In this study, DOW CORNING 1-2577 Conformal Coating was proposed for the cathode diffusion layer of the microbial fuel cell (MFC). In MFCs, stainless steel mesh cathodes using DOW CORNING 1-2577 Conformal Coating/carbon as the diffusion layer and two poly (dimethylsiloxane) (PDMS)/carbon diffusion layers and carbon cloth cathode with four poly (tetrafluoroethylene) (PTFE) diffusion layers were constructed for comparison. Under the same operational condition, the MFCs with the DOW CORNING 1-2577 Conformal Coating/carbon diffusion layer produced the maximum power density of 1585±52?mW·m?2, compared with those using poly (tetrafluoroethylene) (PTFE) diffusion layers (1421±45?mW·m?2) and poly (dimethylsiloxane) (PDMS)/carbon diffusion layers (1353±49?mW·m?2). The DOW CORNING 1-2577 Conformal Coating could be an alternative for the diffusion layer construction in the MFC due to its remarkable performance and much simple construction procedure.