{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T07:06:13Z","timestamp":1789628773139,"version":"build-2803163510"},"reference-count":50,"publisher":"Oxford University Press (OUP)","issue":"1","license":[{"start":{"date-parts":[[2024,9,2]],"date-time":"2024-09-02T00:00:00Z","timestamp":1725235200000},"content-version":"vor","delay-in-days":245,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["52270026"],"award-info":[{"award-number":["52270026"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012226","name":"Fundamental Research Funds for the Central Universities","doi-asserted-by":"publisher","award":["DUT20RC(3)093"],"award-info":[{"award-number":["DUT20RC(3)093"]}],"id":[{"id":"10.13039\/501100012226","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Open Project of National & Local Joint Engineering Laboratory for Municipal Sewage Resource Utilization Technology","award":["2022KF04"],"award-info":[{"award-number":["2022KF04"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2024,1,8]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>The last two decades have seen nitrogen\/iron-transforming bacteria at the forefront of new biogeochemical discoveries, such as anaerobic ammonium oxidation coupled to ferric iron reduction (feammox) and lithoautotrophic nitrate-reducing ferrous iron-oxidation (NRFeOx). These emerging findings continue to expand our knowledge of the nitrogen\/iron cycle in nature and also highlight the need to re-understand the functional traits of the microorganisms involved. Here, as a proof-of-principle, we report compelling evidence for the capability of an NRFeOx enrichment culture to catalyze the feammox process. Our results demonstrate that the NRFeOx culture predominantly oxidizes NH4+ to nitrogen gas, by reducing both chelated nitrilotriacetic acid (NTA)-Fe(III) and poorly soluble Fe(III)-bearing minerals (\u03b3-FeOOH) at pH 4.0 and 8.0, respectively. In the NRFeOx culture, Fe(II)-oxidizing bacteria of Rhodanobacter and Fe(III)-reducing bacteria of unclassified_Acidobacteriota coexisted. Their relative abundances were dynamically regulated by the supplemented iron sources. Metagenomic analysis revealed that the NRFeOx culture contained a complete set of denitrifying genes along with hao genes for ammonium oxidation. Additionally, numerous genes encoding extracellular electron transport-associated proteins or their homologs were identified, which facilitated the reduction of extracellular iron by this culture. More broadly, this work lightens the unexplored potential of specific microbial groups in driving nitrogen transformation through multiple pathways and highlights the essential role of microbial iron metabolism in the integral biogeochemical nitrogen cycle.<\/jats:p>","DOI":"10.1093\/ismejo\/wrae149","type":"journal-article","created":{"date-parts":[[2024,8,1]],"date-time":"2024-08-01T03:17:15Z","timestamp":1722482235000},"source":"Crossref","is-referenced-by-count":16,"title":["Anaerobic ammonium oxidation coupled to iron(III) reduction catalyzed by a lithoautotrophic nitrate-reducing iron(II) oxidizing enrichment culture"],"prefix":"10.1093","volume":"18","author":[{"given":"Hong-Bin","family":"Zhang","sequence":"first","affiliation":[{"name":"Key Laboratory of Industrial Ecology and Environmental Engineering (Ministry of Education , China), School of Environmental Science and Technology, , Dalian 116024, China"},{"name":"Dalian University of Technology , China), School of Environmental Science and Technology, , Dalian 116024, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"He-Fei","family":"Wang","sequence":"additional","affiliation":[{"name":"National Marine Environmental Monitoring Center , Laboratory of Island Ecological Environment Protection, Dalian 116023, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jia-Bo","family":"Liu","sequence":"additional","affiliation":[{"name":"Key Laboratory of Industrial Ecology and Environmental Engineering (Ministry of Education , China), School of Environmental Science and Technology, , Dalian 116024, China"},{"name":"Dalian University of Technology , China), School of Environmental Science and Technology, , Dalian 116024, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhen","family":"Bi","sequence":"additional","affiliation":[{"name":"School of Environment Science and Engineering, Suzhou University of Science and Technology , Suzhou 215009, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruo-Fei","family":"Jin","sequence":"additional","affiliation":[{"name":"Key Laboratory of Industrial Ecology and Environmental Engineering (Ministry of Education , China), School of Environmental Science and Technology, , Dalian 116024, China"},{"name":"Dalian University of Technology , China), School of Environmental Science and Technology, , Dalian 116024, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tian","family":"Tian","sequence":"additional","affiliation":[{"name":"Key Laboratory of Industrial Ecology and Environmental Engineering (Ministry of Education , China), School of Environmental Science and Technology, , Dalian 116024, China"},{"name":"Dalian University of Technology , China), School of Environmental Science and Technology, , Dalian 116024, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2024,7,31]]},"reference":[{"key":"2024090207154921200_ref1","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1007\/BF00002772","article-title":"Nitrogen limitation on land and in the sea: how can it occur","volume":"13","author":"Vitousek","year":"1991","journal-title":"Biogeochemistry"},{"key":"2024090207154921200_ref2","doi-asserted-by":"crossref","first-page":"538","DOI":"10.1038\/ngeo1530","article-title":"Nitrogen loss from soil through anaerobic ammonium oxidation coupled to iron reduction","volume":"5","author":"Yang","year":"2012","journal-title":"Nat Geosci"},{"key":"2024090207154921200_ref3","doi-asserted-by":"crossref","first-page":"217","DOI":"10.1038\/ngeo2886","article-title":"Iron-dependent nitrogen cycling in a ferruginous lake and the nutrient status of Proterozoic oceans","volume":"10","author":"Michiels","year":"2017","journal-title":"Nat Geosci"},{"key":"2024090207154921200_ref4","doi-asserted-by":"crossref","first-page":"9298","DOI":"10.1021\/acs.est.6b02077","article-title":"Electron shuttles enhance anaerobic ammonium oxidation coupled to iron(III) reduction","volume":"50","author":"Zhou","year":"2016","journal-title":"Environ Sci Technol"},{"key":"2024090207154921200_ref5","doi-asserted-by":"crossref","first-page":"3357","DOI":"10.1038\/s41396-021-01004-x","article-title":"In-depth analysis of N2O fluxes in tropical forest soils of the Congo Basin combining isotope and functional gene analysis","volume":"15","author":"Gallarotti","year":"2021","journal-title":"ISME J"},{"key":"2024090207154921200_ref6","doi-asserted-by":"crossref","first-page":"277","DOI":"10.1111\/gbi.12540","article-title":"Iron-mediated anaerobic ammonium oxidation recorded in the early Archean ferruginous ocean","volume":"21","author":"Pellerin","year":"2023","journal-title":"Geobiology"},{"key":"2024090207154921200_ref7","doi-asserted-by":"crossref","DOI":"10.1016\/j.watres.2022.118867","article-title":"Feammox in alluvial-lacustrine aquifer system: nitrogen\/iron isotopic and biogeochemical evidences","volume":"222","author":"Xiong","year":"2022","journal-title":"Water Res"},{"key":"2024090207154921200_ref8","doi-asserted-by":"crossref","first-page":"11560","DOI":"10.1021\/acs.est.5b03419","article-title":"Evidence of nitrogen loss from anaerobic ammonium oxidation coupled with ferric iron reduction in an intertidal wetland","volume":"49","author":"Li","year":"2015","journal-title":"Environ Sci Technol"},{"key":"2024090207154921200_ref9","doi-asserted-by":"crossref","first-page":"10641","DOI":"10.1021\/es503113s","article-title":"Nitrogen loss through anaerobic ammonium oxidation coupled to iron reduction from paddy soils in a chronosequence","volume":"48","author":"Ding","year":"2014","journal-title":"Environ Sci Technol"},{"key":"2024090207154921200_ref10","doi-asserted-by":"crossref","first-page":"4841","DOI":"10.1021\/acs.est.8b05016","article-title":"RNA stable isotope probing of potential Feammox population in paddy soil","volume":"53","author":"Li","year":"2019","journal-title":"Environ Sci Technol"},{"key":"2024090207154921200_ref11","doi-asserted-by":"crossref","first-page":"3462","DOI":"10.1111\/1462-2920.14328","article-title":"Microbial anaerobic Fe(II) oxidation \u2013 ecology, mechanisms and environmental implications","volume":"20","author":"Bryce","year":"2018","journal-title":"Environ Microbiol"},{"key":"2024090207154921200_ref12","doi-asserted-by":"crossref","DOI":"10.1016\/j.biotechadv.2020.107610","article-title":"Iron-assisted biological wastewater treatment: synergistic effect between iron and microbes","volume":"44","author":"Tian","year":"2020","journal-title":"Biotechnol Adv"},{"key":"2024090207154921200_ref13","doi-asserted-by":"crossref","first-page":"1458","DOI":"10.1128\/aem.62.4.1458-1460.1996","article-title":"Anaerobic, nitrate-dependent microbial oxidation of ferrous iron","volume":"62","author":"Straub","year":"1996","journal-title":"Appl Environ Microbiol"},{"key":"2024090207154921200_ref14","doi-asserted-by":"crossref","DOI":"10.1016\/j.watres.2019.115300","article-title":"Exclusive microbially driven autotrophic iron-dependent denitrification in a reactor inoculated with activated sludge","volume":"170","author":"Tian","year":"2020","journal-title":"Water Res"},{"key":"2024090207154921200_ref15","doi-asserted-by":"crossref","first-page":"AEM00460","DOI":"10.1128\/AEM.00460-21","article-title":"Nitrate removal by a novel lithoautotrophic nitrate-reducing, iron-reducing, iron(II)-oxidizing culture enriched from a pyrite-rich limestone aquifer","volume":"87","author":"Jakus","year":"2021","journal-title":"Appl Environ Microbiol"},{"key":"2024090207154921200_ref16","doi-asserted-by":"crossref","first-page":"280","DOI":"10.1159\/000517083","article-title":"A novel enrichment culture highlights core features of microbial networks contributing to autotrophic Fe(II) oxidation coupled to nitrate reduction","volume":"31","author":"Huang","year":"2021","journal-title":"Microb Physiol"},{"key":"2024090207154921200_ref17","first-page":"360","article-title":"An evolving view on biogeochemical cycling of iron","volume":"19","author":"Kappler","year":"2021","journal-title":"Nature"},{"key":"2024090207154921200_ref18","doi-asserted-by":"crossref","first-page":"e00752","DOI":"10.1128\/AEM.00752-17","article-title":"Insights into nitrate-reducing Fe(II) oxidation mechanisms through analysis of cell-mineral associations, cell encrustation, and mineralogy in the chemolithoautotrophic enrichment culture KS","volume":"83","author":"Nordhoff","year":"2017","journal-title":"Appl Environ Microbiol"},{"key":"2024090207154921200_ref19","doi-asserted-by":"crossref","first-page":"10646","DOI":"10.1021\/acs.est.0c02521","article-title":"Electron hopping enables rapid electron transfer between quinone\u2212\/hydroquinone-containing organic molecules in microbial iron(III) mineral reduction","volume":"54","author":"Bai","year":"2020","journal-title":"Environ Sci Technol"},{"key":"2024090207154921200_ref20","article-title":"Specifically adsorbed ferrous ions modulate interfacial affinity for high-rate ammonia electrosynthesis from nitrate in neutral media","volume":"120","author":"Liu","year":"2023","journal-title":"Proc Natl Acad Sci USA"},{"key":"2024090207154921200_ref21","doi-asserted-by":"crossref","first-page":"345","DOI":"10.1016\/j.cej.2018.03.156","article-title":"Chemolithotrophic denitrification by nitrate-dependent anaerobic iron oxidizing (NAIO) process: insights into the evaluation of seeding sludge","volume":"345","author":"Zhang","year":"2018","journal-title":"Chem Eng J"},{"key":"2024090207154921200_ref22","volume-title":"Standard Methods for the Examination of Water and Wastewater, American Water Works Association, Twenty-First Ed.","author":"A.P.H.A","year":"2005"},{"key":"2024090207154921200_ref23","doi-asserted-by":"crossref","first-page":"180","DOI":"10.1111\/gbi.12019","article-title":"Abiotic oxidation of Fe(II) by reactive nitrogen species in cultures of the nitrate-reducing Fe(II) oxidizer Acidovorax sp. BoFeN1 - questioning the existence of enzymatic Fe(II) oxidation","volume":"11","author":"Klueglein","year":"2013","journal-title":"Geobiology"},{"key":"2024090207154921200_ref24","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1080\/01490451.2017.1303554","article-title":"A revised iron extraction protocol for environmental samples rich in nitrite and carbonate","volume":"35","author":"Schaedler","year":"2018","journal-title":"Geomicrobiol J"},{"key":"2024090207154921200_ref25","doi-asserted-by":"crossref","first-page":"11576","DOI":"10.1021\/acs.est.0c02882","article-title":"Phosphorus recovery from wastewater prominently through a Fe(II)-P oxidizing pathway in the autotrophic iron-dependent denitrification process","volume":"54","author":"Tian","year":"2020","journal-title":"Environ Sci Technol"},{"key":"2024090207154921200_ref26","doi-asserted-by":"crossref","first-page":"6120","DOI":"10.1128\/AEM.01570-16","article-title":"Evidence for the existence of autotrophic nitrate-reducing Fe(II)-oxidizing bacteria in marine coastal sediment","volume":"82","author":"Laufer","year":"2016","journal-title":"Appl Environ Microbiol"},{"key":"2024090207154921200_ref27","doi-asserted-by":"crossref","first-page":"6449","DOI":"10.1021\/acs.est.6b00562","article-title":"Stimulation of Fe(II) oxidation, biogenic lepidocrocite formation, and arsenic immobilization by Pseudogulbenkiania sp. strain 2002","volume":"50","author":"Xiu","year":"2016","journal-title":"Environ Sci Technol"},{"key":"2024090207154921200_ref28","doi-asserted-by":"crossref","first-page":"11410","DOI":"10.1021\/acs.est.9b04047","article-title":"Defluorination of perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS) by Acidimicrobium sp. strain A6","volume":"53","author":"Huang","year":"2019","journal-title":"Environ Sci Technol"},{"key":"2024090207154921200_ref29","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1016\/j.soilbio.2016.04.012","article-title":"Environmental factors affecting the presence of Acidimicrobiaceae and ammonium removal under iron-reducing conditions in soil environments","volume":"98","author":"Huang","year":"2016","journal-title":"Soil Biol Biochem"},{"key":"2024090207154921200_ref30","doi-asserted-by":"crossref","first-page":"5255","DOI":"10.1128\/AEM.69.9.5255-5262.2003","article-title":"Nitrilotriacetate stimulation of anaerobic Fe(III) respiration by mobilization of humic materials in soil","volume":"69","author":"Luu","year":"2003","journal-title":"Appl Environ Microbiol"},{"key":"2024090207154921200_ref31","doi-asserted-by":"crossref","first-page":"4131","DOI":"10.1021\/acs.est.9b07134","article-title":"AQDS and redox-active NOM enables microbial Fe(III)-mineral reduction at cm-scales","volume":"54","author":"Bai","year":"2020","journal-title":"Environ Sci Technol"},{"key":"2024090207154921200_ref32","doi-asserted-by":"crossref","first-page":"8161","DOI":"10.1021\/acs.chemrev.0c01286","article-title":"Fe(II) redox chemistry in the environment","volume":"121","author":"Huang","year":"2021","journal-title":"Chem Rev"},{"key":"2024090207154921200_ref33","doi-asserted-by":"crossref","first-page":"e02173","DOI":"10.1128\/AEM.02173-17","article-title":"Growth and population dynamics of the anaerobic Fe(II)-oxidizing and nitrate-reducing enrichment culture KS","volume":"84","author":"Tominski","year":"2018","journal-title":"Appl Environ Microbiol"},{"key":"2024090207154921200_ref34","doi-asserted-by":"crossref","first-page":"32617","DOI":"10.1073\/pnas.2005917117","article-title":"Geochemical transition zone powering microbial growth in subsurface sediments","volume":"117","author":"Zhao","year":"2020","journal-title":"Proc Natl Acad Sci USA"},{"key":"2024090207154921200_ref35","doi-asserted-by":"crossref","first-page":"e02166","DOI":"10.1128\/AEM.02166-17","article-title":"Insights into carbon metabolism provided by fluorescence in situ hybridization-secondary ion mass spectrometry imaging of an autotrophic, nitrate-reducing, Fe(II)-oxidizing enrichment culture","volume":"84","author":"Tominski","year":"2018","journal-title":"Appl Environ Microbiol"},{"key":"2024090207154921200_ref36","doi-asserted-by":"crossref","first-page":"e00496","DOI":"10.1128\/AEM.00496-21","article-title":"Meta-omics reveal Gallionellaceae and Rhodanobacter species as interdependent key players for Fe(II) oxidation and nitrate reduction in the autotrophic enrichment culture KS","volume":"87","author":"Huang","year":"2021","journal-title":"Appl Environ Microbiol"},{"key":"2024090207154921200_ref37","doi-asserted-by":"crossref","first-page":"1231","DOI":"10.1038\/ismej.2008.75","article-title":"Bacterial associated with iron seeps in a sulfur-rich, neutral pH, freshwater ecosystem","volume":"2","author":"Haaijer","year":"2008","journal-title":"ISME J"},{"key":"2024090207154921200_ref38","doi-asserted-by":"crossref","first-page":"752","DOI":"10.1038\/nrmicro1490","article-title":"Microorganisms pumping iron: anaerobic microbial iron oxidation and reduction","volume":"4","author":"Webber","year":"2006","journal-title":"Nat Rev Microbiol"},{"key":"2024090207154921200_ref39","article-title":"Isolation and characterization of an ammonium-oxidizing iron reducer: Acidimicrobiaceae sp. A6","volume":"13","author":"Huang","year":"2018","journal-title":"PLoS One"},{"key":"2024090207154921200_ref40","doi-asserted-by":"crossref","first-page":"2058","DOI":"10.1038\/s41467-020-16016-y","article-title":"Extracellular electron transfer-dependent anaerobic oxidation of ammonium by anammox bacteria","volume":"11","author":"Shaw","year":"2020","journal-title":"Nat Commun"},{"key":"2024090207154921200_ref41","doi-asserted-by":"crossref","first-page":"168","DOI":"10.1016\/j.watres.2017.11.059","article-title":"Microbial electricity driven anoxic ammonium removal","volume":"130","author":"Vilajeliu-Pons","year":"2018","journal-title":"Water Res"},{"key":"2024090207154921200_ref42","doi-asserted-by":"crossref","first-page":"12883","DOI":"10.1073\/pnas.1410551111","article-title":"Shewanella oneidensis MR-1 nanowires are outer membrane and periplasmic extensions of the ectracellular electron transport components","volume":"111","author":"Pirbadian","year":"2014","journal-title":"Proc Natl Acad Sci USA"},{"key":"2024090207154921200_ref43","doi-asserted-by":"crossref","first-page":"5082","DOI":"10.1021\/acs.est.6b04640","article-title":"Enhancing extracellular electron transfer of Shewanella oneidensis MR-1 through coupling improved flavin synthesis and metal-reducing conduit for pollutant degradation","volume":"51","author":"Min","year":"2017","journal-title":"Environ Sci Technol"},{"key":"2024090207154921200_ref44","doi-asserted-by":"crossref","first-page":"2656","DOI":"10.1128\/AEM.03493-15","article-title":"Metagenomic analyses of the autotrophic Fe(II)-oxidizing, nitrate-reducing enrichment culture KS","volume":"82","author":"He","year":"2016","journal-title":"Appl Environ Microbiol"},{"key":"2024090207154921200_ref45","doi-asserted-by":"crossref","first-page":"25803","DOI":"10.1074\/jbc.M802496200","article-title":"A new iron-oxidizing\/O2-reducing supercomplex spanning both inner and outer membranes, isolated from the extreme acidophile Acidithiobacillus ferrooxidans","volume":"283","author":"Castelle","year":"2008","journal-title":"J Biol Chem"},{"key":"2024090207154921200_ref46","doi-asserted-by":"crossref","first-page":"453","DOI":"10.1038\/s41586-020-2468-5","article-title":"Bacterial chemolithoautotrophy via manganese oxidation","volume":"583","author":"Yu","year":"2020","journal-title":"Nature"},{"key":"2024090207154921200_ref47","first-page":"E92","article-title":"Metagenomics-guided analysis of microbial chemolithoautotrophic phosphite oxidation yields evidence of a seventh natural CO2 fixation pathway","volume":"115","author":"Figueroa","year":"2017","journal-title":"Proc Natl Acad Sci USA"},{"key":"2024090207154921200_ref48","doi-asserted-by":"crossref","first-page":"E9271","DOI":"10.1073\/pnas.1810386115","article-title":"Assimilation of formic acid and CO2 by engineered Escherichia coli equipped with reconstructed one-carbon assimilation pathways","volume":"115","author":"Bang","year":"2018","journal-title":"Proc Natl Acad Sci USA"},{"key":"2024090207154921200_ref49","doi-asserted-by":"crossref","first-page":"7852","DOI":"10.1039\/D0GC03493B","article-title":"Biological carbon fixation: a thermodynamic perspective","volume":"23","author":"Zhao","year":"2021","journal-title":"Green Chem"},{"key":"2024090207154921200_ref50","doi-asserted-by":"crossref","DOI":"10.1073\/pnas.2007051117","article-title":"Lithogenic hydrogen supports microbial primary production in subglacial and proglacial environments","volume":"118","author":"Dunham","year":"2021","journal-title":"Proc Natl Acad Sci USA"}],"container-title":["The ISME Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/ismej\/advance-article-pdf\/doi\/10.1093\/ismejo\/wrae149\/58704018\/wrae149.pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/ismej\/article-pdf\/18\/1\/wrae149\/58991296\/wrae149.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/ismej\/article-pdf\/18\/1\/wrae149\/58991296\/wrae149.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,9,2]],"date-time":"2024-09-02T07:44:07Z","timestamp":1725263047000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/ismej\/article\/doi\/10.1093\/ismejo\/wrae149\/7724874"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,1]]},"references-count":50,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2024,1,8]]}},"URL":"https:\/\/doi.org\/10.1093\/ismejo\/wrae149","relation":{},"ISSN":["1751-7362","1751-7370"],"issn-type":[{"value":"1751-7362","type":"print"},{"value":"1751-7370","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2024,1]]},"published":{"date-parts":[[2024,1]]},"article-number":"wrae149"}}