To determine the antioxidant and antibacterial abilities of the polyphenol rich fraction form leaves of Ocimum gratissimum. The total polyphenol content was evaluated using Folin-Ciocalteu assay. ABTS, lipid peroxidation, superoxide and nitric oxide scavenging capacity assays were performed to test the antioxidant activity. Subsequently, the antimicrobial activities against Bacillus subtilis, Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa bacterial strains were assessed using disc diffusion methods. Results of polyphenol rich fraction form leaves of Ocimum gratissimum contained high total polyphenol content. The polyphenol rich fraction form leaves of Ocimum gratissimum showed effective antioxidant capacity EC50 value of ABTS, lipid peroxidation 54.23 and 65.45µg/mL respectively. Additionally, the flavonoid rich fraction was effective against both Gram-positive bacteria. In conclusion flavonoid rich fraction from leaves of O. gratissimum demonstrated valuable antioxidant and antimicrobial properties.