Hollow tubular conduits (TCs) with tunable architecture and biological properties are in great need for modulating cells functions and drug delivery in guided tissue regeneration. Herein, a new methodology to produce enzymatically-crosslinked silk fibroin TCs is described, which takes advantage of the tyrosine groups present in silk structure that are known to allow the formation of a covalently-crosslinked hydrogel. Three different processing methods were used as a final step to modulate the properties of the silk-based TCs. This approach allows to virtually adjust any characteristic of the final TCs. The final microstructure ranged from a non-porous to a highly porous network, allowing the TCs to be selectively porous to 4 KDa molecules, but not to human skin fibroblasts. Mechanical properties were dependent both on the processing method and thickness of the TCs. Bioactivity was observed after 30 days of immersion in Simulated Body Fluid only for the TCs submitted to a drying processing method (50ºC). The in vivo study performed in CD-1 Albino mice demonstrated the good biocompatibility of the TCs. The enzymatically-crosslinked silk fibroin TCs are versatile and have adjustable characteristics that can be exploited in a variety of biomedical applications, particularly in guidance of peripheral nerve regeneration.