Biosynthesis of terpenoids: 4-diphosphocytidyl-2-C-methyl-D-erythritol kinase from tomato

The putative catalytic domain (residues 81-401) of a predicted tomato prote in with similarity to 4-diphosphocytidyl-2-C-methyl-D-erythritol kinase of Escherichia coli was expressed in a recombinant E. coli strain. The protein was purified to homogeneity and was shown to catalyze the phosphorylation of the position 2 hydroxy group of 4-diphosphocytidyl-2-C-methyl-D-erythrit ol at a rate of 33 mu mol.mg(-1) min(-1). The structure of the reaction pro duct, 4-diphosphocytidyl-2-C-methyl-D-erythrit 2-phosphate, was established by NMR spectroscopy. Divalent metal ions, preferably Mg2+, are required fo r activity. Neither the tomato enzyme nor the E. coli ortholog catalyzes th e phosphorylation of isopentenyl monophosphate.