Nowadays, biodegradable antimicrobial packaging materials draw substantial attention and are considered one of the preferred emerging technologies to be used in the food industry. In this study, we have reported developing a two-component, chitosan (CS) and polyvinyl alcohol (PVA) based films loaded with natural extract of Ocimum tenuiflorum via solvent casting technique. Experimental results showed that all the components were well dispersed, resulting in complete homogenous films. FTIR-ATR spectral analysis indicated strong interaction between the film matrix components, which also gets reflected by the films' physical properties. Transparent biodegradable films have resulted from the mixture and incorporation of the natural agent/extracts influenced light barrier, water resistance, moisture content (9.89–9.01%), and DPPH radical scavenging properties (antioxidant value ~41.1%) of the CS/PVA films. Therefore, this natural extract containing transparent CS/PVA-based films promises to be used in the food industry as packaging material to enhance food safety.