The goal of this study was to evaluate the antibiofilm and antimicrobial activities of Bacoside A, a mixture formulation of phytochemicals from Bacopa monnieri against Staphylococcus aureus and Pseudomonas aeruginosa which are known to form biofilm as one of their virulence traits. The antimicrobial effects of Bacoside A were tested using the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) assays. Cell membrane disruption assay was performed to find its possible target site. MTT assay, crystal violet assay and microscopic studies were performed to assess the antibiofilm activity.Bacoside A showed its antimicrobial activity against both the test organisms in their planktonic and biofilm states. It could significantly remove ~88-93% bacterial biofilm developed on microtiter plate at sub-minimum inhibitory concentration of 200 µg mL -1 .Biochemical and microscopic studies suggested that the eradication of biofilm might be due to the loss of extracellular polymeric substances and change in cell membrane integrity of the selected bacterial strains treated with Bacoside A. These results indicate that Bacoside A might be considered as an antimicrobial having ability to disrupt biofilms, thus it could alone or in combination with other therapeutics be useful to treat biofilm related infections caused by opportunistic bacterial pathogens.