In this study, an active packaging containing Zataria multiflora essential oil (ZMEO), a powerful natural antimicrobial agent, encapsulated into polyvinyl alcohol (PVA) fiber via electrospinning is presented. ZMEO was effective on pathogenic bacteria, particularly Gram-positive bacteria (Staphylococcus aureus, Bacillus cereus, Listeria monosytogene), fungi and yeasts (Aspergillus fumigatus, Candida albicans). Results showed that the scanning electron microscopy (SEM) images of fibers had a bead-free and uniform structure. Fourier-transform infrared (FTIR) revealed that ZMEO was encapsulated into PVA through a physical process, without chemical interaction between the ingredients. Strawberries treated with PVA/ZMEO significantly (p < 0.05) preserved the anthocyanin (18.64%), total phenols (12.95%), antioxidant (22.72%), soluble solids (6.44%), titratable acidity (20.88%), firmness (27.2%), and color (15.55%) compared to the control sample during 15 days of cold storage. According to these findings, electrospinning was an efficient method for encapsulating bioactive compounds. ZMEO loaded into PVA fiber delayed the physiological and biochemical changes of fruits and extended the fruit’s shelf-life. This study revealed the benefits of incorporating ZMEO into PVA fiber mats, which could lead to new possibilities for active packaging.