Production and Analysis of Perdeuterated Lipids from Pichia pastoris Cells.

Alexis de Ghellinck (1, 2) , Hubert Schaller (3) , Valérie Laux (1) , Michael Haertlein (1) , Michele Sferrazza (2) , Eric Maréchal (4) , Hanna Wacklin (5, 6) , Juliette Jouhet (4) , Giovanna Fragneto (1)

1 ILL
2 ULB - Université libre de Bruxelles
3 IBMP - Institut de Biologie Moléculaire des Plantes
4 LPCV - Laboratoire de physiologie cellulaire végétale
5 ESS - European Spallation Source
6 Department of Chemistry [Copenhagen]

Probing molecules using perdeuteration (i.e deuteration in which all hydrogen atoms are replaced by deuterium) is extremely useful in a wide range of biophysical techniques. In the case of lipids, the synthesis of the biologically relevant unsaturated perdeuterated lipids is challenging and not usually pursued. In this work, perdeuterated phospholipids and sterols from the yeast Pichia pastoris grown in deuterated medium are extracted and analyzed as derivatives by gas chromatography and mass spectrometry respectively. When yeast cells are grown in a deuterated environment, the phospholipid homeostasis is maintained but the fatty acid unsaturation level is modified while the ergosterol synthesis is not affected by the deuterated culture medium. Our results confirm that the production of well defined natural unsaturated perdeuterated lipids is possible and gives also new insights about the process of desaturase enzymes.

Mots clés: deuterated medium; sterols; phospholipids; lipids; perdeuteration; yeast; Pichia pastoris; molecules probing; technique

Domaines: Biotechnologies; Chimie analytique

Alexis de Ghellinck, Hubert Schaller, Valérie Laux, Michael Haertlein, Michele Sferrazza, et al.. Production and Analysis of Perdeuterated Lipids from Pichia pastoris Cells.. PLoS ONE, 2014, 9 (4), pp.e92999. ⟨10.1371/journal.pone.0092999⟩. ⟨hal-00985220⟩