Herein, it was aimed to achieve antibacterial cellulose acetate (CA) nanofiber production and characterization. Firstly, solution properties, such as viscosity, conductivity and surface tension, were determined. Secondly, CA/zinc oxide (ZnO) composite nanofibers were produced with optimum process parameters via the electrospinning method. Then, the electrospun nanofibers were characterized by SEM, EDX, DSC, TGA, XRD, air permeability and water vapor permeability testing. Lastly, antibacterial activity tests were carried out in accordance with the AATCC100 method, against Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922).