n-Hexadecylphosphonocholine → hexadecylphospho-L-serine

Participants

Reaction Conditions

Reaction Steps

0.8 g of L-serine (7.6 mmol) is dissolved in 1.52 ml of 0.1M acetate buffer (pH 5.6), which is 0.09M in CaCl2, at 45° C. 31 mg (0.075 mmol) of n-hexadecylphosphonocholine, 1.6 ml of diethyl ether/chloroform (9:1, v/v, ethanol-free) and 200 mg of a phospholipase D enzyme preparation which has been prepared from white cabbage and has an activity of 0.9 U/ml (1 unit (U) converts 1 μmol of substrate per minute at 27° C.) in the reaction mixture, are added to this solution. The mixture is stirred at 45° C. for 2.2 hours. After cooling to room temperature, 3.31 ml of 0.1M EDTA solution are added and the organic solvents are removed by passing in nitrogen. The aqueous phase which remains is stirred with 4.3 times its volume of chloroform/methanol (5:8, v/v) for 30 minutes and the undissolved material (L-serine) is filtered off with suction. 1 volume of water and 3.7 volumes of chloroform are added to the filtrate, the mixture is shaken for 10 minutes, the organic phase is separated off and concentrated in vacuo, and the residue obtained is separated by column chromatography on carboxymethyl cellulose (CM 52 Whatman, Na+ form) Elution is carried out successively with 75 ml of chloroform, 300 ml each of chloroform/methanol 9:1, 8:2 and 7:3, then 1,750 ml of chloroform/methanol 1:1 (in each case v/v). The fraction size is 50 ml. 5 mg of pure n-hexadecylphosphono-L-serine (I, R=C16H33) are obtained from fractions 20-39.

Provenance & Evidence