Microbial Salinity Cell (MSC) can simultaneously desalinate water and generate electricity from the biodegradation of organic compound in wastewater. Utilization of a three-chambers configuration system along with electrolyte recirculation, creates a desalination process which occurs when the salt ions from the anode and cathode chambers are accumulated into the middle chamber, driven by the electrical energy generated from the organic compound biodegradation. The performance of three-chambers electrolyte recirculation MSC was investigated using three different NaCl concentrations of 2.0 g/L, 4.0 g/L, and 8.0 g/L, with the acetate concentration of 0.82 g/L. At 2.0 g/L NaCl, the maximum power density production was 42.76 mW/m 2 , increasing conductivity in the middle chamber from 15.09 µS/cm to 0.74 mS/cm. At 4.0 g/L, the maximum power density reached was 53.37 mW/m 2 , and conductivity in the middle chamber was raised from 60.08 µS/cm to 2.74 mS/cm. At 8.0 g/L, the power density was 29.29 mW/m 2 and conductivity in the middle chamber increased from 10.0 µS/cm to1.65 mS/cm. The performance of MSC was correlated with the initial NaCl concentration, with optimum NaCl concentration which was at 4.0 g/L, able to generate the highest power of 53.37 mW/m 2 and showed the highest increasing conductivity from 80.8 to 2.74 mS/cm.