BACKGROUND: There has recently been increased interest in biodegradable and sustainable packaging within the food industry. Biopolymer materials based on renewable biomass can be used as alternatives to conventional plastic packaging. A corn protein, zein, possesses excellent film-forming properties because of its hydrophobic nature. It can be used for making edible films and for producing nanofibrous layers. Combination with polysaccharides like chitosan offers promising prospects for the production of delivery systems for the controlled release of active substances. The current trend is to minimize the content of chemical additives; thus essential oils are suitable alternatives to synthetic antimicrobials. RESULTS: This study aimed to develop various zein/chitosan-based film-forming solutions, films, and coatings with antimicrobial substances to prepare active food packaging. Thymol and three essential oils (thyme, cinnamon, oregano) were applied as bioactive ingredients against bacteria, yeasts, and fungi. The incorporation of these natural active compounds led to a decrease in particle size in most film-forming solutions and a reduction of zeta potential compared to controls. Release of the bioactive compound into an aqueous environment was proved by antimicrobial test. A zein/chitosan-based coating with thymol was applied on fresh strawberries. Microbiological analysis over 10 days confirmed the efficient control of bacterial and fungal growth. CONCLUSION: Zein/chitosan (7:1) systems are suitable as bioactive compound carriers to make barriers and to prevent moisture loss, ensuring microbial food quality and prolonging the shelf life of fruits. These systems can serve as sustainable active food packaging.