Abstract
1. 1. Phospholipase C (phosphatidylcholine cholinephosphohydrolase, EC 3.1.4.3) has been purified 450-fold over the growth medium of Bacillus cereus, with overall recovery of 23%. 2. 2. Purification was performed using both (NH 4) 2SO 4 and ethanol precipitation, protamine sulfate fractionation, Sephadex G-100 gel filtration followed by ion exchange chromatography on DEAE- and CM-sephadex. 3. 3. The preparation appeared to be pure on disc electrophoresis at pH 2.3 in 6 M urea, whereas a molecular weight between 21 000 and 25 000 could be derived from gel filtration on Sephadex G-100. 4. 4. No hemolytic activity towards intact human erythrocytes could be established. Treatment of human red cell ghosts with pure phospholipase C resulted in a nearly complete degradation of the main phospholipid classes (except for sphingomyelin) up to 70% of total phosphorus. Similar results were obtained with liposomes derived from human erythrocyte total lipids.
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