Biosynthesis of terpenoids: 4-Diphosphocytidyl-2C-methyl-D-erythritol synthase of Arabidopsis thaliana
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.
- Publication:
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Proceedings of the National Academy of Science
- Pub Date:
- June 2000
- DOI:
- 10.1073/pnas.97.12.6451
- Bibcode:
- 2000PNAS...97.6451R
- Keywords:
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- Biochemistry