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This study investigates the mycelial biomass production and chitosan extraction potential of various Basidiomycota strains, including Heterobasidion annosum, Phanerochaete chrysosporium, Pleurotus ostreatus, Trametes versicolor, and Lentinus lepideus. Both submerged fermentation (SF) and solid-state fermentation (SSF) methods were employed. The chitosan yield in basidiocarps of Pleurotus ostreatus, Agaricus bisporus, and Ganoderma applanatum was also evaluated as a reference material. The chitosan extracted from fungal cells was characterized using elemental analyses and FTIR spectroscopy. Among the cultivated strains, P. chrysosporium exhibited the highest mycelial biomass concentration in SF (1.03 g 100 mL–1) after 14 days, while T. versicolor achieved the highest biomass concentration in SSF (3.65 g 100 mL–1). The highest chitosan yield was obtained from the mycelium of P. chrysosporium (0.38%) and T. versicolor (0.37%) in shaken SF. Additionally, commercially cultivated A. bisporus demonstrated the highest chitosan yield in fungal fruiting bodies (1.7%). The extracted chitosan holds potential as a functional biopolymer additive for eco-friendly materials, serving as an alternative to synthetic wet and dry strength agents in packaging materials. [ABSTRACT FROM AUTHOR]