Microbial Fuel Cell (MFC) is a technology that converts chemical energy into electrical energy with catalytic reaction from microorganism. In this study, sugar mill effluent (SME) was used as the anodic substrate in a double chambered MFC for an application of electricity generation. Salt-bridge is the economic alternative to highly priced proton-exchange membrane in the construction of a microbial fuel cell. By altering the concentration of agar in the fabrication of salt-bridge, performance of microbial fuel cells was observed using sugar industrial wastewater as the substrate. In this study various concentrations of agar were taken in a salt bridge like 6gms, 8gms and 10gms and NaOH was also used in salt bridge for the flow of ions. Two different electrode materials like stainless steel rods and copper rods are used to generate electricity. Physiochemical analysis of parameters like TSS, TDS, Alkalinity, Acidity, BOD and COD has been carried out. It was observed that 8 gms of agar concentration with stainless steel rods has been reducing more pollutants when compared to 6gms and 10gms of agar with stainless steel and copper rods and maximum COD removal obtained was 56%.These results demonstrated that Sugar industrial effluent is a suitable substrate in a MFC for bioelectricity production and its treatment. Power output by the salt bridge MFC was 2.2mW/m2. The low power output was directly attributed to the higher internal resistance of the salt bridge system.