Biopolymer-based optical waveguides with low-loss light guiding performance and good biocompatibility are highly desired for applications in biomedical photonic devices. Herein, we report the preparation of silk optical fiber waveguides through bioinspired in situ mineralizing spinning, which possess excellent mechanical properties and low light loss. Natural silk fibroin was used as the main precursor for the wet spinning of the regenerated silk fibroin (RSF) fibers. Calcium carbonate nanocrystals (CaCO NCs) were in situ grown in the RSF network and served as nucleation templates for mineralization during the spinning, leading to the formation of strong and tough fibers. CaCO NCs can guide the structure transformation of silk fibroin from random coils to β-sheets, contributing to enhanced mechanical properties. The tensile strength and toughness of the obtained fibers are up to 0.83 ± 0.15 GPa and 181.98 ± 52.42 MJ·m, obviously higher than those of natural silkworm silks and even comparable to spider silks. We further investigated the performance of the fibers as optical waveguides and observed a low light loss of 0.46 dB·cm, which is much lower than natural silk fibers. We believed that these silk-based fibers with excellent mechanical and light propagation properties are promising for applications in biomedical light imaging and therapy.