Abstract
The nitrogen removal efficiency of heterotrophic nitrification and aerobic denitrification (HN–AD) bacteria can be seriously inhibited at low temperatures (< 15 °C). A novel psychrotolerant bacterium, Pseudomonas peli NR-5 (P. peli NR-5), with efficient HN–AD capability was isolated and screened from river sediments in cold areas. When P. peli NR-5 was aerobically cultivated for 60 h at 10 °C with NH4+-N, NO3−-N, and NO2−-N as the sole nitrogen sources (N 105 mg/L), the nitrogen removal efficiencies were 97.3, 95.3, and 87.8%, respectively, without nitrite accumulation, and the corresponding average nitrogen removal rates were 1.71, 1.67, and 1.55 mg/L/h, respectively. Meanwhile, P. peli NR-5 exhibited excellent simultaneous nitrification and denitrification capabilities at 10 °C. Sodium succinate was the most favorable carbon substrate for bacterial growth and ammonia removal by strain NR-5. The optimal culture conditions determined by the response surface methodology model were a carbon to nitrogen ratio of 5.9, temperature of 11.5 °C, pH of 7.0, and shaking speed of 144 rpm. Under these conditions, 99.1% of the total nitrogen was removed in the verification experiments, which was not significantly different from the predicted maximum removal in the model (99.6%). Six functional genes participating in the HN–AD process were successfully obtained by polymerase chain reaction amplification, which further confirmed the HN–AD capability of P. peli NR-5 and proposed the metabolic pathway of HN–AD. The above results provide a theoretical background of psychrotolerant HN–AD bacteria in wastewater purification under low-temperature conditions.