Microbial manganese redox cycling drives co-removal of nitrate and ammonium

Manganese (Mn), abundant in the Earth's crust, can act as an oxidant or a reductant for diverse nitrogen biotransformation processes. However, the functional microorganisms and their metabolic pathways, as well as interactions, remain largely elusive. Here, a microbial consortium was enriched from a mixture of freshwater sediments and activated sludge by feeding ammonium, nitrate and Mn(II), which established manganese-driven co-removal of nitrate and ammonium with removal rates of 5.83 and 2.30 mg N L⁻¹ d⁻¹, respectively. The batch tests and metagenomic analyses revealed a nitrate-dependent anaerobic manganese oxidation (NDMO) process mediated by Anaerolineales and Phycisphaerales and a manganese reduction coupled to anaerobic ammonium oxidation (Mnammox) process mediated by Chthonomonadales. Based on identified key genes involved in the nitrogen and manganese metabolic pathways, nitrate was likely reduced to nitrite and nitrogen gas in the NDMO process while ammonium was oxidized to nitrite in the Mnammox process, which in turn fuelled the Anammox process carried out by Candidatus Brocadia. This revealed the microbial interactions of NDMO, Mnammox, and Anammox processes responsible for manganese-driven co-removal of ammonium and nitrate. These findings provide a potential solution for biological nitrogen removal and expand our understanding of the nitrogen and manganese biogeochemical cycles.