The development of an effective biotrickling filter (BF) system to inoculate a newly isolated strain of Pseudomonas putida SB1 for the effective treatment of nitric oxide (NO) is described. The experiments were carried out in a bench-scale BF under high concentrations of O 2 and NO in simulated flue gas. A method including alternating aeration in screening and rescreening based on the pH changes for cultivating natural aerobic denitrifying bacteria was employed. The SB1 showed a denitrifying capability of 95% nitrate removal rate over a 24 h period in an aerobic environment, with no nitrite accumulation. The BF system was able to consistently remove 82.9–94.2% NO when the inlet NO was 400 ppm in an enriched oxygen stream of 2–20%. The oxygen had no negative effect on the aerobic denitrifier SB1, but rather enhanced the total efficiency in part by chemical oxidation and in part by the strain activities. A kinetic relation between the oxygen concentration and biological NO removal was developed to confirm that the microbial metabolism played the main role. 79.3% of the total NO removal can be attributed to bio-denitrifying at 20% oxygen, and most chemical oxidation occurred concurrently. Overall, the study demonstrates that NO removal by the aerobic denitrifying process in BF is feasible in flue gas.