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Response surface methodology (RSM) was used for the determination of optimum extraction parameters to reach maximum lipid extraction yield with yeast.
The results revealed that the maximum lipid extraction yield was 18.1% obtained after extraction 120 min, extraction temperature 60 °C and M/EA ratio was 2 1.
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Of the tested methods, homogenization or direct solvent extraction (DSE) pretreatment, chloroform/methanol binary solvent system, and ≤600 µm particle size extracted maximum lipid of 22.1% irrespective of different biomass drying methods.
For this study, we defined the optimal time of MTBE-based lipid extraction at which maximum lipid recovery is achieved.
To demonstrate the variability of lipid extraction with varying conditions, we extracted the lipids from one biomass sample (Nannochloropsis sp).
Tissue lipids were extracted with Bligh-Dyer lipid extraction (Bligh and Dyer, 1959) and measured with the same enzymatic kits.
Lipid extraction and quantification of Q6 in yeast cell lipid extracts were determined by HPLC/MS MS as described [6,10].
Lipid extraction using hexane:diethyl ether (1 2 ratio) resulted in maximum extraction of lipids.
The results shown in Figure 4 (and Movie S3 of the Supporting Information) are essentially the same as those shown in Figure 2 (compare with the data for 1 μM daptomycin); in both cases, Δ A/ A decreased from a maximum by more than 4% that could be due to only lipid extraction.
At the time of lipid extraction, samples of bacterial pellets were removed and extracted using the same phospholipid extraction procedure.
Lipids were extracted from the cell lysates by adding 1 mL lipid extraction buffer (chloroform:methanol:acetic acid = 50 50 1, v/v/v).
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