Aqueous silk fibroin (SF) sol is a colloidal solution. With the colloidal hydration layer and electrostatic repulsion, the SF sol can hardly make the efficient collision/assembly among micelles and perform like a following sol for a long time. In this paper, hydrophilic silk-based sequences (HSF) derived from SF molecules were obtained by immersing the dried SF condensates with water and extracting the dissolving fraction. The HSF was obtained by immersing the SF condensate dried at the temperature of [Formula: see text]–[Formula: see text] and relative humid of 55–60% in water and collected the lixivium. The dissolving ratio was about 30%. The HSF sol (0.5%, w/v) self-assembled into the mesoscopic 3D nanofibrous network within 8 h. The obtained HSF nanofibers were 10–100 [Formula: see text] in length and 50–100 nm in diameter. The HSF nanofiber possesses similar hierarchical structure consisting of nanofibrils bundles to the native silk fiber. There were significant aggregation structure transitions from random coil to [Formula: see text]-sheet and amorphous chains to Silk II crystal aggregation during the formation of HSF nanofibers. The HSF nanofiber holds the potential to give further insight into the reconstruction of native silk in vitro and the fabrication of tough silk-based biomaterials.
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