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The development of new materials environmentally friendly has become an important market niche for the food industry. The agro-industrial wastes and by-products can be an alternative for the production of biodegradable food packaging. The work aimed to produce biodegradable plastic film extruded with antioxidant and antibacterial properties by the joining of cassava starch and Brazilian propolis by-product (BPB). The volatile profile of BPB, Brazilian propolis by-product film (BPBF), and control film (CF) were analyzed by gas chromatography. The mechanical, antioxidant, and antibacterial properties of the films were also assessed. Eighty-seven volatile compounds were detected with aldehydes and terpenoids predominating in the samples. The major terpenoid detected in the samples was the 2-pinen-10-ol followed by α-copaene. Benzaldehyde and benzenepropanoic acid ethyl ester were the major aldehyde and ester classes present in the films. The BPBF exhibited promising mechanical properties showing the highest Elasticity modulus (11.96 Mpa) and excellent antioxidant (8.45 mmol Trolox equivalent/g) and antibacterial (12.5 mg/mL against Staphylococcus aureus and Salmonella Typhimurium) activities. The valorization of agro-industrial by-products following the current trends to environmental care can be a sustainable alternative for the development of a plastic into which the propolis by-product is incorporated in biodegradable packaging info:eu-repo/semantics/publishedVersion