Plant seeds are the ultimate source of nutrient-dense food, and also pharmaceutically valuable phytochemicals and bioactive components. Thus, this study was designed to evaluate the phytochemical profile, antimicrobial and antioxidant competence of various Macrotyloma uniflorum extracts using a standard in-vitro approach. The results obtained from the qualitative phytochemical analysis, revealed that the aqueous extract of M. uniflorum seeds contained a significant amount of phytochemicals such as flavonoids, phenols, tannins, saponins, steroids, terpenoids, protein, and carbohydrates. Interestingly, this aqueous extract showed remarkable antimicrobial activity against bacterial (Klebsiella pneumoniae, Staphylococcus aureus, and Escherichia coli) and fungal pathogens (Verticillium sp. and Aspergillus flavus) at more volume (100 µL). Similarly, a dose dependent antioxidant activity was recorded against number of free radicals such as DPPH (93.14%), and phosphomolybdenum (95.42%) by 150.0 µg mL-1 concentration. Furthermore, the IC50 values of these antioxidant assays were found as 59.75 & 59.77 µg mL-1 correspondingly. Thus, obtained results concludes that regular intake of M. uniflorum seed-based foods can reduce the deposition of reactive oxygen species and enhance cell viability, resulting in a maximum lifespan. More research is needed to identify the bioactive constituents held to account for antimicrobial and antioxidant activities