This work assessed the performance of a single-chamber microbial fuel cell (MFC) with various substrates. Primary settled domestic wastewaters were used to simulate wastewaters of high biodegradability; while phenol-based wastewaters and benzene-based wastewaters were used to simulate wastewaters of low biodegradability. Experiments were performed at initial pH values of 6, 7 and 8. The maximum voltage production, power density and removal of substrate were obtained using primary settled domestic wastewater, whereas the lowest values were obtained using phenol-based wastewater. The maximum chemical oxygen demand removal efficiency, phenol removal efficiency and benzene removal efficiency were 80.8, 63.3 and 77.8%, respectively. The performance of the MFC was enhanced by increasing the influent pH. The lowest coulombic efficiencies were obtained from phenol-based wastewater and benzene-based wastewater, which indicated that electrogenic bacteria were not the primary microorganisms responsible for the biodegradation of low biodegradable wastewater.