The enzymatic synthesis of phosphatidylserine and purification by CM-cellulose column chromatography

Abstract

Phosphatidylserine has been prepared from phosphatidylcholine by a one-step transphosphatidylation catalyzed by phospholipase D in the presence of L-serine. The resulting mixture of phosphatidylserine and phosphatidic acid is easily and rapidly separated by CM-cellulose column chromatography using step=wise elution with solvents containing increasing percentages of methanol in chloroform. The over-all yield of the procedure is 40-50% depending on the scale of the preparation. CM-Cellulose column chromatography proved to be extremely useful in separating phospholipid mixtures obtained by phosphatidyltransferase reactions of phospholipase D and is also suitable for fractionation of other lipid extracts.