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. 2020 Aug 26;5(35):22395-22401.
doi: 10.1021/acsomega.0c02823. eCollection 2020 Sep 8.

Enzyme-Assisted Synthesis of High-Purity, Chain-Deuterated 1-Palmitoyl-2-oleoyl- sn-glycero-3-phosphocholine

Oliver Bogojevic  1 , Anna E Leung  1
Affiliations

Enzyme-Assisted Synthesis of High-Purity, Chain-Deuterated 1-Palmitoyl-2-oleoyl- sn-glycero-3-phosphocholine

Oliver Bogojevic et al. ACS Omega. .

Abstract

1-Palmitoyl-d 31-2-oleoyl-d 32-sn-glycero-3-phosphocholine (POPC-d 63) with the palmitoyl and oleoyl chains deuterium-labeled was produced in three steps from 1-palmitoyl-2-hydroxy-sn-glycero-3-phosphocholine, deuterated palmitic acid, and deuterated oleic anhydride. Esterification at the sn-2 position was achieved under standard chemical conditions, using DMAP to catalyze the reaction between the 2-lysolipid and oleic anhydride-d 64. Complete regioselective sn-1 acyl substitution was achieved in two steps using operationally simple, enzyme-catalyzed regioselective hydrolysis and esterification to substitute the sn-1 chain for a perdeuterated analogue. This method provides chain-deuterated POPC with high chemical purity (>96%) and complete regiopurity, useful for a variety of experimental techniques. This chemoenzymatic semisynthetic approach is a general, modular method of producing highly pure, mixed-acyl phospholipids, where the advantages of both chemical synthesis (efficiency, high yields) and biocatalytic synthesis (specificity, nontoxicity) are realized.

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Conflict of interest statement

The authors declare no competing financial interest.

Figures

Figure 1
Figure 1
Chain-deuterated isotopologue of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC-d63), with stereospecific numbering indicated.
Scheme 1
Scheme 1. Proposed Chemoenzymatic Synthesis of Chain-Deuterated POPC
Scheme 2
Scheme 2. Synthesis of POPC-d63 and Rationalization for the Presence of Isotopologues of DPPC
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