Chitin biopolymer production and its by-product chitosan show great potential. These biomaterials have great applicability in various fields because they are non-toxic, biodegradable, biocompatible, and have antimicrobial effects. The most common source of chitin and chitosan is the crustaceous shell; however, mushrooms are an alternative source for isolating these biopolymers because their cellular wall has a high content of chitin, which may be transformed into chitosan through a deacetylation reaction. The main objective of this research was to obtain chitosan through the deacetylation of chitin isolated from the Ganoderma lucidum basidiomycetes mushroom, which is obtained through biotechnological culture. The material characterization was performed using X-ray diffraction, Fourier transform infrared spectroscopy, thermogravimetric analysis, and an evaluation of cytotoxicity comparing the results obtained with results for commercial chitosan. Protocol results showed that chitosan obtained from this mushroom had a significant similitude with commercial chitosan, yet the one obtained using P2 protocol was the one that rendered the best results: including diffractogram peaks, characteristic infrared analysis bands, and an 80.29 % degree of deacetylation. Cytotoxicity in vitro testing showed that the material was non-toxic; furthermore, it rendered very promising information regarding the evaluation of future applications of this biomaterial in the field of biomedicine.