Nitrosomonas europaea, Nitrobacter winogradskyi and Alcaligenes faecalis—typical ammonia-oxidizer, nitrite-oxidizer and heterotrophic nitrifier were immobilized in PVA gel and employed in the study. Continuous experiments were conducted in their pure and mixed cultures with DO concentrations in the cultures kept at 4, 2, 0.5 mg·l−1. Comparisons among N2O emission from different cultures were made to show their N2O producing capabilities. Results showed that: compared with N. europaea and A. faecalis, N. winogradskyi produced negligible N2O. On the other hand, N. europaea had the highest N2O producing ability. Unit N. europaea produced N2O 18–53 times higher than unit A. faecalis did. However, due to the higher population of A. faecalis, N2O production of the A. faecalis culture was higher than that of the N. europaea culture when DO concentration in the cultures was 2 mg·l−1; whereas, N2O yields of the A. faecalis culture were smaller than those of the N. europaea culture at DO concentrations of 4 and 0.5 mg·l−1. N2O emitted from most the mixed cultures was lower than that from pure cultures under the experimental conditions.