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A novel one step method was developed to dope nitrogen (N), sulfur (S) and carbon (C) in the Fe nanoparticles-dispersed carbon nanofibers (CNFs) grown over carbide-derived carbon (CDC), using thiourea as the single source of N, S and C. The synthesized N/S-Fe-CNF/CDC electrode was successfully used in a microbial fuel cell (MFC). When tested as the oxygen reduction reaction (ORR) catalyst, the electrode achieved a high current density (2.261 ± 0.002 mA/cm2), high OCP (0.611 ± 0.005 V), high stability upto 400 cycles, response time of ∼11 s, electron transfer number in the range 3.73–4.03, and Tafel slopes of −0.0627 and −0.183 V/dec at low and high current densities, respectively. A first order kinetics and a 4e− pathway were deduced from the ORR analysis. Notably, the fabricated MFC based on the prepared electrode produced a high current density of 1.3887 ± 0.002 mA/cm2, high OCP of 0.626 ± 0.005 V and maximum power density of 0.238 ± 0.002 mW/cm2, attributed to the synergistic effects of heteroatoms, Fe nanoparticles, and CNFs. Image 1 • Thiourea was used as the single source of heteroatoms (N, S) and carbon in Fe-CDC. • N/S-Fe-CNF/CDC-based MFC generated high OCP and power density. • Synergy of N, S, Fe, and graphitic CNFs resulted in the high performance of MFC. [ABSTRACT FROM AUTHOR]