Microbial fuel cells (MFCs) represent simple devices that harness the metabolic activities of microorganisms to produce electrical energy from diverse sources such as organic waste and sustainable biomass. Because of their unique advantage to generate sustainable energy, through the employment of biodegradable and repurposed waste materials, the development of MFCs has garnered considerable interest. Critical elements are typically the electrodes and separator. This mini-review article presents a critical assessment of nanofiber technology used as electrodes and separators in MFCs to enhance energy generation. In particular, the review highlights the application of nanofiber webs in each part of MFCs including anodes, cathodes, and membranes and their influence on energy generation. The role of nanofiber technology in this regard is then analysed in detail, focusing on improved electron transfer rate, enhanced biofilm formation, and enhanced durability and stability. In addition, the challenges and opportunities associated with integrating nanofibers into MFCs are discussed, along with suggestions for future research in this field. Significant developments in MFCs over the past decade have led to a several-fold increase in achievable power density, yet further improvements in performance and the exploration of cost-effective materials remain promising areas for further advancement. This review demonstrates the great promise of nanofiber-based electrodes and separators in future applications of MFCs.