Triglyceride synthesis involves acyl transfer between long chain acyl coenzyme A molecules and the hydroxyl groups of glycerol 3-phosphate (formed by reduction of dihydroxyacetone 3-phosphate from glycolysis). The phosphatidic acid formed (two fatty acid chains and a phosphate group, phosphatidate) can be followed by hydrolysis of the phosphate ester and an additional acyl exchange (adding the third fatty acid chain) which leads to a triglyceride. Alternatively, the phosphatidic acid can serve as the starting point for membrane phospholipid synthesis along one of several routes.

One pathway of phosphoglyceride synthesis begins with the activation of phosphatidate with CTP to form CDP diacylglycerol, a process thermodynamically driven by pyrophosphate cleavage. The molecule is then activated for phosphate esterification with an alcohol such as serine, to form phosphatidyl serine or with an alcohol such as inositol to form inositol 4,5-bisphosphate, a key molecule in signal transduction. Phosphatidyl serine can then serve in bacteria as a precursor to phosphatidyl choline, a key component of cell membranes. In mammals, choline from the diet is used to form CDP-choline which substitutes directly onto diacylglycerol to form phosphatidyl choline.