In this study, natural silk web and natural silk non-woven fabric were prepared mechanically using the binding character of the sericin in silk. The effect of process variables on the preparation, structure, and properties of the silk web and the non-woven fabric was examined. The reeling velocity affected the morphology and mechanical properties of the web but had almost no influence on the crystalline structure of the silk. From the viewpoint of reel-ability and the mechanical properties (work of rupture) of silk web, a reeling velocity of 39.2 m/min represented the optimal processing velocity. The porosity and swelling ratio of the silk web decreased slightly with increasing reeling velocity. Furthermore, the reeling bath temperature had a significant effect on the reel-ability of silk filaments from a silkworm cocoon. Bath temperatures ≥50 °C yielded good reel-ability (>900 m reeling length). The porosity, swelling ratio in water, and mechanical properties of the silk web and silk non-woven fabric changed only slightly with the reeling bath temperature but changed significantly with the hot press treatment. The hot-pressed silk web (i.e., silk non-woven fabric) exhibited higher tensile strength as well as lower elongation at break, porosity, and swelling ratio than the silk web.