means of obtaining very high yields of 1-acyl-rac-glycero-3-phosphate (LPA1) was
to carry out the reaction at a temperature below the melting point of this product.
Precipitation of LPA1 occurred in the reaction mixture, and close to quantitative
yields were obtained with a number of different fatty acids. The temperature re-
quired to achieve good results depended on the chain length and degree of unsatura-
tion of the fatty acid.
The reaction conditions and yields in the published methods for enzymatic LPA
synthesis are summarized in Table 1.
Acylation of glycerophosphorylcholine and related substances
Acid anhydrides are useful acyl donors in some lipase-catalyzed reactions. A high
yield (71 %) of lysophosphatidylcholine (LPC) was obtained in the acylation of
glycerophosphorylcholine using Lipozyme as catalyst and methylene chloride as
solvent. Glycerophosphorylcholine was not dissolved in the solvent, but acylation
of the substrate suspension proved to be quite effective (Mazur et al., 1991). The
reaction conditions and yields in the published methods for enzymatic LPC synth-
esis are summarized in Table 1.
Glycerophosphorylcholine was acylated using vinyl esters (Virto and Adlercreutz,
2000a) in a manner similar to the acylation of glycerophosphate described above. In
this case,Candida antarcticalipase B was shown to be a suitable catalyst. It was
possible to carry out the reaction in a solvent-free system, but higher yields were
obtained when a small amount oft-butanol (about the same weight as the sub-
14.5 Esterification reactions for the incorporation of fatty acids into lipids 299
Table 1.Lipase-catalyzed acylation of glycerophosphate and glycerophosphorylcholine.
Enzyme Solvent Acyl
donor
Product^1 Yield^2 Comments ReferenceRhizomucor
mieheilipase^3
No Fatty
acidLPA 32 %
(isolated)Han and Rhee, 1995Rhizomucor
mieheilipase^3
No Fatty
acidLPA 45 % Water activity
controlHan and Rhee, 1998Rhizopus arrhizus
lipase on
polypropylene
No Vinyl
estersLPA >90 % Product
precipitationVirto et al, 1999Rhizomucor
mieheilipase^3
CH 2 Cl 2 Acid
anhydrideLPC 71 %
(isolated)Mazur et al., 1991Rhizomucor
mieheilipase^3
No Fatty
acidLPC 36 % Water activity
controlHan and Rhee, 1998Candida
antarctica
lipase B^4
t-BuOH Vinyl
estersLPC >95 % Product
precipitationVirto and
Adlercreutz, 2000a(^1) LPA, lysophosphatidic acid; LPC, lysophosphatidylcholine.
(^2) Analytical yields unless otherwise stated.
(^3) Lipozyme, commercially available immobilized preparation.
(^4) Novozyme 435, commercially available immobilized preparation.