The stimulatory effect of conductive particles on anaerobic digestion has been demonstrated in recent years. However, it is yet to be determined whether and how conductive particles affect methanogenesis via electro-fermentation (electro-fermentation methanogenesis). In this study, it was demonstrated, for the first time, that conductive graphite boosted the methane production yield by 54.3% and increased the maximum methane production rate by 72.2% during electro-fermentation methanogenesis. Graphite significantly affected the composition of cathode biofilms, with more live and large aggregates being observed. Spectroelectrochemical analyses further showed that the kinds and intensities of biocatalytic active sites and redox groups on the cathode biofilms increased during graphite-assisted electro-fermentation methanogenesis. Particularly, c-type cytochromes, humic acid-like substances, and humic substances improved the long-range electron transport to methanogens such as Methanobacterium and Methanosarcina. The results have implications for the improvement of electro-fermentation process and the use of conductive materials for biofuel recovery.