Hog pancreas lipase was covalently attached to the beads of poly(vinyl alcohol)-cryogel - a macroporous hydrogel prepared by means of freeze-thaw technique. The immobilized biocatalyst thus obtained was examined in the reaction of enantioselective hydrolysis of the ethyl ester of N-benzylidene derivative of DL-phehylalanine in the medium of acetonitrile (contained 5 vol.% of water without any buffers). Eighty-three %-enantiomeric excess of the L-amino acid was reached after 144 h. Virtually the same result was obtained in the repeated use of the same immobilized biocatalyst after its 6-months-storing in a refrigerator.