[53] Preparation of phospholipids and their analogs by phospholipase D

摘要: Publisher Summary This chapter discusses the preparation of phospholipids and their analogs by phospholipase D. Egg phosphatidylcholine was extracted from egg yolk purified on silica gel 60, using solvent mixtures chloroform-methanol-ammonia in ratios 200:15:1 to start with 65: 15:1 for elution phosphatidylcholine. The pure lipid dissolved chloroform-methanol, 9:1 (by volume), stored at 0° under nitrogen. transfer phosphatidyl residue primary alcohols or ethanolamines detected TLC system 65:30:3. hydrolysis phosphatidylethanolamines performed presence Triton X-100, which also facilitated formation clear emulsions thus allowed complete substrates. It is found that no difference extent rate transphosphatidylation observed when smaller samples were used experiments respectively reduced amounts enzyme alkanol volumes buffer–ether mixture.