<p>This study investigates the effects of 3D nitrogen-doped graphite anodes (NG) in different orientations within the anode chamber in the microbial fuel cell (MFC). The performance of MFC was evaluated based on power density, COD removal efficiency measurements, and the impact of different anode configurations on system stability. X-ray diffraction (XRD) exhibited the changes in crystallite size due to the incorporation of nitrogen into the graphite structure. Raman spectroscopy revealed the increase in defects because of the effective integration of nitrogen. Energy-dispersive X-ray spectroscopy (EDX) confirmed the existence of nitrogen content and a successful nitrogen doping procedure. Additionally, scanning electron microscopy (SEM) revealed an enhancement in the layer-like structure of graphite and introduced defects to the surface area of the NG anodes. Also, electrochemical analyses exhibited that the NG anodes outperformed untreated graphite anodes (G) due to increased specific surface area and enhanced electron transfer efficiency. MFCs with VNG and HNG anodes achieved the highest power density output, reaching 607 ± 25 mW/m² and 537 ± 14 mW/m², respectively. These values are significantly higher than the power density of MFC with G anodes, which was 318 ± 12 mW/m². The lowest total internal resistance was observed in MFCs with HNG and VNG anodes, measuring 92 ± 10 Ω and 127 ± 7 Ω, respectively, which were 87 to 52 Ω lower than the internal resistance of the MFC with G (179 ± 25 Ω). Similarly, the charge transfer resistance (<i>R</i>_ct) was specified by electrochemical impedance spectroscopy (EIS) in MFCs. The <i>R</i>_ct values for the VNG and HNG anodes were 41.26 Ω.cm² and 50.48 Ω.cm², respectively, representing a reduction of 47% and 35% compared to the G anode (78.76 Ω.cm²). The results show that the NG anodes significantly increase the power density and wastewater treatment efficiency. Furthermore, the MFCs with nitrogen-doped graphite anodes exhibited enhanced chemical oxygen demand (COD) removal efficiency due to improved biofilm activity. The COD removal efficiencies of VNG and HNG anodes achieved 70.14% ± 5.23% and 65.83% ± 3.89%, respectively, compared to 62% ± 4.49% for the G anode. According to the outcome, this method is introduced as a reliable solution for energy and water sustainable management. This research helps optimize the MFC systems for real-world applications in wastewater treatment and renewable energy generation and also aligns with global sustainable development.</p> Graphical abstract <p></p>