Microbial fuel cell (MFC) is an emerging biotechnology to convert the organic substrates in wastewater to electricity by anaerobic electrogenic bacteria. The main challenge for MFC research is to elucidate the fundamental mechanisms of electron generation and transfer and to apply these mechanisms to improve the power production in the engineering operation. This study extensively investigated the effects of three inocula (Geobacter sulfurreducens, soil, and wastewater) on the power production and electrochemical characteristics (i.e., internal resistances, Coulombic Efficiency) of MFCs. The results showed that the extents of bacterial adhesion varied between mixed cultures (soil) and pure cultures (G. sulfurreducens). The voltage output increased 30% when bacterial adhesion was well-developed in the soil inocula. Meanwhile, the inoculum types clearly affected the internal resistance (R-in) and power production of MFCs. Pure culture inoculum (G. sulfurreducens) had the lowest R-in (165 Omega) and the highest Coulombic Efficiency (CE, 25.8%) and Energy Conversion Efficiency (ECE, 7.2%), while the mixed culture inocula (soil) with the high concentration of nonelectrogenic bacteria, exhibited the highest R-in (620 Omega), lowest CE (9.2%) and lowest ECE (2.4%). Additionally, a second-order correlation was established between the anode potential (P-A) and power output while an exponential correlation was established between the difference between anode and cathode potentials (Delta PC-A) and power output.