Electrospun nanofibrous membranes have gained great focused in medical research due to its simplicity, diversity and biodegradability. The challenge to researchers is to make more effective and sustainable by incorporating pristine materials to address adverse health issues of society. In this work, considering the unique characteristics of Graphene and its derivatives, well-dispersed Graphene Oxide (GO) were prepared using Modified Hummer's method. Further, the mixture of solutions, Poly (Vinyl Alcohol) PVA with synthesized GO nano-particles, was successively fabricated into nanofibrous membranes by electrospinning technique. Further, the electrospun membranes were cross-linked through vapours of Glutaraldehyde (GA) in controlled environment to make membranes hydrophobic in nature. In addition, the characterization of synthesized GO and electrospun nanofibers were done using SEM, XRD and FTIR. The results show that GO incorporation decreases the average diameter of nanofibers from 422±133nm to 274.1±93.23nm, whereas crosslinking of nanofibers at various hours (12 to 48h) tends to increase the average diameter from 368.4±130.1nm to 671.41±293nm. In addition to that the 12h crosslinked nanofibers membrane shows better antibacterial activity than without crosslinked PVA/GO membrane against E. coli after 24h of incubation. This primarily work provides a basis for further studies of this novel nanofibrous material.