Article Figures & Data
Figures
- Fig. 1.
Mevalonate and nonmevalonate pathways for IPP biosynthesis. Fosmidomycin is a potent and specific inhibitor of DXP reductoisomerase in the nonmevalonate pathway. The reactions leading to IPP from CDP-ME2P remain unknown. MVA, mevalonate; PMVA, phosphomevalonate; DPMVA, diphosphomevalonate; TPP, thiamine diphosphate.
- Fig. 2.
Organization of ORFs in the 6.7-kbSnaBI-SnaBI fragment. The arrows indicate the extents and directions of the ORFs found in the flanking regions of thehmgr gene of Streptomyces sp. strain CL190. The arrowheads indicate the direction of the lacZ promoter in pUC118. pUMV19ΔM, derived from pUMV19, lacks only orfD.
- Fig. 3.
13C-NMR spectra of ubiquinone from JM109 and the transformant harboring pUMV19. Ubiquinone was purified from JM109 (lower), and its transformant harboring pUMV19 was grown in LB medium containing 0.1% sodium [1-13C]acetate under induction with 0.1 mM IPTG. The intensities of two signals at 61 ppm in both13C-NMR spectra were aligned. These signals were utilized as standards for the evaluation of isotopic abundance.
- Fig. 4.
Phenotypes of JM109 and the transformants. All plates are shown after overnight incubation at 37°C. Minimum medium M9 used in this experiment was always supplemented with 20 μg of thiamine/ml to meet the thiamine requirement of E. coli JM109. JM109, JM109(pUMV19), and JM109(pUMV19ΔM) could not grow on the M9 plate containing 20 μg of fosmidomycin (FMM)/ml (lower left), but JM109(pUMV19) and JM109(pUMV19ΔM) could grow on the plate under induction with 0.1 mM IPTG (lower right). On the lower-right plate, the growth of JM109(pUMV19ΔM) was slower than that of JM109(pUMV19).
Tables
- Table 1.
Deduced functions of orfA to orfE
ORF No. of aaa encoded Protein with the highest similarity (% identity/overlap [no. of aa]) Accession no. Reference orfA 345 Mevalonate kinase of M. jannaschii(26/306) Q58487 3 orfB 350 Diphosphomevalonate decarboxylase of S. cerevisiae (34/319) P32377 39 orfC 374 Mevalonate kinase of Arabidopsis thaliana(22/363) P46086 25 orfD 363 Hypothetical protein of E. herbicola(33/334) Q01335 1 orfE 389 HMG-CoA synthase of A. thaliana(28/371) P54873 22 ↵a aa, amino acids.
