In this study a pathway for the synthesis of triacylglycerol (TAG) within the lumen of the endoplasmic reticulum has been identified, using microsomes that had been preconditioned by depleting their endogenous substrates and then fusing them with biotinylated phosphatidylserine liposomes containing CoASH and Mg
Hydrolysis of groundnut oil and synthetic glyceryl tri(~leate-l-~~C) by bovine rumen contents was demonstrated under in vitro conditions. Fractionation by differential centrifugation of total rumen contents obtained from a cow grazing on fresh pasture showed that the organisms responsible for hydrolysis of triglycerides were closely associated with the particulate material in the rumen and homogenisation released much of the lipolytic activity. Cell-free extracts of mixed rumen bacteria were prepared by subjecting mixed rumen bacteria to high frequency sonication or osmotic shock followed by isolation of the released lipolytic enzymes by high-speed centrifugation.A method for assaying lipase activity is described which uses glyceryl tri(oleate-l-l*C) as substrate, and thin-layer chromatography to separate the free fatty acids, mono-, di-and tri-glycerides prior to radiochemical determination. The method was developed using pancreatic lipase and was applied to the isolation of lipases from rumen micro-organisms.
IntroductionAlthough early investigations on lipid metabolism in the rumen established the importance of biohydrogenation in changing the composition of the dietary lipids,' it has only recently been realised that no hydrogenation of unsaturated fatty acids takes place unless the constituent fatty acids of triglycerides are first released in free form by the activity of the lipolytic micro-organisms.2 The relationship between the lipolysis and biohydrogenation of other dietary constituents has not been studied in such detail but from a comparison of the fatty acid composition of neutral and polar lipids and of free fatty acids in the rumen and in the diet it would seem
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