Electrochemically active anodic biofilm that has adapted under mild applied potentials in the range 100-500 mV was evaluated for its improved bioelectrogenesis and bioelectrochemical treatment of petroleum refinery wastewater (PRW) in a single chamber air cathode microbial fuel cell (MFC). MFC operation with 500 mV as supplemental voltage has exhibited a maximum power density of 132 mW/m2, which was three times higher than control MFC (45 mW/m2). Similarly, highest substrate removal efficiency (48%) was also obtained with the MFC of 500 mV, followed by 300 mV (37%), 100 mV (32%) and control (27%). Adaptation under applied potential conditions also exhibited enhanced degradation efficiency of diesel range organics (DROs)/straight chain-alkanes. The strategy efficiently reduced DROs with the maximum efficiency of 89% (500 mV), which is almost 50% higher than that of the control system (59%), demonstrating the effectiveness of using supplemented voltage in treating PRW.