PURIFICATION AND SOME PROPERTIES OF PHOSPHOLIPASE-B FROM SCHIZOSACCHAROMYCES-POMBE

Phospholipase B from Schizosaccharomyces pombe was purified by ammoniu m sulfate fractionation and chromatographed on phenyl-Sepharose CL-4B, DEAE-Toyopearl 650M, and TSK gel G4000SW columns, The purified enzyme was a glycoprotein with molecular weight of approximately 300,000 and 100,000-150,000 by gel filtration and SDS-polyacrylamide gel electrop horesis, respectively, The isoelectric point was pH 4.7, The optimum p H of the enzyme was 2.5 and no activity was detected at neutral and al kaline pHs, The enzyme was not heat-stable, Enzyme activity was slight ly stimulated by divalent ions except Fe2+ and 0.1% sodium deoxycholat e, and inhibited by Fe2+, Fe3+, 0.1% sodium dodecyl sulfate, and 0.01% cetyltrimethylammonium bromide. The enzyme hydrolyzed mono- and diacy lphospholipids, and phosphatidylinositol was hydrolyzed most preferent ially, Triglyceride was not hydrolyzed, The enzyme also had acyltransf erase activity on lysophosphatidylcholine, forming the corresponding d iacylphosphatidylcholine.