The present study reported, for the first time, the fabrication and characterization of electrospun nanofibers based on arabinoxylans (AXs) alone. The Fourier transform infrared spectrum of ferulated water-extractable AXs recovered from wheat endosperm confirmed the molecule identity. The carbon and oxygen signals in X-ray photoelectron spectrometry (XPS) were recorded for this molecule. The AXs had weight-average molar mass, intrinsic viscosity, radius of gyration, and hydrodynamic radius values of 769 kDa, 4.51 dL/g, 55 nm, and 31 nm, respectively. The calculated AX characteristic ratio and persistence length were 10.7 and 3.2 nm, respectively, while the Mark–Houwink–Sakurada α and K constants were 0.31 and 9.4, respectively. These macromolecular characteristics indicate a molecular random coil structure in the polysaccharide. Using aqueous acetic acid 50% (v/v) as a solvent favored the Taylor cone establishment and the fabrication of electrospun nanofibers. The morphology of nanofibers was revealed by scanning electron microscopy images. Atomic force microscopy analysis of AX nanofibers exposed the material deposition in layers; these nanofibers had an average diameter of 177 nm. These nanofibers could be used as advanced biomaterials for biomedical applications such as wound dressing.