Wound healing is a complex process in which bacterial infection is a major cause for delayed wound healing. Occurrence of drug resistance among bacterial pathogens led to discovery of new antimicrobial agents from new sources. The present study aimed to identify microalgal metabolites with antibacterial activity and to develop a wound dressing film with their potential healing activity. Microalgal samples were collected from three different freshwater habitats, isolated, made them pure cultures and physico-chemical properties of water samples from the respective sampling sites were analzed. Among the three microalgal isolates, Chlorella sp. NRMC-F-0350 showed antibacterial activity against clinical isolates of E. coli, Klebsiella pneumoniae and Staphylococcus aureus. Additionally, functional groups present in Chlorella sp. NRMC-F-0350 were identified using FTIR and GC-MS analysis. Several antibacterial compounds viz. benzoxazole, 1,2 Benzene dicarboxylic acid, sistosterol, 9-Octadecanoic acid, eicosane and hexadecane were identified. Wound dressing films were developed and showed evident antibacterial as well as significant wound healing activity (84.5%). Therefore, the developed films can be used as a potential wound dressing material.