Background of the study: With rapid population growth, the demand and depletion of natural resources including fuel has surged leading to the exploration of renewable energy sources. Biofuels is one of the renewable energy sources and have the potential to act as alternative for fossil-based energy sources. Enormous efforts have been taken to enhance the production of biofuel and various methodologies are being continuously searched. Nanotechnology is one of the feasible techniques which contributes more in the biofuel industry. High cost and toxicity problems faced by physical and chemical synthesis methods of nanoparticles; route of green synthesis has been received considerable attention in world-wide. In general, green synthesis is highly effective, rapid, cheap and non-toxic and eco-friendly in nature. This method involves synthesis of nanoparticles using various biological organisms including microbes, lower and higher plant extracts and animal tissues and biomass. Aim: This review examines a comprehensive overview of the green nanotechnology to enhance biofuel production and address its associated challenges. Methodology: Data on green nanomaterials for biofuel production was collected until the year 2024 through the examination of research articles, patents and diverse online bibliographic databases such as Pubchem, Research Gate, PubMed, Science Direct, Scopus, Web of Science and Wiley Online Library. The online search was conducted using Green synthesized nanoparticles, biofuel application, production and challenges, upstream and downstream processes, biocatalysts and its environmental sustainability. Conclusion: This review is focused to collect information on the application of biofuel-production with its challenges and eco-friendly roles with green synthesised nanoparticles in bridging the gap between rising energy demand while maintaining the environmental sustainability.