Silver nanoparticles (AgNPs) are effective antimicrobial agents however excess release of silver causes argyria and argyrosis. An strategy to avoid these detrimental side effects is immobilization of AgNPs on several organic and inorganic substrates. Herein, we propose immobilization of AgNPs on TiO nanoparticles by an environmentally green process subsequently incorporating the TiO/AgNP into cellulose acetate (CA) nanofiber matrix. The TiO/AgNP nanocomposite particles were prepared by coating TiO nanoparticles with polydopamine hydrochloride followed by a treatment in AgNO solution. Subsequently, the TiO/AgNP nanocomposites were added into CA solution and electrospun to fabricate CA/TiO/AgNP composite nanofibers. The samples were characterized by XRD, TEM, XPS, SEM, EDX, FTIR and antibacterial assays. Synthesis of TiO/AgNP and its loading into CA nanofibers was confirmed by XRD, XPS, TEM and EDX analysis. SEM images indicated regular morphology of the nanofibers. The antibacterial test results confirmed CA/TiO/AgNP composite nanofibers having excellent antibacterial performances for 36 h and substantial bacterial growth inhibition for 72 h.