Abstract
A procedure has been developed for the separation of the glycosyl diglycerides from the phosphatides of Gram-positive bacteria on columns of silicic acid. The method utilizes mixtures of acetone in chloroform for elution of the glycosyl diglycerides, followed by increasing amounts of methanol in chloroform for elution of the phosphatides. The course of the fractionation was followed by means of phosphorus and carbohydrate determinations and by paper chromatography. The completeness of the separation of the phosphatides from the sugar-containing lipids was shown also by chromatographing a total lipid extract containing P32-labeled phosphatides.
Highlights
Growth of CellsThe organisms used in this study were Streptococcusfaecalis (ATCC 9790) and Lactobacillus plantarum B-246
Similar observations have been made with plant lipids, which are complicated by the presence of glycosyl diglycerides [6,7,8,9]
The glycosyl diglycerides were fractionated by use of increasing increments of acetone in chloroform, and the phosphatides by use of increasing amounts of methanol in chloroform
Summary
The organisms used in this study were Streptococcusfaecalis (ATCC 9790) and Lactobacillus plantarum B-246. Six-liter Florence flasks containing 4.5 liters of medium were inoculated with 45 ml of an 18 hr culture grown in the same medium. S.faecalis was grown at 37'; L.plantarum a t 32'. 7H20,lOO g; FeSOl. 7 H z 0 , 5 g; NaCl, 5 g; MnS04.7Hz0, 2 g, dissolved in 500 ml distilled water. The cell paste was washed three times with cold distilled water.
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