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Siderophores as a selective regulator for enhancing anaerobic ammonium oxidation bacteria

Nature Water volume 3, pages 806–817 (2025)Cite this article

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Abstract

The limited iron uptake efficiency of anaerobic ammonium oxidation (anammox) bacteria has garnered accumulating concerns, as iron plays an important role in anammox bacteria metabolism. Siderophores are a prevalent strategy for iron acquisition among many bacteria, and recent evidence has suggested the potential role of siderophore-Fe3+ as bioavailable iron sources to anammox bacteria. Nonetheless, what target siderophore anammox bacteria can use and how they use it remain poorly understood. Here we shed fresh light on siderophore selectively promoting iron uptake and regulating the metabolic activity of anammox bacteria. Batch experiments and long-term operations demonstrated that siderophores catechin (CAT) and N-hydroxyethyl ethylenediamine triacetic acid (HEDTA) increased the iron uptake efficiency of anammox consortia up to 50%–65% and triggered the cofactors synthesis for key enzymes assembly, thereby selectively enhancing the anammox activity (≥350 mg N per gram of volatile suspended solids per day) and nitrogen removal efficiency (≥85%). Integrated with multi-omics approaches, we revealed versatile iron uptake pathways within anammox bacteria genera: Candidatus Brocadia acquired CAT-Fe3+ and HEDTA-Fe3+ via outer membrane receptors FitA/TbpA and FecA, respectively, whereas Candidatus Jettenia initially reduced CAT-Fe3+ using NfnB enzyme before uptake via the FeoABC system. This work reveals a pivotal role of siderophores in selectively enhancing anammox bacteria by optimizing their iron uptake and utilization and uncovers the involved molecular mechanisms, which opens promising avenues to optimize the anammox process with siderophore regulation for more efficient and sustainable wastewater treatment.

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Fig. 1: Short-term effect of exogenous siderophores on anammox systems.
Fig. 2: Long-term operation performance of the anammox systems with different iron additions.
Fig. 3: Iron content and morphology of anammox sludge in the anammox systems.
Fig. 4: Characterization of anammox sludge morphology in the anammox systems at day 60.
Fig. 5: Functional genus and nitrogen metabolic level of anammox bacteria.
Fig. 6: Metagenomic and metatranscriptomic information related to iron metabolism in the anammox systems.
Fig. 7: Proposed mechanisms of iron uptake and nitrogen transformation of anammox bacteria mediated by siderophores.

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Data availability

The data generated in this study are included within the article and its Supplementary Information. Raw sequencing data have been archived in NCBI Sequence Read Archive under the accession number PRJNA1222017. Source data are provided with this paper.

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Acknowledgements

This work was supported by the National Science Fund for Distinguished Young Scholars (52225001).

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Authors and Affiliations

  1. State Key Laboratory of Water Pollution Control and Green Resource Recycling, Shanghai Institute of Pollution Control and Ecological Security, College of Environmental Science and Engineering, Tongji University, Shanghai, People’s Republic of China

    Jiawei Liu, Jia Li, Han Wang, Mingda Zhou & Yayi Wang

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  1. Jiawei Liu
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  2. Jia Li
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  3. Han Wang
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Contributions

Y.W. conceived and led the project. J. Liu conceived and conducted the experiment, developed the figures and wrote the paper. H.W. analysed the microbial and statistical data and assisted with writing the paper. J. Li and M.Z. assisted with the analysis of the experimental data. Y.W., H.W. and J. Liu commented upon and revised the manuscript. All authors discussed and interpreted the results and contributed to the manuscript.

Corresponding authors

Correspondence to Han Wang or Yayi Wang.

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The authors declare no competing interests.

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Peer review information

Nature Water thanks the anonymous reviewers for their contribution to the peer review of this work.

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Extended data

Extended Data Table 1 Long-term operational strategies of anammox reactors
Full size table
Extended Data Table 2 The relative content of elements in anammox sludge at day 60 according to EDS analysis
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Supplementary information

Supplementary Information

Supplementary Texts 1–7, Figs. 1–7 and Tables 1–5.

Reporting Summary

Source data

Source Data Figs. 1–3, 5 and 6

Statistical source data for Figs. 1–3, 5 and 6.

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Liu, J., Li, J., Wang, H. et al. Siderophores as a selective regulator for enhancing anaerobic ammonium oxidation bacteria. Nat Water 3, 806–817 (2025). https://doi.org/10.1038/s44221-025-00459-y

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