Applied Microbiology and Biotechnology

, Volume 99, Issue 1, pp 241–254 | Cite as

Biosynthetic preparation of selectively deuterated phosphatidylcholine in genetically modified Escherichia coli

  • 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
Biotechnological products and process engineering

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.

Keywords

Glycerophospholipids Phosphatidylcholine Selective deuteration Biosynthesis E. coli Neutron scattering NMR Mass spectrometry Evolution in the test tube 

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Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Selma Maric
    • 1
    • 2
  • Mikkel B. Thygesen
    • 3
  • Jürgen Schiller
    • 4
  • Magdalena Marek
    • 2
  • Martine Moulin
    • 5
    • 6
  • Michael Haertlein
    • 5
  • V. Trevor Forsyth
    • 5
    • 6
  • Mikhail Bogdanov
    • 7
  • William Dowhan
    • 7
  • Lise Arleth
    • 1
  • Thomas Günther Pomorski
    • 2
  1. 1.Structural Biophysics, Niels Bohr Institute, Faculty of ScienceUniversity of CopenhagenCopenhagenDenmark
  2. 2.Center for Membrane Pumps in Cells and Disease, Department of Plant and Environmental Sciences, Faculty of ScienceUniversity of CopenhagenFrederiksberg CDenmark
  3. 3.CARB Centre, Department of Chemistry, Faculty of ScienceUniversity of CopenhagenFrederiksberg CDenmark
  4. 4.Institut für Medizinische Physik und Biophysik, Medizinische FakultätUniversität LeipzigLeipzigGermany
  5. 5.Life Sciences GroupInstitut Laue LangevinGrenobleFrance
  6. 6.Faculty of Natural Sciences & Institute for Science and Technology in MedicineKeele UniversityStaffordshireUK
  7. 7.Department of Biochemistry and Molecular BiologyUniversity of Texas Medical School at HoustonHoustonUSA

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