To improve cathodic efficiency and sustainability of microbial fuel cell (MFC), graphite fibre brush (GFB) was examined as cathode material for power production in biocatalysed-cathode MFC. Following 133-h mixed culturing of electricity-producing bacteria, the MFC could generate a reproducible voltage of 0.4 V at external resistance (REX) of 100 Ω. Maximum volumetric power density of 68.4 W m–3 was obtained at a current density of 178.6 A m–3. Upon aerobic inoculation of electrochemically active bacteria, charge transfer resistance of the cathode was decreased from 188 to 17 Ω as indicated by electrochemical impedance spectroscopy (EIS) analysis. Comparing investigations of different cathode materials demonstrated that biocatalysed GFB had better performance in terms of half-cell polarisation, power and Coulombic efficiency (CE) over other tested materials. Additionally, pH deviation of electrolyte in anode and cathode was also observed. This study provides a demonstration of GFB used as biocathode material in MFC for more efficient and sustainable electricity recovery from organic substances.