Biosynthetic preparation of selectively deuterated phosphatidylcholine in genetically modified Escherichia coli

Selma Maric; Mikkel Boas Thygesen; Jürgen Schiller; Magdalena Marek; Martine Moulin; Michael Haertlein; V. Trevor Forsyth; Mikhail Bogdanov; William Dowhan; Lise Arleth; Thomas Günther-Pomorski

doi:10.1007/s00253-014-6082-z

Biosynthetic preparation of selectively deuterated phosphatidylcholine in genetically modified Escherichia coli

Selma Maric, Mikkel Boas Thygesen, Jürgen Schiller, Magdalena Marek, Martine Moulin, Michael Haertlein, V. Trevor Forsyth, Mikhail Bogdanov, William Dowhan, Lise Arleth, Thomas Günther-Pomorski^*

^*Corresponding author for this work

23 Citations (Scopus)

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.

Original language	English
Journal	Applied Microbiology and Biotechnology
Volume	99
Issue number	1
Pages (from-to)	241-254
Number of pages	14
ISSN	0175-7598
DOIs	https://doi.org/10.1007/s00253-014-6082-z
Publication status	Published - Jan 2014

Keywords

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

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Maric, S., Thygesen, M. B., Schiller, J., Marek, M., Moulin, M., Haertlein, M., Forsyth, V. T., Bogdanov, M., Dowhan, W., Arleth, L., & Günther-Pomorski, T. (2014). Biosynthetic preparation of selectively deuterated phosphatidylcholine in genetically modified Escherichia coli. Applied Microbiology and Biotechnology, 99(1), 241-254. https://doi.org/10.1007/s00253-014-6082-z

Maric, S, Thygesen, MB, Schiller, J, Marek, M, Moulin, M, Haertlein, M, Forsyth, VT, Bogdanov, M, Dowhan, W, Arleth, L & Günther-Pomorski, T 2014, 'Biosynthetic preparation of selectively deuterated phosphatidylcholine in genetically modified Escherichia coli', Applied Microbiology and Biotechnology, vol. 99, no. 1, pp. 241-254. https://doi.org/10.1007/s00253-014-6082-z

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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.",

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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 - 2014/1

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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

SN - 0175-7598

VL - 99

SP - 241

EP - 254

JO - Applied Microbiology and Biotechnology

JF - Applied Microbiology and Biotechnology

IS - 1

ER -

Biosynthetic preparation of selectively deuterated phosphatidylcholine in genetically modified Escherichia coli

Abstract

Keywords

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