24,25-Methylenecholest-5-en-3β-ol, which has not been reported previously to exist in nature, was isolated from mycelia of an arachidonic acid-producing fungus,Mortierella alpina 1S-4. Desmosterol, ergosta-5,24(25)-dien-3β-ol and ergosta-5,25-dien-3β-ol were also found in the fungus, but ergosterol and cholesterol were not detected.