title: Production and analysis of perdeuterated lipids from Pichia pastoris cells.
abstract: 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.
authors.affiliation: Institut Laue-Langevin, Grenoble, France; Service des polymères, Université Libre de Bruxelles, Brussels, Belgium.
authors.forename: Alexis
authors.initials: A
authors.lastname: de Ghellinck
authors.affiliation: Institut de Biologie Moléculaire des Plantes du CNRS, Strasbourg, France.
authors.forename: Hubert
authors.initials: H
authors.lastname: Schaller
authors.affiliation: Institut Laue-Langevin, Grenoble, France.
authors.forename: Valérie
authors.initials: V
authors.lastname: Laux
authors.affiliation: Institut Laue-Langevin, Grenoble, France.
authors.forename: Michael
authors.initials: M
authors.lastname: Haertlein
authors.affiliation: Service des polymères, Université Libre de Bruxelles, Brussels, Belgium.
authors.forename: Michele
authors.initials: M
authors.lastname: Sferrazza
authors.affiliation: Laboratoire de physiologie cellulaire et végétale, CNRS/CEA/Univ. Grenoble Alpes/INRA, Grenoble, France.
authors.forename: Eric
authors.initials: E
authors.lastname: Maréchal
authors.affiliation: European Spallation Source ESS AB, Lund, Sweden; Chemistry Department, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Hanna
authors.initials: H
authors.lastname: Wacklin
authors.affiliation: Laboratoire de physiologie cellulaire et végétale, CNRS/CEA/Univ. Grenoble Alpes/INRA, Grenoble, France.
authors.forename: Juliette
authors.initials: J
authors.lastname: Jouhet
authors.affiliation: Institut Laue-Langevin, Grenoble, France.
authors.forename: Giovanna
authors.initials: G
authors.lastname: Fragneto
mesh_terms: D002462:Cell Membrane / Q000378:metabolism; D049109:Cell Proliferation; D003470:Culture Media / Q000737:chemistry; D003903:Deuterium / Q000737:chemistry / Q000378:metabolism*; D005227:Fatty Acids / Q000096:biosynthesis / Q000378:metabolism; D010743:Phospholipids / Q000096:biosynthesis* / Q000737:chemistry; D010843:Pichia / Q000166:cytology* / Q000378:metabolism*; D013261:Sterols / Q000096:biosynthesis* / Q000737:chemistry
chemical_list: D003470:Culture Media; D005227:Fatty Acids; D010743:Phospholipids; D013261:Sterols; D003903:Deuterium
references.citation: Plant Cell Physiol. 2006 Jan;47(1):64-73
references.pmid: 16267098
references.citation: Acta Crystallogr D Biol Crystallogr. 1997 Jul 1;53(Pt 4):377-84
references.pmid: 15299903
references.citation: Biochim Biophys Acta. 2007 Mar;1771(3):271-85
references.pmid: 16920014
references.citation: Biochim Biophys Acta. 2007 Apr;1771(4):455-61
references.pmid: 17293161
references.citation: Yeast. 2001 Jun;18(8):679-95
references.pmid: 11378896
references.citation: J Bacteriol. 1989 Dec;171(12):6409-13
references.pmid: 2687232
references.citation: Genetics. 2012 Feb;190(2):317-49
references.pmid: 22345606
references.citation: J Biol Chem. 1997 Aug 15;272(33):20873-83
references.pmid: 9252414
references.citation: Plant Mol Biol. 2004 Feb;54(3):335-52
references.pmid: 15284491
references.citation: Biochim Biophys Acta. 2000 Nov 10;1469(3):159-95
references.pmid: 11063882
references.citation: Biophys J. 1998 Dec;75(6):3057-63
references.pmid: 9826625
references.citation: FEBS Lett. 1979 Jun 15;102(2):244-8
references.pmid: 156647
references.citation: Biochim Biophys Acta. 2009 Mar;1791(3):166-72
references.pmid: 19168151
references.citation: Biochim Biophys Acta. 1981 Sep 24;665(3):399-407
references.pmid: 6457645
references.citation: Nat Prod Rep. 2004 Apr;21(2):249-62
references.pmid: 15042148
references.citation: J Biomol NMR. 2000 Aug;17(4):337-47
references.pmid: 11014598
references.citation: Appl Microbiol Biotechnol. 2010 Jul;87(4):1347-54
references.pmid: 20405122
references.citation: Proc Natl Acad Sci U S A. 1975 Jan;72(1):376-80
references.pmid: 16592215
references.citation: J Mol Biol. 1979 Nov 15;134(4):693-706
references.pmid: 537075
references.citation: Eur J Biochem. 1986 Nov 17;161(1):191-6
references.pmid: 3780736
references.citation: Biochemistry. 2003 Mar 18;42(10):3054-9
references.pmid: 12627972
references.citation: J Biol Chem. 1957 May;226(1):497-509
references.pmid: 13428781
references.citation: J Mol Biol. 1985 Dec 5;186(3):565-82
references.pmid: 4093979
references.citation: Appl Microbiol Biotechnol. 2012 Jan;93(1):31-9
references.pmid: 22057543
references.citation: Biophys Chem. 2000 Aug 30;86(2-3):249-57
references.pmid: 11026689
references.citation: Science. 2000 Jun 2;288(5471):1604-7
references.pmid: 10834833
references.citation: Yeast. 2005 Mar;22(4):249-70
references.pmid: 15704221
references.citation: Biochim Biophys Acta. 2013 Mar;1831(3):523-32
references.pmid: 23010477
references.citation: J Biomol NMR. 1999 Jul;14(3):231-9
references.pmid: 10481275
references.citation: PLoS One. 2013;8(2):e56429
references.pmid: 23409184
references.citation: Mol Biol Cell. 2012 Apr;23(7):1146-56
references.pmid: 22323296
references.citation: Plant Mol Biol. 2001 Jan;45(1):75-92
references.pmid: 11247608
references.citation: J Biol Chem. 2007 Dec 28;282(52):37293-7
references.pmid: 17981800
references.citation: Biochem Soc Trans. 2002 Nov;30(Pt 6):1080-2
references.pmid: 12440977
references.citation: Eur Biophys J. 2007 Apr;36(4-5):281-91
references.pmid: 17333162
references.citation: Proc Natl Acad Sci U S A. 2005 Aug 16;102(33):11639-44
references.pmid: 16087874
references.citation: Biochim Biophys Acta. 1975 Mar 24;380(3):442-53
references.pmid: 166662