The isolation and potential applications of chitosan from the fungal source is an alternative to crustacean chitosan and synthetic polymers. The strains of Basidiomycota (Heterobasidion annosum, Phanerochaete chrysosporium, Pleurotus ostreatus, Trametes versicolor, Lentinus lepideus) with different environmental distribution, ecological importance, and industrial application were studied for mycelial biomass production in submerged (SF) and solid state fermentation (SSF). Further, the chitosan from fungal cells was extracted and characterized by elemental analyses (nitrogen) and FTIR spectroscopy. The fruiting bodies of Pleurotus ostreatus, Agaricus bisporus and Ganoderma ap-planatum were chosen as a reference material of chitosan content in basidiocarps. Stationary SF gave the lowest biomass yield in comparison with shaken SF and SSF. The role of aeration and agitation in biomass production was clearly observed in shaken SF with the highest yield achieved by P. chrysosporium. The highest biomass yield in SSF produced P. ostreatus and T. versi-color. The content of chitosan within fungal species varied depending on the method of cultiva-tion. The highest chitosan yield among the cultivated strains was obtained from mycelium of P. chrysosporium and T. versicolor in SF. The highest chitosan yield in fungal fruiting bodies demon-strated commercially cultivated mushroom A. bisporus. The extracted chitosan is proposed as functional biopolymer additive for ecological materials to substitute the synthetic wet and dry strength agents in packaging materials.