Biosynthetic preparation of selectively deuterated phosphatidylcholine in genetically modified Escherichia coli

Research output: Contribution to journalArticle

Abstract

Phosphatidylcholine (PC) is a major component of eukaryotic cell membranes and one of the most commonly used phospholipids for reconstitution of membrane proteins into carrier systems such as lipid vesicles, micelles and nanodiscs. Selectively deuterated versions of this lipid have many applications, especially in structural studies using techniques such as NMR, neutron reflectivity and small-angle neutron scattering. Here we present a comprehensive study of selective deuteration of phosphatidylcholine through biosynthesis in a genetically modified strain of Escherichia coli. By carefully tuning the deuteration level in E. coli growth media and varying the deuteration of supplemented carbon sources, we show that it is possible to achieve a controlled deuteration for three distinct parts of the PC lipid molecule, namely the (a) lipid head group, (b) glycerol backbone and (c) fatty acyl tail. This biosynthetic approach paves the way for the synthesis of specifically deuterated, physiologically relevant phospholipid species which remain difficult to obtain through standard chemical synthesis.

Details

Authors
  • Selma Maric
  • Mikkel B Thygesen
  • Jürgen Schiller
  • Magdalena Marek
  • Martine Moulin
  • Michael Haertlein
  • V Trevor Forsyth
  • Mikhail Bogdanov
  • William Dowhan
  • Lise Arleth
  • Thomas Günther Pomorski
External organisations
  • University of Copenhagen
Research areas and keywords

Keywords

  • Culture Media/chemistry, Deuterium/metabolism, Escherichia coli/genetics, Metabolic Engineering, Metabolic Networks and Pathways/genetics, Phosphatidylcholines/metabolism, Staining and Labeling/methods
Original languageEnglish
Pages (from-to)241-54
Number of pages14
JournalApplied Microbiology and Biotechnology
Volume99
Issue number1
Publication statusPublished - 2015 Jan
Publication categoryResearch
Peer-reviewedYes
Externally publishedYes