The most prominent gram negative bacterium Pseudomonas aeruginosa that exist as a prevalent human pathogen emerged as a multidrug pathogen by expressing the formation of biofilm. These multiple drug resistant has become a major threat for current medical treatments. As the P.aeruginosa continues to develop resistance to the antibiotics due to its persistence in biofilm formation, the present research was designed to evaluate the anti-biofilm agents from mangifera indica and also its potentials as novel anti-pathogenic drug that targets Poly-beta- 1, 6-N- acetyl-D- glucosamine, an intercellular adhesin protein encoded by cupA gene, that plays a major role in the manifestation of genes involved in biofilm formation. In line with this, the 3D structure of Poly-beta- 1, 6-N- acetyl-D- glucosamine was modeled, validated and the potential binding sites were determined. The Phytochemical compounds of Mangifera indica were screened for their inhibitory activity by docking in the potential binding site of modeled Poly-beta- 1, 6-N- acetyl-D- glucosamine. The result of virtual screening revealed that all the compounds screened exhibited encouraging docking scores. The highest docking score was observed for Methylscopolamine (docking score – 20.3146 kJ/mol), which suggests that this compound might effectively occupy the binding regions; the results of these binding interactions indicate that the most important amino acids in the binding site of Poly-beta- 1, 6-N- acetyl-D- glucosamine are to be considered while designing the novel biofilm inhibitors that can effectively inhibit the MDR strains of Pseudomonas aeruginosa.