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This study investigates the mycelial biomass production and chitosan extraction potential of various Basidiomycota strains, including <i>Heterobasidion annosum</i>, <i>Phanerochaete chrysosporium</i>, <i>Pleurotus ostreatus</i>, <i>Trametes versicolor</i>, and <i>Lentinus lepideus.</i> Both submerged fermentation (SF) and solid-state fermentation (SSF) methods were employed. The chitosan yield in basidiocarps of <i>Pleurotus ostreatus</i>, <i>Agaricus bisporus</i>, and <i>Ganoderma applanatum</i> 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, <i>P. chrysosporium</i> exhibited the highest mycelial biomass concentration in SF (1.03 g 100 mL^–1) after 14 days, while <i>T. versicolor</i> achieved the highest biomass concentration in SSF (3.65 g 100 mL^–1). The highest chitosan yield was obtained from the mycelium of <i>P. chrysosporium</i> (0.38%) and <i>T. versicolor</i> (0.37%) in shaken SF. Additionally, commercially cultivated <i>A. bisporus</i> 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.