Infection caused by the fungus Cryptococcus neoformans is potentially fatal. A highly active extracellular phospholipase, demonstrating phospholipase B (PLB), lysophospholipase (LPL) and lysophospholipase\transacylase (LPTA) activities, was purified to homogeneity from C. neoformans using (NH % ) # SO % fractionation, and hydrophobic-interaction, anion-exchange and gel-filtration chromatography. All three enzyme activities copurified as a single protein with an apparent molecular mass of 70-90 kDa by SDS\PAGE and 160-180 kDa by gel filtration. The ratio of the three activities remained constant after each purification step. The amino acid composition, as well as the sequences of the N-terminus and of five internal peptide fragments were novel. The protein was an acidic glycoprotein containing Nlinked carbohydrate moieties, with pI values of 5.5 and 3.5. The apparent V max values for PLB and LPL activities were 12.3 and 870 µmol\min per mg of protein respectively ; the corresponding