Silk fibroin (SF)-based materials are characterized by their outstanding biocompatibility and biodegradability and are considered as the most promising candidates for next-generation flexible electronics. In order to generate such devices, SF can be mixed with carbon nanotubes (CNTs) which feature excellent mechanical, electrical, and thermal properties. However, obtaining regenerated SF with homogeneous dispersion of CNTs in a sustainable manner represents a challenging task, mainly due to the difficulty in overcoming van der Waals forces and strong π-π interactions that hold together the CNT structure. In this study, a one-pot strategy for fabricating SF/CNT films is proposed by designing SF as a modifier of CNTs through non-covalent interactions with the assistance of aqueous phosphoric acid solution. Glycerol (GL) was introduced, endowing the SF/GL/CNT composite film with excellent flexibility and stretchability. The sustainable strategy greatly simplifies the preparation process, avoiding dialysis of SF and the use of artificial dispersants. The as-fabricated SF/GL/CNT films showed an excellent mechanical strength of 1.20 MPa and high sensitivity with a gauge factor of up to 13.7 toward tensile deformation. The composite films had a sensitive monitoring capability for small strains with detection limits as low as 1% and can be assembled into versatile sensors to detect human movement. Simultaneously, the composite films showed a superb thermosensitive capacity (1.64% °C-1), which satisfied the requirement of real-time and continuous skin temperature monitoring. We anticipate that the presented one-pot strategy and the prepared composite films could open a new avenue for forthcoming technologies for electronic skins, personal health monitoring, and wearable electronics.