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A B S T R A C TBackground and Objectives: Microbial activity is the main factor in spoiling food products, which not only changes their texture and taste but also causes economic damage and poisoning. The present study aimed to assess the effects of essential oil of Bane (Pistacia atlantica) on physico-mechanical and antimicrobial properties of gelatin-carboxymethyl cellulose film. Materials and Methods:The solutions (4% w/v) of gelatin and CMC (1% w/v) were prepared in deionized water. Then different levels of essential oil (0, 03%, 0.6%, and 0.8%) were added to the solutions. Then the films were prepared by casting. The methods of physico-mechanical and antimicrobial evaluation were done based on previous research. Results:The results showed significant reduction in water vapor permeability, the thickness of film, tensile strength and solubility by increasing essential oil, while the percentage increased (P<0.05). In addition, L * , a* and b * were altered due to the color nature of essential oil of Bane. Increasing essential oil of Bane significantly inhibited the growth of Escherichia. coli, Salmonella enterica, Staphylococcus aureus, and Clostridium sporogenes. High concentrations of bane produced holes in the culture medium, the treatment containing 0.3% essential oil of bane only resulted in inhibition of Salmonella enterica, whereas higher concentrations of essential oil inhibited bacteria growth entirely.Conclusions: Essential oil of Bane improved the mechanical properties and inhibitory effects in regards to growth of microorganisms.