This study presents the production of antibacterial Polyvinylpyrrolidone/cinnamon essential oil (PVP/CEO) nanofibers by oil-in-water emulsion electrospinning, with solution properties such as GC-MS, conductivity, viscosity, and characterization using SEM (fiber morphology), FT-IR (chemical structure), UV-vis (release properties of CEO). In addition, the antimicrobial properties of the nanofibers are also investigated by the disc diffusion method (antimicrobial properties) using Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Candida albicans. According to the results, the viscosity values increase with the addition of surfactant and with increased CEO concentration, but the conductivity increases until PVP1 then has decrease tendency. As concerns the fiber properties, the average fiber diameter increased with surfactant addition and CEO concentration except for PVP4. Also, the spinnability and fiber smoothness were enhanced significantly with surfactant and CEO. FT-IR analysis results verified the existence of CEO and PVP in the structure of the nanofibers. The absorbance values increased with the CEO concentration except for PVP4. Lastly, the antimicrobial results showed that 2, 3, and 4 wt% CEO concentration values are suitable for good antibacterial properties, while there is no zone formation with 1 wt% CEO concentration of PVP nanofibers. It is another important result that the antibacterial activity increases with the size of the nanofibrous material.