Purification of plasmalogens usingRhizopus delemar lipase andNaja naja naja phospholipase A₂

Abstract

Bovine heart ChoGpl (choline glycerophospholipid) and bovine brain EtnGpl (ethanolamine glycerophospholipid) contain diacyl, alkenylacyl and alkylacyl analogs. Purification of plasmalogens was achieved using R. delemar lipase and N. naja naja phospholipase A2 digestion. The R. delemar lipase hydrolyzes the acyl bond at the 1-position of 1,2-diacyl glycerophospholipids. The N. naja naja phospholipase A2 has greater activity with diacyl and alkylacyl than with alkenylacyl glycerophospholipids. These enzymes were mainly used to remove diacyl and alkylacyl analogs respectively. When the diacyl types were removed by double incubation with R. delemar lipase, the plasmalogen content was 94.2% +/- 0.21% (mean +/- S.E.M., n = 4) for PlsCho (plasmenylcholine) and 94.9% +/- 0.19% (mean +/- S.E.M., n = 3) for PlsEtn (plasmenylethanolamine). Recoveries were 74% and 88% respectively. These partially purified plasmalogens were treated with N. naja naja phospholipase A2. Finally, 97.7% +/- 0.24% (mean +/- S.E.M., n = 4) and 98.8% +/- 0.27% (mean +/- S.E.M., n = 3) pure plasmalogens were obtained for PlsCho and PlsEtn respectively. Plasmalogens were recovered in an overall yield of 7.7% +/- 0.7% (mean +/- S.E.M., n = 4) and 10.2% +/- 1.2% (mean +/- S.E.M., n = 3) for PlsCho and PlsEtn.

The hydrolysis of phosphatidylcholine by an immobilized lipase: Optimization of hydrolysis in organic solvents

“…Phospholipolytic activity has since been demonstrated in several purified lipases, including those from porcine pancrease (2,3), human liver (4) and Staphylococcus hyicus (5). This capability has been employed for the preparation of PLs and LPLs of defined structure (6,7} and for the removal of PL contaminants from preparations of plasmalogens (8,9).…”

Applications of lipase

“…"Slimming down," which is currently in vogue, can be carried out by fat removal by using lipase as a component of topical creams (76) or by its oral administration (77). Hirashima et al (78) carried out the hydrolysis of acyl bonds at the 1position of 1,2 diacylglycerophospholipids for the purification of plasmalogens with R. delemar lipase.…”