•Electricity production from distillery wastewater was investigated in the MFC.•Mixed bacterial culture isolated from distillery wastewater and used as biocatalyst.•Cyclic voltammogram confirmed the exoelectrogenic activity of isolated culture.•The power density increased with increase in the anolyte pH from 6 to 8.•Performance of MFC increased with increasing COD concentration and conductivity. The effect of various system parameters such as wastewater Chemical Oxygen Demand (COD) concentration, pH, conductivity, membrane size and thickness on efficient energy production using mixed isolated culture from the distillery wastewater in the MFC was studied. The power density increased with increase in the anolyte pH from 6 to 8. The peak power density and COD removal efficiency was observed as 63.8±0.65mW/m2 and 63.5±1.5% at pH 8, respectively. The MFC performance increased with increasing COD concentration (800–3200mg/l), conductivity (1.1–9.7mS/cm) and membrane area (8–24cm2). The MFC operating with wastewater COD concentration of 3200mg/l and its conductivity of 9.7mS/cm produced the highest power density of 202±6mW/m2 with a corresponding current density of 412±12mA/m2. The results showed that the efficient electricity generation and simultaneous treatment of distillery wastewater can be attained in the MFC.