Hydrolysis by phospholipase D of phospholipids in solution state or adsorbed on a silica matrix.

Phospholipases D (PLD) catalyse hydrolysis and transphosphatidylation reactions in phospholipids. In the present study, the hydrolytic activity for cabbage PLD was investigated with five different substrates (dipalmitoylphosphatidylethanolamine (DPPE), dipalmitoylphosphatidylcholine (DPPC), didecanoylphosphatidylcholine (DDPC), 1-alkyl-2-acetyl-sn-glycero-3-phosphocholine and lyso-phosphatidylcholine (lyso-PC)) in solution or adsorbed on a silica matrix. In the specific buffer solutions, where the substrates were proved to form large multilamellar polydisperse aggregates, PLD showed preference for DPPC > DPPE > DDPC > 1-alkyl-2-acetyl-sn-glycero-3-phosphocholine > lyso-PC. When the substrates were adsorbed on the silica matrix, PLD hydrolysed 1-alkyl-2-acetyl-sn-glycero-3-phosphocholine and lyso-PC, DDPC, but not DPPC or DPPE. Theoretical studies of the simplest possible adducts between the phospholipids and the silica matrix were performed. Examination of local geometries of DPPC showed a significant blocking of the P-O-X bond-prone to hydrolysis, which could possibly block the access of PLD. Immobilization of phospholipids could be applied for improving the yield of reactions catalysed by PLD as well as for performing a targeted production of short-chain length phosphatidic acid analogs.