Seven solvent mixtures have been used to extract the lipid fraction of lyophilized biomass ofIsochrysis galbana. Six of them were composed of biocompatible solvents. Each method was carried out under relaxed operating conditions (i.e., one hour at room temperature) with extraction in a nitrogen atmosphere to prevent autooxidation and degradation of polyunsaturated fatty acids (PUFAs). Apart from the well‐established Bligh and Dyer method [Can. J. Biochem. Physiol. 37:911 (1959)] (Cl3CH/MeOH/H2O, 1∶2∶0.8, vol/vol/vol), which rendered the highest yield of lipids (93.8%), ethanol (96%) and hexane/ethanol (96%), 1∶2.5 vol/vol produced the best results (84.4 and 79.6%, respectively). To obtain free fatty acids, KOH was added to the solvent mixtures used to extract the total lipids, except for Cl3CH/MeOH/H2O, and direct saponification was carried out at 60°C for 1 h or at room temperature for 8 h. The highest yields obtained by direct saponicification were 81% with hexane/ethanol (96%), 1∶2.5, vol/vol and 79.8% with ethanol (96%). Partial yields of the mainn‐3 PUFAs found inI. galbana, stearidonic acid (SA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), were calculated for both extraction methods. For lipid extraction with ethanol (96%), yields of 91, 82 and 83% were obtained for SA, EPA and DHA, respectively. When direct saponification was used, hexane/ethanol (96%; 1∶2.5, vol/vol) produced the best yields of (91, 79 and 69% for SA, EPA and DHA, respectively).
for the preparation of fatty acid methyl esters allows notable saving of time and reagents. The material being analysed is heated for 10 minutes with methanol, acetyl chloride and hexane.
A 20 L gas-agitated slurry bubble column bioreactor was used to investigate the effects of nonmechanical low-intensity agitation on development of broth rheology and fungal pellet morphology during production of lovastatin by the filamentous fungus Aspergillus terreus. Fermentations were carried out under elevated dissolved oxygen levels (400% of air saturation) at gassing rates that ranged from 0.5 to 1.5 vvm. Various initial concentrations of the growth limiting nitrogen source were used to attain different total biomass concentrations, to observe the effect of this variable on development of pellets and the rheology of the fermentation broth. The non-Newtonian rheology of the fermentation broth was influenced both by the biomass concentration and the size of the fungal pellets. The stable pellet diameter ranged from ∼2300 to ∼2900 µm. Too low turbulence (gassing rate of 0.5 vvm) and low dissolved oxygen levels adversely affected lovastatin production. The best biomass specific production of lovastatin was attained at high biomass concentrations under oxygen-rich conditions that were not excessively turbulent. In fermentation broths with various rheologies, the oxygen-transfer coefficient in the bubble column correlated with the aeration velocity, biomass concentration, and effective viscosity of the broth. The correlations obtained were significantly different for broths with pelleted growth and those with filamentous growth.
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