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 action of IspD, IspE and IspF proteins in the non-mevalonate pathway of isoprenoid biosynthesis. We describe here rapid one-pot strategies for the enzyme-assisted preparation of this compound from isotope-labeled pyruvate and glucose that are optimized for the introduction of 13C or 14C and afford a wide variety of isotopomers in mmol quantity. The one-pot reactions involving up to 10 forward reaction steps and up to 15 enzyme catalysts have an overall yield of 50 to 80% of purified product.