Microbial fuel cells (MFCs) are bio-electrochemical devices used to generate bioelectricity from a wide range of substrates by using bioelectrogenic microorganisms, and have special interest to supply the energy demands for small devices and the same time as a renewable energy source. In this study, a blend of soil-organic matter were tested in a membrane-less single chamber microbial fuel cells (SMFC) devices to generate bioelectricity along with biotransformation of organic wastes, over a testing time of 60 days and without renovation of substrate in batch mode of operation. The results in the experiments have shown relationship among substrate and the performance of output voltage in the SMFCs, the highest value of output voltage was 330 mV in the SMFC3 which contained a blend of 75:25 of fruits and vegetables.