In recent years, global energy systems have undergone a transition, with their energy matrices gradually shifting towards more renewable and cleaner sources. Among the various alternative energy sources, biofuels have become the subject of extensive academic research and scientific studies. The aim of this paper is to utilize jackfruit seeds, an agro-industrial residue, to obtain biofuel using the transesterification method. The oil was extracted from the seeds using the Soxhlet Extraction method, with hexane as the solvent. The jackfruit seeds exhibited an extractive content of 42.21 wt.%, which represents a significant portion of the biomass composition. Biodiesel production was achieved through transesterification with basic catalysis in ethanol, resulting in a conversion rate of 80.4%. The biofuel obtained was characterized by evaluating its physical-chemical parameters, including pH, electrical conductivity, acidity index, and density. The obtained results were satisfactory, and they met the specifications set by the ANP (Brazilian National Petroleum Agency) for biodiesel. Furthermore, they align with specifications found in the literature for other types of seeds. Therefore, it can be concluded that jackfruit seeds are a promising source for biofuel production. Additionally, the findings of this study provide motivation for further research in this field, particularly when synthesizing other types of seeds for biofuel extraction and comparing the results with those obtained from jackfruit seeds.