Generally, high bioelectroactivity of anodophilic biofilm favors high power generation of microbial fuel cell (MFC); however, it is not clear whether it can promote denitrification of MFC synchronously. In this study, we studied the impact of anodophilic biofilm bioelectroactivity on the denitrification behavior of air‐cathode MFC (AC‐MFC) in steady state and found that high bioelectroactivity of anodophilic biofilm not only favored high power generation of the AC‐MFC, but also promoted the growth of denitrifers at the anodes and strengthened denitrification. Anodophilic biofilms of AC‐MFC with various bioelectroactivity were acclimated at conditions of open circuit (OC), Rext of 1000 Ω and 20 Ω (denoted as AC‐MFC‐OC, AC‐MFC‐1000Ω, and AC‐MFC‐20Ω, respectively) and performed for over 100 days. Electrochemical tests and microbial analysis results showed that the anode of the AC‐MFC‐20Ω delivered higher current response of both oxidation and denitrification and had higher abundance of electroactive bacteria than the AC‐MFC‐OC, AC‐MFC‐1000Ω, demonstrating a higher bioelectroactivity of the anodophilic biofilms. Moreover, these electroactive bacteria favored the accumulation of denitrifers, like Thauera and Alicycliphilus, probably by consuming trace oxygen through catalyzing oxygen reduction. The AC‐MFC‐20Ω not only delivered a 61.7% higher power than the AC‐MFC‐1000Ω, but also achieved a stable and high denitrification rate constant (kDN) of 1.9 h−1, which was 50% and 40% higher than that of the AC‐MFC‐OC and AC‐MFC‐1000Ω, respectively. It could be concluded that the high bioelectroactivity of the anodophilic biofilms not only favored high power generation of the AC‐MFC, but also promoted the enrichment of denitrifers at the anodes and strengthened denitrification. This study provided an effective method for enhancing power generation and denitrification performance of the AC‐MFC synchronously.