The present study included the environmentally friendly production of stable nickel nanoparticles (NiO NPs) using lemon and tomato, followed by their analysis and evaluation for their antibacterial properties against Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, and Bacillus cereus. The Nickel oxide nanoparticles produced exhibited their maximum absorption at 276 nm in the UV-vis spectrum. The image captured FESEM revealed smooth nanofibers with an average diameter of around 259 ± 3.7 nm. X-ray diffraction (XRD) experiments verified the existence of elemental nickel and accurately determined the crystalline structure of nickel oxide nanoparticles. Novel green organic-inorganic hybrid nanofibers (NiO@CA) were synthesized using the electrospinning technique. These nanofibers are composed of NiO nanoparticles integrated into cellulose acetate nanofibers, which are particularly engineered for the purpose of fruit preservation. Robust antibacterial activity was shown by NiO nanoparticles and NiO@CA nanofiber against the assessed food pathogenic bacterial strains. NiO@CA nanofiber used as a surface coating on lemon and tomato prolonged their shelf life by preventing the degradation caused by food risks. The provided results indicate that NiO@CA nanofiber has the capacity to function as antimicrobial packaging for the purpose of food preservation. Enhancing food safety, prolonging shelf life, and offering an eco-friendly alternative to traditional materials, antibacterial food packaging made of biocompatible nickel oxide-infused cellulose acetate electrospun nanofibers is a great solution.