Studies on the non-mevalonate pathway - Preparation and properties of isotope-labeled 2C-methyl-D-erythritol 2,4-cyclodiphosphate

Recent studies have shown that 2C-methyl-D-erythritol 2,4-cyclodiphosphate can be formed from 2C-methyl-D-erythritol 4-phosphate by the consecutive ac tion of IspD, IspE and IspF proteins in the non-mevalonate pathway of isopr enoid biosynthesis. We describe here rapid one-pot strategies for the enzym e-assisted preparation of this compound from isotope-labeled pyruvate and g lucose that are optimized for the introduction of C-13 or C-14 and afford a wide variety of isotopomers in mmol quantity. The one-pot reactions involv ing up to 10 forward reaction steps and up to 15 enzyme catalysts have an o verall yield of 50 to 80% of purified product.