Enzymatic hydrolysis by bacterial phospolipases C and D of immobilized radioactive sphingomyelin and phosphatidylcholine.
An assay system for phospholipases C has been described with sphingomyelin immobilized to octyl-Sepharose CL-4B as substrate. The immobilization procedure was further developed and used with [14 C-choline]-sphingomyelin and [14C-choline] phosphatidylcholine (lecithin). These immobilized radioactive phospholipids made the enzymatic assays easier to perform and made it possible to increase the sensitivity. Furthermore, since release of the choline part instead of the phosphate part of the substrate molecule was measured, it was possible to use this assay for phospholipase D as well. The enzyme characteristics of phospholipase D from Corynebacterium ovis were compared in this test system with those of three phospholipases C (from Clostridium perfringens, Bacillus cereus and Staphylococcus aureus) with respect to hydrolysing capacities and optimal ion concentrations.
D001409:Bacillus cereus / Q000201:enzymology; D001419:Bacteria / Q000201:enzymology*; D002413:Cations, Divalent / Q000494:pharmacology; D003016:Clostridium perfringens / Q000201:enzymology; D003352:Corynebacterium / Q000201:enzymology; D010713:Phosphatidylcholines / Q000378:metabolism*; D010739:Phospholipase D / Q000378:metabolism*; D010740:Phospholipases / Q000378:metabolism*; D013109:Sphingomyelins / Q000378:metabolism*; D013211:Staphylococcus aureus / Q000201:enzymology; D010738:Type C Phospholipases / Q000378:metabolism*
D002413:Cations, Divalent; D010713:Phosphatidylcholines; D013109:Sphingomyelins; D010740:Phospholipases; D010738:Type C Phospholipases; D010739:Phospholipase D