To improve the biological nitrogen removal efficiency of petrochemical wastewater, three aerobic denitrifiers were isolated from caprolactam sewage treatment pool. They were identified as Acinetobacter sp. YY1, Sphingomonas sp. YY2 and Pseudomonas sp. YY3, respectively. The nitrification and denitrification enzyme genes could be detected using polymerase chain reaction (PCR). Moreover, the strain YY2 was a novel aerobic denitrifier belongs to genus of Sphingomonas, which showed great ability for metabolizing aromatic hydrocarbons. In the nitrification and denitrification process, the total nitrogen (TN) removal efficiency after 48 h was 94.22% and 90.10%, respectively. In the process of simultaneous nitrification and denitrification in mixed N-source, ammonia nitrogen was preferentially utilized. Furthermore, the strain YY2 exhibited excellent extracellular polymer secretion properties and excellent aerobic denitrification capacity using petroleum refractory organic compounds, which are beneficial for the formation of bacterial micelles and the engineering applications for the treatment of petrochemical wastewater.