n-Hexadecylphosphonocholine → hexadecylphospho-L-serine

HRID 465962Notes

Reaction equation

HRID 465962 reaction equation
[NH2:1][C@H:2]([C:5]([OH:7])=[O:6])[CH2:3][OH:4].[CH2:8]([O:24][P:25](OCC[N+](C)(C)C)([OH:27])=[O:26])[CH2:9][CH2:10][CH2:11][CH2:12][CH2:13][CH2:14][CH2:15][CH2:16][CH2:17][CH2:18][CH2:19][CH2:20][CH2:21][CH2:22][CH3:23].C(OCC)C.C(Cl)(Cl)Cl.C(N(CC(O)=O)CC(O)=O)CN(CC(O)=O)CC(O)=O>C([O-])(=O)C.[Cl-].[Cl-].[Ca+2]>[CH2:8]([O:24][P:25]([O:4][CH2:3][C@@H:2]([C:5]([OH:7])=[O:6])[NH2:1])([OH:27])=[O:26])[CH2:9][CH2:10][CH2:11][CH2:12][CH2:13][CH2:14][CH2:15][CH2:16][CH2:17][CH2:18][CH2:19][CH2:20][CH2:21][CH2:22][CH3:23]

Participants

7

Reactant 4

HCID 3600n-HexadecylphosphonocholineHCID 5951L-SerineHCID 6049Ethylenediaminetetraacetic AcidHCID 21911965Chloroform ether

Product 1

HCID 10047119hexadecylphospho-L-serineYield 16.3%

Reagents, catalysts & solvents 2

HCID 175AcetateSolventHCID 5284359Calcium ChlorideSolvent

Conditions

Temperature
45 °C

Procedure

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.

Workup

  1. customat 45° C
  2. customhas been prepared from white cabbage
  3. additionare added to this solution
  4. temperatureAfter cooling to room temperature
  5. customthe organic solvents are removed
  6. stirringThe aqueous phase which remains is stirred with 4.3 times its volume of chloroform/methanol (5:8, v/v) for 30 minutes
  7. filtrationthe undissolved material (L-serine) is filtered off with suction
  8. addition1 volume of water and 3.7 volumes of chloroform are added to the filtrate
  9. stirringthe mixture is shaken for 10 minutes
  10. customthe organic phase is separated off
  11. concentrationconcentrated in vacuo
  12. customthe residue obtained
  13. customis separated by column chromatography on carboxymethyl cellulose (CM 52 Whatman, Na+ form) Elution

Sources & evidence

ORD record
ord-532b00d8952e4c378f6af736495ebc10
Source dataset
uspto-grants-1992_10
Patent number
US05155099

Reaction notes

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