Production of chitin deacetylases from the phytopathogenic fungus Colletotrichum gloeosporioides was successfully achieved by submerged fermentation. The highest specific activity of 0.018Umg⁻¹ of protein was obtained after 96h of cultivation at pH 6 and 28°C. Two bands with molecular weights of 35kDa and 170kDa determined with SDS-PAGE displayed deacetylase activities as detected in the zymograms. Reacetylated commercial chitosan (52% acetylation degree) was used as substrate for the extracellular crude extract in order to estimate the kinetic parameters of acetate production as undirected deacetylation measurement. The highest acetate production of 12.8μmolmL⁻¹ was obtained using 7.5mgmL⁻¹ of substrate. The produced enzyme from C. gloeosporioides achieved up to 25% deacetylation of a chitin substrate (hydrolyzed biological chitin) having 80% degree of acetylation, M_W of 102×10³gmol⁻¹ and a crystallinity index of ca. 60%.