Composite films of graphene oxide (GO) sheets and silk fibroin (SF) with layered structures have been prepared by facile solution casting of SF-GO hydrogels. The as-prepared composite film containing 15% (by weight, wt%) of SF shows a high tensile strength of 221 ± 16 MPa and a failure strain of 1.8 ± 0.4%, which partially surpass those of natural nacre. Particularly, this composite film also has a high modulus of 17.2 ± 1.9 GPa. The high mechanical properties of this composite film can be attributed to its high content of GO (85 wt%), compact layered structure and the strong hydrogen bonding interaction between SF chains and GO sheets.