Abstract: The rising resistance of bacteria to antibiotics has driven the search for new antimicrobial agents. This study focused on encapsulating Buforin I, an antimicrobial peptide, in chitosan/polyethylene oxide (CS-PEO) nanofibers. Buforin I was loaded at a minimum bactericidal concentration (MBC), 10× MBC, and 20× MBC, with assessments on morphology, thermal properties, chemical bonds, crystalline structure, mechanical strength, antimicrobial activity, and cell toxicity. Techniques like differential scanning calorimetry and Fourier-transform infrared spectroscopy confirmed the effective loading of Buforin I in the nanofibers. Scanning electron microscopy showed that Buforin incorporation increased nanofiber diameters. The tensile strength peaked at 20× MBC. Microbial tests indicated that the inhibition zone for nanofibers at 20× MBC surpassed that of commercial antibiotics. Beef coated with CS-PEO nanofibers containing Buforin I demonstrated reduced pH and water activity, alongside lower weight loss during storage. Texture and color analyses revealed that the Buforin I nanofibers helped maintain beef hardness and slowed color degradation compared to control samples. Moreover, thiobarbituric acid levels and total microbial counts in the coated beef were significantly lower than controls (below 3 log CFU/g after 9 days at 4 ◦C). Thus, these nanofibers may serve as effective antimicrobial packaging agents to delay food spoilage.

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