Abstract Background Carbon-based materials are commonly used as anodes in microbial fuel cells (MFCs), whereas metal and metal-oxide-based materials are not used frequently because of low electrical output. Stainless steel is a low-cost material with high conductivity and physical strength. In this study, we investigated the power generation using flame-oxidized (FO) stainless steel anodes (SSAs) in single-chambered air-cathode MFCs. The FO-SSA performance was compared to the performance of untreated SSA and carbon cloth anode (CCA), a common carbonaceous electrode. The difference in the anodic community structures was analyzed using high-throughput sequencing of the V4 region in 16S rRNA gene. Results Flame oxidation of SSA produced raised node-like sites, predominantly consisting of hematite (Fe2O3), on the surface, as determined by X-ray diffraction spectroscopy. The flame oxidation enhanced the maximum power density (1063 mW/m2) in MFCs, which was 184 and 24 % higher than those for untreated SSA and CCA, respectively. The FO-SSA exhibited 8.75 and 2.71 times higher current production than SSA and CCA, respectively, under potentiostatic testing conditions. Bacteria from the genus Geobacter were detected at a remarkably higher frequency in the biofilm formed on the FO-SSA (8.8–9.2 %) than in the biofilms formed on the SSA and CCA (0.7–1.4 %). Bacterial species closely related to Geobacter metallireducens (>99 % identity in the gene sequence) were predominant (93–96 %) among the genus Geobacter in the FO-SSA biofilm, whereas bacteria with a 100 % identity to G. anodireducens were abundant (>55 %) in the SSA and CCA biofilms. Conclusions This is the first demonstration of power generation using an FO-SSA in MFCs. Flame oxidation of the SSA enhances electricity production in MFCs, which is higher than that with the common carbonaceous electrode, CCA. The FO-SSA is not only inexpensive but also can be prepared using a simple method. To our knowledge, this study reveals, for the first time, that the predominant Geobacter species in the biofilm depends on the anode material. The high performance of the FO-SSA could result from the particularly high population of bacteria closely related to G. metallireducens in the biofilm.