Biosynthesis of terpenoids: 4-Diphosphocytidyl-2C-methyl- d -erythritol synthase of Arabidopsis thaliana

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Abstract
A hypothetical gene with similarity to the ispD gene of Escherichia coli was cloned from Arabidopsis thaliana cDNA. The ORF of 909 bp specifies a protein of 302 amino acid residues. The cognate chromosomal gene consists of 2,071 bp and comprises 11 introns with a size range of 78–202 bp. A fragment comprising amino acid residues 76–302 was expressed in a recombinant E. coli strain. The protein was purified to homogeneity and was shown to catalyze the formation of 4-diphosphocytidyl-2C-methyl-d-erythritol from 2C-methyl-d-erythritol 4-phosphate with a specific activity of 67 μmol⋅min−1 mg−1. The Michaelis constants for 4-diphosphocytidyl-2C-methyl-d-erythritol and CTP were 500 μM and 114 μM, respectively.