Biosynthesis of terpenoids

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Abstract
The putative catalytic domain of an open reading frame from Plasmodium falciparum with similarity to the ispF gene of Escherichia coli specifying 2C-methyl-d-erythritol 2,4-cyclodiphosphate synthase was expressed in a recombinant E. coli strain. The recombinant protein was purified to homogeneity and was found to catalyze the formation of 2C-methyl-d-erythritol 2,4-cyclodiphosphate from 4-diphosphocytidyl-2C-methyl-d-erythritol 2-phosphate at a rate of 4.3 µmol·mg−1·min−1. At lower rates, the recombinant protein catalyzes the formation of 2-phospho-2C-methyl-d-erythritol 3,4-cyclophosphate from 4-diphosphocytidyl-2C-methyl-d-erythritol 2-phosphate and the formation of 2C-methyl-d-erythritol 3,4-cyclophosphate from 4-diphosphocytidyl-2C-methyl-d-erythritol. Divalent metal ions such as magnesium or manganese are required for catalytic activity. The enzyme has a pH optimum at pH 7.0. Recombinant expression of the full-length open reading frame afforded insoluble protein that could not be folded in vitro. The enzyme is a potential target for antimalarial drugs directed at the nonmevalonate pathway of isoprenoid biosynthesis.