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Biosynthetic preparation of selectively deuterated phosphatidylcholine in genetically modified Escherichia coli

Maric, Selma LU ; Thygesen, Mikkel B; Schiller, Jürgen; Marek, Magdalena; Moulin, Martine; Haertlein, Michael; Forsyth, V Trevor; Bogdanov, Mikhail; Dowhan, William and Arleth, Lise, et al. (2015) In Applied Microbiology and Biotechnology 99(1). p.54-241
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... (More)

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.

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publishing date
type
Contribution to journal
publication status
published
keywords
Culture Media/chemistry, Deuterium/metabolism, Escherichia coli/genetics, Metabolic Engineering, Metabolic Networks and Pathways/genetics, Phosphatidylcholines/metabolism, Staining and Labeling/methods
in
Applied Microbiology and Biotechnology
volume
99
issue
1
pages
14 pages
publisher
Springer
external identifiers
  • scopus:84925535388
ISSN
1432-0614
DOI
10.1007/s00253-014-6082-z
language
English
LU publication?
no
id
917d2199-f8a2-4c97-bedc-5ff26e746919
date added to LUP
2018-07-19 11:58:13
date last changed
2019-04-30 03:59:10
@article{917d2199-f8a2-4c97-bedc-5ff26e746919,
  abstract     = {<p>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. </p>},
  author       = {Maric, Selma and Thygesen, Mikkel B and Schiller, Jürgen and Marek, Magdalena and Moulin, Martine and Haertlein, Michael and Forsyth, V Trevor and Bogdanov, Mikhail and Dowhan, William and Arleth, Lise and Pomorski, Thomas Günther},
  issn         = {1432-0614},
  keyword      = {Culture Media/chemistry,Deuterium/metabolism,Escherichia coli/genetics,Metabolic Engineering,Metabolic Networks and Pathways/genetics,Phosphatidylcholines/metabolism,Staining and Labeling/methods},
  language     = {eng},
  number       = {1},
  pages        = {54--241},
  publisher    = {Springer},
  series       = {Applied Microbiology and Biotechnology},
  title        = {Biosynthetic preparation of selectively deuterated phosphatidylcholine in genetically modified Escherichia coli},
  url          = {http://dx.doi.org/10.1007/s00253-014-6082-z},
  volume       = {99},
  year         = {2015},
}