A two-chamber membraneless microbial fuel cell (MFC) was constructed, in which denitrifying bacteria attached on the cathode performed as denitrification by using electrons and protons supplied by anaerobic bacteria on the anode oxidizing organic pollutant and stainless steel meshes (the pitch of holes was 180μm) was adopted as the electrode. During the stable operation of the MFCs, the highest open cell voltage was 126.6 mV, and the maximum removal rate of COD, NH4 +-N, and NO3 --N was 88.9%, 99.7%, and 98.2%, respectively. However, the content of NO 2--N in the effluent was always lower than 1.25mg/L. The total and maximal removal rate for COD, NH4 +-N and NO3 --N in separate anode chamber and cathode chamber was 67.0%, 76.9%, and 84.0%, respectively, which were lower than that in the MFC. It is indicated that the two-chamber membraneless MFC has the collaborative and efficient capacity for COD and nitric pollutant removal by coupling oxidation on the anode and disoxidation on the cathode.