Fifteen strains of filamentous fungi from the Culture Collection of Fungi (Charles University, Prague) were tested for their lipid production, fatty acid composition with emphasis on accumulation of oligounsaturated fatty acids. All cultures contained palmitic (16:0), palmitoleic (16:1), stearic (18:0), oleic (18:1), linoleic (18:2) and gamma-linolenic (18:3) acid (GLA). The mycelium of Cunninghamella elegans, Rhizopus arrhizus, Mortierella parvispora, M. elongata and M. alpina contained arachidonic acid (ARA) in the range of 2.3-33.5 % of the total fatty acids. The strains used in our experiment were capable to accumulate a relatively high amount of intracellular lipid (9.6-20.1 % in dry biomass). The highest content of GLA (22.3 mg/g) was found in Mucor circinelloides. The strain of M. alpina containing 47.1 mg/g of ARA could be considered as the best producer of ARA.