Nitrite (NO₂^-) can accumulate during nitrification in soil following fertilizer application. While the role of NO₂^- as a substrate regulating nitrous oxide (N₂O) production is recognized, kinetic data are not available that allow for estimating N₂O production or soil-to-atmosphere fluxes as a function of NO₂^- levels under aerobic conditions. The current study investigated these kinetics as influenced by soil physical and biochemical factors in soils from cultivated and uncultivated fields in Minnesota, USA. A linear response of N₂O production rate (P_N2O) to NO₂^- was observed at concentrations below 60 μg N g^-1 soil in both non-sterile and sterilized soils. Rate coefficients (K_p) relating P_N2O to NO₂^- varied over two orders of magnitude and were correlated with pH, total nitrogen, and soluble and total carbon (C). Total C explained 84% of the variance in K_p across all samples. Abiotic processes accounted for 31–75% of total N₂O production. Biological reduction of NO₂^- was enhanced as oxygen (O₂) levels were decreased from above ambient to 5%, consistent with nitrifier denitrification. In contrast, nitrate (NO₃^-)-reduction, and the reduction of N₂O itself, were only stimulated at O₂ levels below 5%. Greater temperature sensitivity was observed for biological compared to chemical N₂O production. Steady-state model simulations predict that NO₂^- levels often found after fertilizer applications have the potential to generate substantial N₂O fluxes even at ambient O₂. This potential derives in part from the production of N₂O under conditions not favorable for N₂O reduction, in contrast to N₂O generated from NO₃^- reduction. These results have implications with regard to improved management to minimize agricultural N₂O emissions and improved emissions assessments.