One-step purification of bacterial lipid macroamphiphiles by hydrophobic interaction chromatography.
Bacterial lipid macroamphiphiles extracted with phenol/water can be purified in one step by hydrophobic interaction chromatography. Lipids and the major part of protein are separated from macroamphiphiles during phenol/water extraction. Coextracted nucleic acids, polysaccharides, and residual protein are effectively removed by column chromatography on octyl-Sepharose whereby macroamphiphiles are primarily adsorbed and later eluted with a buffered propanol gradient. The procedure is applicable to macroamphiphiles with various lipid structures as was demonstrated using the diacylglycerol-containing lipoglycan of Micrococcus luteus, the lipid A-containing lipopolysaccharide of Salmonella typhimurium, and the diglyceryl tetraether lipoglycans of Thermoplasma acidophilum and Thermoplasma volcanicum. On elution from octyl-Sepharose, separation into molecular species of different compositions was observed with the lipopolysaccharide of S. typhimurium and the lipoglycan of T. volcanicum. It was also shown that, after phenol/water extraction, membrane lipids are completely recoverable from the phenol layer, which makes it possible to isolate lipids along with macroamphiphiles from the same sample of bacteria.
Fischer
W
W
Institut für Biochemie, Medizinische Fakultät, Universität Erlangen-Nürnberg, Germany.
D002847:Chromatography, Agarose / Q000379:methods*; D004987:Ethers / Q000302:isolation & purification; D008070:Lipopolysaccharides / Q000302:isolation & purification*; D008837:Micrococcus / Q000032:analysis; D010636:Phenols; D012486:Salmonella typhimurium / Q000032:analysis; D012995:Solubility; D013822:Thermoplasma / Q000032:analysis; D014867:Water
D004987:Ethers; D008070:Lipopolysaccharides; D010636:Phenols; C064351:lipomannan; D014867:Water