Active packaging films have been an essential component in food material research to ensure the safe and efficient preservation of food, fruit, and vegetables. The shelf life of fruits and vegetables may likely be extended by covering them with high-performance nanofiber (NF) films. The selection of materials for active packaging film has been a critical factor in preventing food materials from environmental contaminants (microbes) and extending the shelf life. This study aims to develop NF-based materials for cherry tomatoes to prevent fungal and bacterial damage. Bioactive NFs were produced through an electrospinning process using tannic acid (TA) within a polyvinylidene fluoride (PVDF) template. These NFs offer a sustainable alternative to synthetic packaging for food preservation. TA was incorporated into the PVDF matrix at varying concentrations (0.4 to 1.2%). Key parameters, including moisture content, thickness, opacity, water-contact angle, and thermal shrinkage, were assessed. The physicochemical results indicate that the TA NFs are suitable for further shelf-life performance evaluations. The antifungal and antibiofilm activity of the NFs was tested, showing that the TA in the PVDF matrix was more effective than other concentrations. Shelf-life tests demonstrated that cherry tomatoes covered with TA NFs showed no surface changes for up to 4 days. Importantly, the NFs were confirmed to be non-toxic to normal cells, as evidenced by tests on mouse 3T3-L1 fibroblast cells. In summary, we have developed bioactive NFs composed of TA in a PVDF matrix that enhance the shelf life of cherry tomatoes by preventing bacterial and fungal attacks on the fruit surfaces.