Cellular Fatty Acids

Cellular fatty acids are analyzed after conversion into fatty acid methyl esters (FAMEs) by saponification, methylation and extraction following the protocol of Sasser (1990). The fatty acid methyl esters mixtures are separated by gas chromatography and detected by a flame ionization detector. In subsequent analysis, fatty acids are identified by a GC-MS run, on an Agilent GC-MS 7000D system (Vieira et. al., 2021). Peaks were identified based on retention time and mass spectra. For comparison with the literature we can still provide the naming of fatty acids according to the MIDI databases.
Further derivatization procedures for structural elucidation of yet unidentified compounds can be performed upon request. The position of single double bounds will be confirmed by a derivatization to the corresponding dimethyl disulfide adduct (Moss and Lambert-Fair, 1989). Branched-chain fatty acid positions, cyclo-positions and multiple double bounds could be determined by derivatization to their 3-pyridylcarbinol (“picolinyl”) and/or 4,4-dimethyloxazoline (DMOX) derivatives (Harvey, 1982, Spitzer, 1997, Yu et al., 1988).

Required material: For analysis of cellular fatty acids 30 mg freeze dried cells are required or 300 mg wet biomass frozen on dry ice or suspended in isopropanol (send at ambient temperature). Alternatively, an actively growing culture (still in exponential growth phase upon arrival, without signs of sporulation or cell lysis) could be send. Subculturing (if required) will be charged separately. If subculturing is requested by the customer, detailed cultivation conditions (media, temperature, oxygen supply, cultivation duration) must be provided.

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