In this study we prepared a nanocomposite substrate from poly lactic acid (PLA) – Graphene oxide (GO) using electrospinning to study mechanical properties, surface chemical structure and topology. We employed different methods to characterize and prepare the substrate for enzyme immobilization. Surface topology and chemical structure were important for enzymes binding to the substrate surface. GO facilitated the electrospinning process, and effect of its presence on immobilized enzyme stability and activity was investigated here for the first time here. Scanning electron microscopy images of nanofibers confirmed the presence of fine fibers in membrane morphology. Use of PLA as a biocompatible substrate and effect of GO for enzyme stability modification produces an acceptable substrate for biotechnological applications such as growth factor enzyme immobilization Keywords: Nanocomposite, Electrospinning, PLA, GO, Enzyme Immobilization