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 B; Schiller, Jürgen; Marek, Magdalena; Moulin, Martine; Haertlein, Michael; Forsyth, V Trevor; Bogdanov, Mikhail; Dowhan, William; Arleth, Lise; Pomorski, Thomas Günther.

In: Applied Microbiology and Biotechnology, Vol. 99, No. 1, 01.2015, p. 241-54.

Research output: Contribution to journalArticle

Harvard

Maric, S, Thygesen, MB, Schiller, J, Marek, M, Moulin, M, Haertlein, M, Forsyth, VT, Bogdanov, M, Dowhan, W, Arleth, L & Pomorski, TG 2015, 'Biosynthetic preparation of selectively deuterated phosphatidylcholine in genetically modified Escherichia coli', Applied Microbiology and Biotechnology, vol. 99, no. 1, pp. 241-54. https://doi.org/10.1007/s00253-014-6082-z

APA

Maric, S., Thygesen, M. B., Schiller, J., Marek, M., Moulin, M., Haertlein, M., Forsyth, V. T., Bogdanov, M., Dowhan, W., Arleth, L., & Pomorski, T. G. (2015). Biosynthetic preparation of selectively deuterated phosphatidylcholine in genetically modified Escherichia coli. Applied Microbiology and Biotechnology, 99(1), 241-54. https://doi.org/10.1007/s00253-014-6082-z

CBE

Maric S, Thygesen MB, Schiller J, Marek M, Moulin M, Haertlein M, Forsyth VT, Bogdanov M, Dowhan W, Arleth L, Pomorski TG. 2015. Biosynthetic preparation of selectively deuterated phosphatidylcholine in genetically modified Escherichia coli. Applied Microbiology and Biotechnology. 99(1):241-54. https://doi.org/10.1007/s00253-014-6082-z

MLA

Vancouver

Author

Maric, Selma ; Thygesen, Mikkel B ; Schiller, Jürgen ; Marek, Magdalena ; Moulin, Martine ; Haertlein, Michael ; Forsyth, V Trevor ; Bogdanov, Mikhail ; Dowhan, William ; Arleth, Lise ; Pomorski, Thomas Günther. / Biosynthetic preparation of selectively deuterated phosphatidylcholine in genetically modified Escherichia coli. In: Applied Microbiology and Biotechnology. 2015 ; Vol. 99, No. 1. pp. 241-54.

RIS

TY - JOUR

T1 - Biosynthetic preparation of selectively deuterated phosphatidylcholine in genetically modified Escherichia coli

AU - Maric, Selma

AU - Thygesen, Mikkel B

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 - Pomorski, Thomas Günther

PY - 2015/1

Y1 - 2015/1

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 - Culture Media/chemistry

KW - Deuterium/metabolism

KW - Escherichia coli/genetics

KW - Metabolic Engineering

KW - Metabolic Networks and Pathways/genetics

KW - Phosphatidylcholines/metabolism

KW - Staining and Labeling/methods

U2 - 10.1007/s00253-014-6082-z

DO - 10.1007/s00253-014-6082-z

M3 - Article

C2 - 25301578

VL - 99

SP - 241

EP - 254

JO - Applied Microbiology and Biotechnology

JF - Applied Microbiology and Biotechnology

SN - 1432-0614

IS - 1

ER -