alcaligenes , Delftia , and Microvirgula but occurred sporadically in most other genera. These microorganisms were recovered from diverse environments, particularly organic nitrogen-rich habitats. Comparative analyses of DnfABC phylogeny, gene cluster organization, GC content, and flanking mobile genetic elements indicate that the dnfABC cluster is conserved within several core genera, whereas its sporadic distribution in other lineages suggests that horizontal gene transfer may have contributed to its dissemination. The dnfABC cluster co-occurs with genes associated with assimilatory nitrate/nitrite reduction to ammonium, dissimilatory nitrate/nitrite reduction to ammonium, denitrification, and/or nitrogen fixation, but not with marker genes for nitrification or anammox, indicating diverse nitrogen metabolic potential among potential dirammox microorganisms. Functional assays further showed that diverse dnfABC clusters consistently conferred ammonium- and glutamine-oxidizing activities in Escherichia coli , despite exhibiting low activities in their native hosts. These results suggest that dirammox is a broadly distributed, metabolically flexible, and tightly regulated pathway and may represent an overlooked route of nitrogen transformation in organic-rich habitats.  IMPORTANCE: Nitrogen is essential for all life, and our understanding of the nitrogen cycle has advanced substantially with the discovery of anammox bacteria, ammonia-oxidizing archaea, and comammox bacteria, which exhibit remarkable taxonomic and metabolic diversity. This study suggests that the recently discovered dirammox pathway may be more widespread in nature than previously recognized. Potential dirammox microorganisms exhibit broad phylogenetic and ecological distributions, high genomic diversity, and notable metabolic versatility. Functional assays indicate that dirammox activity is tightly regulated in native hosts, suggesting that its ecological role depends on specific environmental conditions. Unlike canonical nitrification and anammox, dirammox operates under conditions where other nitrogen transformations are limited, highlighting an important route of nitrogen loss in organic-rich habitats. These findings highlight the previously overlooked potential role of dirammox in nitrogen cycling, expand our understanding of microbial contributions to the global nitrogen cycle, and provide a foundation for future ecological and environmental research.
SN  - 2150-7511
ER  -