Feasibility of a bio-electrochemical process for simultaneous nitrification and denitrification in a single reactor was experimentally investigated. The reactor consisted of anodic and cathodic electrodes, on which nitrifying and denitrifying biofilms, respectively, were fixed. Effects of the applied electric current and DO concentration in the bulk solution were examined. The TN removal could be achieved through the occurrence of nitrification and denitrification in anodic and cathodic biofilms, respectively. Both nitrification and denitrification rates increased with an increase in the applied electric current. Even at low DO concentration in the bulk solution, nitrification proceeded at a high rate by utilizing oxygen generated on the anode. Denitrification rate remained relatively high at high DO concentration due to supplying hydrogen gas to the inner side of the cathodic biofilm. The higher TN removal rate tended to be obtained at lower DO concentration and higher current density. From these results, it was concluded that the bio-electrochemical process was applicable to simultaneous nitrification and denitrification due to stable formation of aerobic and anoxic regions in the single reactor.