The extraction of inositol-containing phospholipids and phosphatidylcholine from Saccharomyces cerevisiae and Neurospora crassa.
By use of fungi grown in the presence of [3H]-inositol and [14C]choline, we have explored methods for the quantitative extraction of inositol-containing phospholipids and phosphatidylcholine. Slightly alkaline mixtures of both ethanol-water and ethanol-diethylether-water at elevated temperatures were shown to effectively extract these lipids from intact Saccharomyces cerevisiae and Neurospora crassa. Some previously published procedures fail to completely extract to very polar phosphoinositol-containing sphingolipids of these organisms. Trichloroacetic acid can be used with caution in killing cells prior to extraction; lipid destruction can occur at elevated concentrations and temperatures. Complete extraction of these very polar lipids with polar solvents also results in an extract containing significant amounts of non-lipids.
D007294:Inositol / Q000302:isolation & purification*; D008722:Methods; D009491:Neurospora / Q000032:analysis*; D009492:Neurospora crassa / Q000032:analysis*; D010713:Phosphatidylcholines / Q000302:isolation & purification*; D010743:Phospholipids / Q000302:isolation & purification*; D012441:Saccharomyces cerevisiae / Q000032:analysis*; D012997:Solvents; D014238:Trichloroacetic Acid
D010713:Phosphatidylcholines; D010743:Phospholipids; D012997:Solvents; D007294:Inositol; D014238:Trichloroacetic Acid