In　this　paper,　the　influence　of　immobilized　bacteria　on　the　denitrification　performance　of　microbial　fuel　cells　(MFC)　was　studied,　and　a　novel　MFC　with　immobilized　bacteria　on　electrodes　was　developed.　The　anode　of　the　MFC　was　immobilized　with　electroactive　microorganisms,　while　the　cathode　was　immobilized　with　denitrifying　bacteria,　utilizing　water-based　polyurethane　as　a　carrier.　The　results　showed　that　the　electroactive　microorganisms　immobilized　on　the　anode　improved　the　electron　utilization　efficiency,　while　the　denitrifying　bacteria　immobilized　on　the　cathode　improved　the　denitrification　performance.　In　addition,　to　ensure　the　efficiency　of　MFC,　the　number　of　electrodes　can　be　reasonably　determined　according　to　the　number　of　immobilized　bacteria.　High-throughput　sequencing　analysis　showed　that　the　immobilization　of　bacteria　on　MFC　electrodes　was　beneficial　to　maintain　the　stability　of　the　functional　bacterial　community　structure.　Therefore,　the　biochemical　approach　of　utilizing　immobilized　bacteria　to　remove　nitrate　in　MFC　is　a　promising　technology.