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This study is aimed to develop bacterial cellulose (BC)-based biocompatible, biodegradable, bioactive, and non-toxic food packaging material. The preparation of BC/Ag nanocomposite was achieved through the reduction of silver nitrate with sodium chloride. Scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy, and X-ray diffraction (XRD) analyses confirmed the purity of BC and the development of BC/Ag nanocomposite. SEM analysis showed the uniform distribution of Ag nanoparticles in the BC matrix, which further improved the water solubility to 4.6% and tensile strength to 25.7 MPa of BC/Ag nanocomposite. The developed BC/Ag nanocomposite did not show any toxicity towards NIH-3T3 fibroblasts. The BC/Ag nanocomposite showed antimicrobial activity against three bacterial strains (Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli) and two fungal strains (Candida albicans and Trichosporon sp.) by producing inhibition zones of 0.17 cm, 0.08 cm, 0.16 cm, 0.06 cm, and 0.08 cm, respectively after 24 h. The BC/Ag nanocomposite film-coated oranges and tomatoes demonstrated acceptable sensory features such as odor and color at different storage temperatures for up to 9 weeks. These findings demonstrate that the BC/Ag nanocomposite film could be used as biocompatible packing material for providing protection and extending the shelf-life of different foods.