Polymeric nanofiber readies an appropriate circumstance for enzyme immobilization for assortment of utilizations. In this examination, we’ve got made up polyvinyl alcohol /polyacrylonitrile (PVA/PAN) nanofibers utilizing electrospinning. After fabrication of nanofibers, the impact of atmospheric pressure air dielectric barrier discharge plasma (DBD) on PVA/PAN nanofibers were investigated in various times. Among them, 6min treat by air DBD plasma had the most significant impact on glucose oxidase (GOx) immobilization. Attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectrographic analysis and hydrophilic capacity and contact angle results uncovered that if there should arise an occurrence of DBD plasma modified nanofibers, the carboxyl bounds on a superficial level are expanded. The scanning electron microscopy (SEM) pictures indicated that, after GOx immobilization, the modified nanofibers with plasma has held its nanofiber structure. At last, we examined reusability and storage stability of GOx immobilized on plasma modified and unmodified nanofibers. The results were additional satisfactory for modified nanofibers with relevance to unmodified ones.