2.7 Extraction and work-up of fatty alcohols from bioreactor cultivation for acetylation reaction
The 2.4 L of broth from the bioreactor cultivation was centrifuged at 3,000 g for 5 min at room temperature, and the supernatant was separated from the pellet. Fatty alcohols were extracted from the supernatant by adding 2.2 L of ethyl acetate and incubating on a multi-vortexer for 1 h. Organic phase was decanted. To extract fatty alcohols from the pellet, 100 mL of ethyl acetate was added to the pellet, and the mix was shaken on a multi-vortexer for 6 h . The organic phase was decanted, 100 mL of fresh solvent was added to the biomass pellet, and the mixing repeated for 1 h. The organic phase was decanted. All ethyl acetate fractions from the supernatant and pellet extraction were combined and dried with anhydrous sodium sulfate. The solvent was removed in a rotary evaporator.
The extracted crude fatty alcohol mixture was passed through a silica gel column, which was prepared as described in [20]. The silica was washed with hexane and then subsequently with a gradient of hexane/ethyl acetate at the proportion of 95:5 to 40:60 (% v/v). Purest fractions based on thin-layer chromatography were collected, and the solvent was evaporated in a rotary evaporator.