Direct Enzymatic Synthesis of 1-Phosphatidyl-β-D-glucose by Engineered Phospholipase D

Arisa Inoue, Masaatsu Adachi, Jasmina Damnjanović, Hideo Nakano, Yugo Iwasaki

Research output: Contribution to journalArticlepeer-review

10 Citations (Scopus)

Abstract

A simple, direct method for the synthesis of 1-phosphatidyl-β-D-glucose (1-PGlc) was established, being the first example of direct enzymatic synthesis of this recently discovered, highly important bioactive phospholipid. The method employs phospholipase D (PLD, E.C. 3.1.4.4)-catalyzed transphosphatidylation, in which the polar head group of phosphatidylcholine is exchanged to glucose. Although wild-type PLD can catalyze the transfer reaction, it provides only 6-phosphatidyl-glucose, a positional isomer of 1-PGlc, due to its strong preference towards the primary hydroxyl group. To synthesize 1-PGlc, engineered PLD variants, previously isolated as the ones active on secondary hydroxyls of inositol, were screened for the ability to catalyze the reaction. One of the variants, 187K/191W/385Y (KWY), was identified as the best-performing in 1-PGlc synthesis, however, in addition to 1-PGlc the reaction with KWY also afforded undesired positional isomers as byproducts. To facilitate the isolation of 1-PGlc, the isomers were converted into the corresponding amines by reductive amination. Following the column chromatography, the desired product was isolated with the overall yield of 12.5 %. The structure of the product was confirmed by 1H-NMR analysis.

Original languageEnglish
Pages (from-to)4121-4125
Number of pages5
JournalChemistrySelect
Volume1
Issue number13
DOIs
Publication statusPublished - 2016 Aug 16
Externally publishedYes

Keywords

  • enzyme catalysis
  • phospholipase D, phosphatidylglucoside
  • phospholipid
  • transphosphatidylation

ASJC Scopus subject areas

  • Chemistry(all)

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