The development of antimicrobial agents derived from micro-organisms and chemotherapeutic agents from plants is a research area of the utmost importance. The present study was designed to evaluate the antimicrobial activity of banana (Musa sapientum) Blossom extract against Gram-positive and Gram-negative bacteria (Bacillus subtilis, Bacillus cereus and Escherichia coli) with extract-incorporated Chitosan (CS) - Polyethylene Glycol (PEG) blended film. The extraction was carried out in ethanol, chloroform and water. The antimicrobial activities of the extracts evaluated using disc methods and Minimum Inhibition Concentration (MIC).The compounds responsible for antimicrobial activity was conformed to confirmatory test. Ethanol extract showed an antibacterial activity against the tested microorganisms.