Purification of plasmalogens using Rhizopus delemar lipase and Naja naja naja phospholipase A 2

Bovine heart ChoGpl (choline glycerophospholipid) and bovine brain EtnGpl (ethanolamine glycerophospholipid) contain diacyl, alkenylacyl and alkylacyl analogs. Purification of plasmalogens was achieved usingR. delemar lipase andN. naja naja phospholipase A2 digestion. TheR. delemar lipase hydrolyzes the acyl bond at the 1-position of 1,2-diacyl glycerophospholipids. TheN. 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 withR. 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 withN. 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.