The aim of this work was to increase access and activity of lipases bounded to subcellular structures of Rhizomucor miehei and Yarrowia lipolytica. To obtain this goal biomass of fungi was treated with digitonin and cetyltrimethylammo- nium bromide (СТАВ). The highest activity of endolipases was found for biomass incubated at 30°C for 30 min and 0.2% digitonin for R. miehei and 0.2% СТАВ for Y. lipolytica. The lipolytic activity of permeabilized cells for both strains was about 2,5 times higher compared with native cells.