Fungal biomass of Rhizopus delemar, grown on bread waste, was used to develop filaments. The fungal cell wall materials were isolated from the biomass via alkali treatment. This material, which contains chitin and chitosan, was then used to develop a hydrogel serving as the spinning dope. Fungal filaments were produced via dry gel spinning using vanillin as a crosslinker in combination with glycerol as a plasticizer. The influence of both vanillin and glycerol on filament properties was evaluated through measurements of optical properties, swelling ratio, tenacity, initial modulus, and elongation at break. The filaments were characterized using swelling test, surface colour measurement, tensile tester and Fourier trans-form infrared spectrum (FT-IR). The results demonstrated that vanillin as a crosslinker enhanced the mechanical properties of fungal filaments, with the tenacity increasing by 20.5% compared to non-crosslinked filaments. However, the incorporation of 5% w/w glycerol led to a 42.9% reduction in tenacity relative to filaments without glycerol. Thermal treatment of crosslinked fungal filaments had negligible impact on tenacity but increased brittleness, limiting their processability. Microscopic images revealed the presence of dark spots on the monofilament surface, which may negatively influence the visual uniformity of resulting knitted fabrics. Additionally, filaments containing 5% w/w glycerol were able to be knitted; however, their multifilament yarns exhibited a tendency to break during the knit-ting process. Fungal monofilaments with tenacities of ~2.0–2.5cN/tex were twisted into multifilament yarns using 3–4 monofilaments. These yarns were successfully manufactured and demonstrated textile processability in knitting processes.