Clarifying Microbial Nitrous Oxide Reduction Under Aerobic Conditions: Tolerant, Intolerant, and Sensitive

Abstract

ABSTRACTOne of the major challenges for the bioremediation application of microbial N2O reduction is its oxygen sensitivity. While a few strains were reported capable of reducing N2O under aerobic conditions, the N2O reduction kinetics of phylogenetically diverse N2O reducers are not well understood. Here we analyzed and compared the kinetics of Clade I and Clade II N2O-reducing bacteria in the presence or absence of O2by using a whole-cell assay with N2O and O2microsensors. Among the seven strains tested, N2O reduction ofStutzerimonas stutzeriTR2 and ZoBell were not inhibited by oxygen (i.e., oxygen tolerant).Paracoccus denitrificans, Azospirillum brasilense, andGemmatimonas aurantiacareduced N2O in the presence of O2but slower than in the absence of O2(i.e., oxygen sensitive). N2O reduction ofPseudomonas aeruginosaandDechloromonas aromaticadid not occur when O2was present (i.e., oxygen intolerant). Amino acid sequences and predicted structures of NosZ were highly similar among these strains, whereas oxygen-tolerant N2O reducers had higher oxygen consumption rates. The results suggest that the mechanism of O2tolerance is not directly related to NosZ structure but rather related to the scavenging of O2in the cells and/or accessory proteins encoded by thenoscluster.IMPORTANCESome bacteria can reduce N2O in the presence of O2, whereas others cannot. It is unclear whether this trait of aerobic N2O reduction is related to the phylogeny and structure of N2O reductase. The understanding of aerobic N2O reduction is critical in guiding emission control, due to the common concurrence of N2O and O2in natural and engineered systems. This study provided the N2O reduction kinetics of various bacteria under aerobic and anaerobic conditions and classified the bacteria into oxygen-tolerant, -sensitive, and -intolerant N2O reducers. Oxygen-tolerant N2O reducers rapidly consumed O2, which could help maintain the low O2concentration in the cells and keep their N2O reductase active. These findings are important and useful when selecting N2O reducers for bioremediation applications.