The potential of deoiled Azolla pinnata biomass (DAB) as electrode and substrate was evaluated for microbial fuel cell (MFC) operation. The anode electrode was fabricated using biochar obtained by subjecting DAB to pyrolysis at 600 °C, while the reducing sugars after hydrolysis of DAB by acid pretreatment was used as substrate. The post pyrolyzed biochar (P-DAB) was characterized for structural and elemental functionalities using SEM, XRD and Raman spectroscopy, whereas the reducing sugar obtained from hydrolyzed DAB (H-DAB) was analyzed for its composition. Experimental results indicated that at a given 3 g COD/L resulted in a voltage of 382 mV with 65.6% of COD reduction in closed circuit (CC) mode of operation. Cyclic voltammetric analysis depicted maximum oxidative and reductive peak currents of 3.42 mA and -4.0 mA. Noticeable peaks were also identified in CC (-0.2 V to +0.2 V and -0.19 V to -0.3 V) and OC (+0.2 V to +0.4 V and -0.1 V to -0.3 V) corresponding to complex IV cytochrome c couples (cytochrome Cox (Cyt Cox)/cytochrome Crd (Cyt Crd)), signifying the participation of electron carriers during electron transfer. The microbiome diversity showed dominance of Proteobacteria, a phylum known for exo-electrogenic bacterial species. The DAB-derived products account to environmental sustainability and support circular bioeconomy in a biorefinery mode.