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The present study explores the use of carbon dots coated with Iron (II, III) oxide (Fe₃O₄) for its application as an anode in microbial fuel cells (MFC). Fe₃O₄@PSA-C was synthesized using a hydrothermal-assisted probe sonication method. Nanoparticles were characterized with XRD, SEM, FTIR, and RAMAN Spectroscopy. Different concentrations of Fe₃O₄- carbon dots (0.25, 0.5, 0.75, and 1 mg/cm²) were coated onto the graphite sheets (Fe₃O₄@PSA-C), and their performance in MFC was evaluated. Cyclic voltammetry (CV) of Fe₃O₄@PSA-C (1 mg/cm²) modified anode indicated oxidation peaks at −0.26 mV and +0.16 mV, respectively, with peak currents of 7.7 mA and 8.1 mA. The fluxes of these anodes were much higher than those of other low-concentration Fe₃O₄@PSA-C modified anodes and the bare graphite sheet anode. The maximum power density (Pmax) was observed in MFC with a 1 mg/cm² concentration of Fe₃O₄@PSA-C was 440.01 mW/m², 1.54 times higher than MFCs using bare graphite sheet anode (285.01 mW/m²). The elevated interaction area of carbon dots permits pervasive Fe₃O₄ crystallization providing enhanced cell attachment capability of the anode, boosting the biocompatibility of Fe₃O₄@PSA-C. This significantly improved the performance of the MFC, making Fe₃O₄@PSA-C modified graphite sheets a good choice as an anode for its application in MFC.