Biodiesel and poly-unsaturated fatty acids production from algae and crop plants - a rapid and comprehensive workflow for lipid analysis

Author(s)
Takeshi Furuhashi, Takemichi Nakamura, Lena Fragner, Valentin Roustan, Verena Schoen, Wolfram Weckwerth
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.

Organisation(s)
Department of Ecogenomics and Systems Biology, Large-Instrument Facility for Mass Spectrometry in Life Sciences, Research Platform Vienna Metabolomics Center
External organisation(s)
RIKEN Advanced Science Institute, Ecoduna Prod GmbH
Journal
Biotechnology Journal
Volume
11
Pages
1262-1267
No. of pages
6
ISSN
1860-6768
DOI
https://doi.org/10.1002/biot.201400197
Publication date
10-2016
Peer reviewed
Yes
Austrian Fields of Science 2012
Spectroscopy
Keywords
Portal url
https://ucris.univie.ac.at/portal/en/publications/biodiesel-and-polyunsaturated-fatty-acids-production-from-algae-and-crop-plants--a-rapid-and-comprehensive-workflow-for-lipid-analysis(0968432d-cc50-42b4-8ebb-a088f70b1688).html