In this study, a membrane less microbial fuel cell with internal resistance of 4.9 MΩ was used in continuous mode for simultaneous generation of electricity and removal of organics from synthetic wastewater having equimolar concentration of glucose and glutamic acid. Activated sludge of a municipal wastewater treatment plant (MW‐AS) was used as the source of mixed microbial strains and graphite sheets were used as electrodes. The effects of process variables such as empty bed contact time, electrode surface area, inter electrode distance, mediator concentration etc., on the voltage generation and color and COD removal have been studied. Under optimum conditions with initial COD value of 500 mg/L, the maximum voltage and current have been found to be ∼ 200 mV and ∼ 2.5 mA, respectively, with the COD removal of 90%. © 2015 American Institute of Chemical Engineers Environ Prog, 34: 1404–1413, 2015