Stoichiometric and kinetic characterization of an acid-tolerant ammonia oxidizer ‘Candidatus Nitrosoglobus’

Recently, acidic (i.e. pH<5) nitrification in activated-sludge is attracting attention because it enables stable nitritation (NH₄⁺ → NO₂⁻), and enhances sludge reduction and stabilization. However, the key acid-tolerant ammonia oxidizers involved are poorly understood. In this study, we performed stoichiometric and kinetic characterization of a new acid-tolerant ammonia-oxidizing bacterium (AOB) belonging to gamma-proteobacterium, Candidatus Nitrosoglobus. Ca. Nitrosoglobus was cultivated in activated-sludge in a laboratory membrane bioreactor over 200 days, with a relative abundance of 55.1 ± 0.5% (indicated by 16S rRNA gene amplicon sequencing) at the time of the characterization experiments. Among all known nitrifiers, Ca. Nitrosoglobus bears the highest resistance to nitrite, low pH, and free nitrous acid (FNA). These traits define Ca. Nitrosoglobus as an adversity-strategist that tends to prosper in acidic activated-sludge, where the low pH (< 5.0) and high levels of FNA (at parts per million levels) sustained and inhibited all other nitrifiers. In contrast, in the conventional pH-neutral activated-sludge process, Ca. Nitrosoglobus is less competitive with canonical AOB (e.g. Nitrosomonas) due to the relatively slow specific growth rate and low affinities to both oxygen and total ammonia. These results advance our understanding of acid-tolerant ammonia oxidizers, and support further development of the acidic activated-sludge process in which Ca. Nitrosoglobus can play a critical role.