期刊
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
卷 99, 期 1, 页码 241-254出版社
SPRINGER
DOI: 10.1007/s00253-014-6082-z
关键词
Glycerophospholipids; Phosphatidylcholine; Selective deuteration; Biosynthesis; E. coli; Neutron scattering; NMR; Mass spectrometry; Evolution in the test tube
资金
- European Spallation Source
- UNIK research initiative of the Danish Ministry of Science, Technology and Innovation through the 'Center for Synthetic Biology' at University of Copenhagen
- Danish National, and the Research Foundation through the PUMPKIN Center of Excellence
- German Research Council [SFB 1052/B6]
- UK Engineering and Physical Sciences Research Council (EPSRC) [GR/R99393/01, EP/C015452/1]
- EU NMI3 programme
- EU CRISP programme [283745]
- United States National Institutes of Health [GM R37-20748]
- John Dunn Research Foundation
- EPSRC [EP/C015452/1] Funding Source: UKRI
- Engineering and Physical Sciences Research Council [GR/R99393/01, EP/C015452/1] Funding Source: researchfish
- NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES [R37GM020478] Funding Source: NIH RePORTER
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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