title: Biosynthetic preparation of selectively deuterated phosphatidylcholine in genetically modified Escherichia coli.
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.
authors.affiliation: Structural Biophysics, Niels Bohr Institute, Faculty of Science, University of Copenhagen, Universitetsparken 5, 2100, Copenhagen, Denmark.
authors.forename: Selma
authors.initials: S
authors.lastname: Maric
authors.forename: Mikkel B
authors.initials: MB
authors.lastname: Thygesen
authors.forename: Jürgen
authors.initials: J
authors.lastname: Schiller
authors.forename: Magdalena
authors.initials: M
authors.lastname: Marek
authors.forename: Martine
authors.initials: M
authors.lastname: Moulin
authors.forename: Michael
authors.initials: M
authors.lastname: Haertlein
authors.forename: V Trevor
authors.initials: VT
authors.lastname: Forsyth
authors.forename: Mikhail
authors.initials: M
authors.lastname: Bogdanov
authors.forename: William
authors.initials: W
authors.lastname: Dowhan
authors.forename: Lise
authors.initials: L
authors.lastname: Arleth
authors.forename: Thomas Günther
authors.initials: TG
authors.lastname: Pomorski
mesh_terms: D003470:Culture Media / Q000737:chemistry; D003903:Deuterium / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D060847:Metabolic Engineering; D053858:Metabolic Networks and Pathways / Q000235:genetics*; D010713:Phosphatidylcholines / Q000378:metabolism*; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D003470:Culture Media; D010713:Phosphatidylcholines; D003903:Deuterium