Purification by affinity chromatography of phospholipase C from Bacillus cereus.

Phospholipase C (phosphatidylcholine cholinephosphohydrolase, EC 3.1.4.3) is a bacterial enzyme widely used in membrane and phospholipid studies. For such studies the highest possible degree of purity is required. Both the Bacillus cereus and the Clostridium perfringens enzymes have been purified to apparent homogeneity using conventional techniques [l-3]. However, using a phospholipase C-hyperproducing strain of B. cereus [4], homogeneous preparations of enzyme were not consistently obtained using the purification scheme described [2] (C. Little and A. B. Otnaess, unpublished observation). The C7. perfringens enzyme has also been purified by a method involving affinity chromatography on agarose-linked egg yolk lipoprotein [ 51 and we have applied this technique to the purification of the B. cereus enzyme. The purification scheme here presented is simpler and more convenient and gives higher yields of enzyme than do the previously published methods.