Biosynthetic preparation of selectively deuterated phosphatidylcholine in genetically modified Escherichia coli
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Biosynthetic preparation of selectively deuterated phosphatidylcholine in genetically modified Escherichia coli. / Maric, Selma; Thygesen, Mikkel Boas; Schiller, Jürgen; Marek, Magdalena; Moulin, Martine; Haertlein, Michael; Forsyth, V. Trevor; Bogdanov, Mikhail; Dowhan, William; Arleth, Lise; Günther-Pomorski, Thomas.
I: Applied Microbiology and Biotechnology, Bind 99, Nr. 1, 2015, s. 241-254.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - Biosynthetic preparation of selectively deuterated phosphatidylcholine in genetically modified Escherichia coli
AU - Maric, Selma
AU - Thygesen, Mikkel Boas
AU - Schiller, Jürgen
AU - Marek, Magdalena
AU - Moulin, Martine
AU - Haertlein, Michael
AU - Forsyth, V. Trevor
AU - Bogdanov, Mikhail
AU - Dowhan, William
AU - Arleth, Lise
AU - Günther-Pomorski, Thomas
PY - 2015
Y1 - 2015
N2 - 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.
AB - 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.
KW - Biosynthesis
KW - E. coli
KW - Evolution in the test tube
KW - Glycerophospholipids
KW - Mass spectrometry
KW - Neutron scattering
KW - NMR
KW - Phosphatidylcholine
KW - Selective deuteration
U2 - 10.1007/s00253-014-6082-z
DO - 10.1007/s00253-014-6082-z
M3 - Journal article
C2 - 25301578
VL - 99
SP - 241
EP - 254
JO - Applied Microbiology and Biotechnology
JF - Applied Microbiology and Biotechnology
SN - 0175-7598
IS - 1
ER -
ID: 130324912