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.