title: Biodiesel and poly-unsaturated fatty acids production from algae and crop plants - a rapid and comprehensive workflow for lipid analysis.
abstract: Fatty acid methyl ester analysis (FAME) by gas chromatography coupled to mass spectrometry (GC-MS) is a widely used technique in biodiesel/bioproduct (e.g. poly-unsaturated fatty acids, PUFA) research but typically does not allow distinguishing between bound and free fatty acids. To understand and optimize biosynthetic pathways, however, the origin of the fatty acid is an important information. Furthermore the annotation of PUFAs is compromised in classical GC-EI-MS because the precursor molecular ion is missing. In the present protocol an alkaline methyl esterification step with TMS derivatization enabling the simultaneous analysis of bound and free fatty acids but also further lipids such as sterols in one GC-MS chromatogram is combined. This protocol is applied to different lipid extracts from single cell algae to higher plants: Chlorella vulgaris, Chlamydomonas reinhardtii, Coffea arabica, Pisum sativum and Cuscuta japonica. Further, field ionization (GC-FI-MS) is introduced for a better annotation of fatty acids and exact determination of the number of double bonds in PUFAs. The proposed workflow provides a convenient strategy to analyze algae and other plant crop systems with respect to their capacity for third generation biodiesel and high-quality bioproducts for nutrition such as PUFAs.
authors.affiliation: Department of Ecogenomics and Systems Biology, Vienna University, Vienna, Austria. takeshi.furuhashi@univie.ac.at.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Furuhashi
authors.affiliation: Collaboration Promotion Unit, RIKEN Global Research Cluster, Wako,, Saitama, Japan.
authors.forename: Takemichi
authors.initials: T
authors.lastname: Nakamura
authors.affiliation: Department of Ecogenomics and Systems Biology, Vienna University, Vienna, Austria.
authors.forename: Lena
authors.initials: L
authors.lastname: Fragner
authors.affiliation: Department of Ecogenomics and Systems Biology, Vienna University, Vienna, Austria.
authors.forename: Valentin
authors.initials: V
authors.lastname: Roustan
authors.affiliation: Department of Ecogenomics and Systems Biology, Vienna University, Vienna, Austria.|Ecoduna Produktions GmbH, Bruck/Leitha, Austria.
authors.forename: Verena
authors.initials: V
authors.lastname: Schön
authors.affiliation: Department of Ecogenomics and Systems Biology, Vienna University, Vienna, Austria. wolfram.weckwerth@univie.ac.at.|Vienna Metabolomics Center (VIME), Vienna University, Vienna, Austria. wolfram.weckwerth@univie.ac.at.
authors.forename: Wolfram
authors.initials: W
authors.lastname: Weckwerth
mesh_terms: D056804:Biofuels / Q000032:analysis*; D053898:Biosynthetic Pathways; D016825:Chlamydomonas reinhardtii / Q000378:metabolism; D048408:Chlorella vulgaris / Q000378:metabolism; D000460:Chlorophyta / Q000378:metabolism*; D040503:Coffea / Q000378:metabolism; D034182:Cuscuta / Q000378:metabolism; D005231:Fatty Acids, Unsaturated / Q000032:analysis* / Q000378:metabolism; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D019684:Magnoliopsida / Q000378:metabolism*; D018532:Pisum sativum / Q000378:metabolism; D059010:Single-Cell Analysis
chemical_list: D056804:Biofuels; D005231:Fatty Acids, Unsaturated
keywords: Biofuels; Biomass; Lipidomics; PUFA; high value nutrition