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Introduction This study investigates the health-promoting properties of fatty acids isolated from Vicia faba (broad beans) seeds, focusing on their potential as natural bioactive compounds for managing oxidative stress, microbial infections, and metabolic disorders. Given the global rise in chronic diseases such as diabetes and cardiovascular conditions, there is a pressing need for alternative treatments derived from plant sources. Fatty acids from Vicia faba seeds were extracted and analyzed for their antioxidant, antimicrobial, and anti-diabetic activities using in vitro assays.Materials and Methods Vicia faba seeds were dried, ground into powder, and subjected to Soxhlet extraction to isolate fatty acids. The isolated fatty acids were analyzed using gas chromatography-mass spectrometry (GC-MS) to identify their composition. Antioxidant activity was measured using the DPPH radical scavenging assay, while antimicrobial activity was assessed against bacterial strains such as Staphylococcus aureus and Escherichia coli through the disc diffusion method. Enzyme inhibitory assays were conducted to evaluate the inhibition of α-amylase and α-glucosidase, which are associated with postprandial glucose control.Results The results revealed that the isolated fatty acids possess strong antioxidant properties, with an observed DPPH radical scavenging activity of 78% at a concentration of 200 µg/mL, indicating high efficacy in reducing oxidative stress. Additionally, antimicrobial testing showed that the fatty acid extracts exhibited significant inhibition zones against Escherichia coli and Staphylococcus aureus, with inhibition diameters of 18 mm and 20 mm, respectively, at 100 µg/mL concentration. In enzyme inhibition assays, a notable reduction in α-glucosidase activity was recorded, achieving 65% inhibition at 150 µg/mL, underscoring the potential of these fatty acids in controlling postprandial blood glucose levels.Conclusion The findings suggest that fatty acids derived from Vicia faba seeds have promising health benefits that can be harnessed for natural therapeutic applications, especially in the management of metabolic disorders and infections. This research contributes valuable insights into the bioactivity of Vicia faba fatty acids, supporting their use in developing natural remedies for chronic disease management.