Methyl non adecaanoate c19

Fatty acid methyl esters (FAMEs) were obtained by methanol transesterification of lyophilized yeast biomass. The transesterification protocol was originally adopted from Griffiths et al. (2010 (link)) and modified in our lab by Gorner et al. (2016 (link)). FAME profiles were analyzed on a GC-2010 Plus gas chromatograph from Shimadzu (Nakagyo-ku, Kyoto, Japan) with flame ionization detector. One microliter sample was applied by AOC-20i auto injector (Shimadzu) onto a ZB-WAX column [30 m, 0.32 mm ID; 0.25 μm df; phenomenex (Torrance, CA, USA)]. The initial column temperature was set at 150°C (maintained for 1 min). A temperature gradient was applied from 150° to 240°C (5°C.min⁻¹), followed by 6 min maintenance at 240°C. Fatty acids were identified according to retention times of the authentic standard: Marine Oil FAME Mix (Restek, USA). Individual FAME concentrations were based on peak areas relative to Methyl Non-adecaanoate C19 (Sigma, Germany), which was incorporated as an internal standard in all samples.

The extent of lipid contamination in protein pellets, following various purification methods, was measured by accounting for the sum of fatty acid methyl esters (FAMEs), obtained by methanol transesterification. The transesterification protocol was originally adopted from [46 (link)]. and modified in our lab by [47 (link)]. FAME profiles were analyzed on a GC-2025 gas chromatograph from Shimadzu (Nakagyo-ku, Kyōto, Japan) with flame ionization detector. One microliter sample was applied by AOC-20i auto injector (Shimadzu) onto a ZB-WAX column (30 m, 0.32 mm ID; 0.25 μm df; phenomenex (Torrance, CA, USA)). The initial column temperature was 150 °C (maintained for 1 min). A temperature gradient was applied from 150–240 °C (5 °C min⁻¹), followed by 6 min maintenance at 240 °C. Fatty acids were identified according to retention times of the authentic standard: Marine Oil FAME Mix (Restek, USA). Individual FAME concentrations were based on peak areas relative to Methyl Nonadecaanoate C19 (Sigma, Germany), which was incorporated as an internal standard in all samples. Percent lipid was calculated from the sum of individually identified FAMEs with respect to pellet dry weight.