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In this work, Poly(lactic acid) (PLA)/poly(butylene carbonate) (PBC)/graphene oxide (GO) nanofiber membranes with different content GO were successfully prepared by electrospinning. The structural and physical characteristics of the membranes were investigated by scanning electron microscopy (SEM), Differential Scanning Calorimeter (DSC), contact angle test, Fourier Transform Infrared Spectroscopy (FT-IR), agar diffusion method. SEM observations revealed a fine dispersion of GO in the nanofiber matrices. FT-IR spectra result showed that the GO, PBC and PLA mixed well in the PLA/PBC/GO films. The antibacterial properties of nanofiber membranes were also investigated. The results indicated that the antimicrobial activities of PLA/PBC/GO composite nanofibrous films were much higher than that of pure PLA/PBC nanofibrous films. Meanwhile, the antimicrobial activity against S.aureus was also slightly higher than E. coli.