Infections caused by the biofilm-forming multi-drug resistant organisms are increasing day by day. In this study, polyvinyl alcohol (PVA) based antimicrobial membranes were developed with the incorporation of lipopeptides (LPs) and zinc oxide nanoparticles (ZnONPs) by solvent casting method. The fabricated membranes were further characterised by the FT-IR, SEM, UV-vis spectroscopy, TGA and mechanical analysis. The results showed enhanced optical, thermal, mechanical and water barrier properties for the membranes engineered with LPs and ZnONPs. The membranes also showed excellent activity against the biofilm-forming bacteria such as the Staphylococcus aureus, Klebsiella pneumoniae and Pseudomonas aeruginosa. Also, the developed nanocomposite membranes were found to be hemocompatible with enhanced dressing pH features. Hence, the developed nanocomposite membranes can be considered to have promising biomedical application as antimicrobial coating.