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.
MeSH terms
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Cell Membrane / metabolism
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Cell Proliferation
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Culture Media / chemistry
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Deuterium / chemistry
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Deuterium / metabolism*
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Fatty Acids / biosynthesis
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Fatty Acids / metabolism
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Phospholipids / biosynthesis*
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Phospholipids / chemistry
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Pichia / cytology*
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Pichia / metabolism*
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Sterols / biosynthesis*
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Sterols / chemistry
Substances
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Culture Media
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Fatty Acids
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Phospholipids
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Sterols
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Deuterium
Grants and funding
VL and MH acknowledge Engineering and Physical Science Research Council support for the Dlab to V.T. Forsyth, Keele University, under grant EP/C015452/1 and from the EU under contract RII3-CT-2003-505925. JJ and EM were supported by a Blue Sky grant from Agence Nationale de la Recherche (ANR ReGal, ANR- 10-BLAN-1524). This research project has been supported by the European Commission under the 7th Framework Programme through the “Research Infrastructures” action of the “Capacities” Programme, NMI3-II Grant number 283883. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.