A versatile and effective method of producing polyvinyl alcohol (PVA)-xylanase (XY) fibers cross-linked by glutaraldehyde vapor (GA) is reported in the present study. Crosslinking was performed for 30, 60, 90, and 180 min. The morphology, structure, and thermal stability of the fibers were investigated by scanning electron microscopy (SEM), Fourier-transformed infrared spectroscopy (FTIR), and thermogravimetric analyses (TGA). The enzymatic activity was studied under different storage time and reutilization cycles. The enzyme's ability to reduce the turbidity of a standard beta-glucan solution was also verified. Cross-linked fibers presented denser and more compacted structures. There was an improvement in thermal properties as well as in the recycling and storage efficiency of cross-linked XY-PVA fibers, compared to their non-cross-linked counterparts. Free and 180 min-cross-linked immobilized enzymes were able to reduce the turbidity of the beta-glucan solution by a similar degree.