CuO nanoparticles (NPs) have been used for the antimicrobial agent against different pathogenic microorganisms. In this study, CuO NPs are immobilized on the surface of activated carbon fiber (ACF) with the enhancement of (3-aminopropyl)triethoxysilane (APTES) as an organic binder. The obtained fibers are evaluated by coating efficiency, structural deformation, and antimicrobial activities. In the results, APTES can improve the immobilization of CuO on the surface of ACF. Also, the curing of silane layers at high temperature leads to the high coating efficiencies as well as structural reinforcement. The samples with drying step after APTES coating step (denoted as DA-CuO) have the highest antimicrobial activity against both Escherichia coli and Staphylococcus aureus after 24 hours treatment, respectively.