Phosphorylation of 1-deoxy-D-xylulose by D-xylulokinase of Escherichia coli

Wungsintaweekul, Juraithip; Herz, Stefan; Hecht, Stefan; Eisenreich, Wolfgang; Feicht, Richard; Rohdich, Felix; Bacher, Adelbert; Zenk, Meinhart H.

doi:10.1046/j.1432-1327.2001.01875.x

Cited by 25 publications

(35 citation statements)

References 18 publications

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“…Importantly, the inhibitory effect of ketoclomazone was suppressed by the addition of 0.1% DX to the growth medium. As DX has been reported to be converted into DXP by the action of D-xylulokinase and incorporated into the MEP pathway for IPP biosynthesis in E. coli (Figure 1), 32 DX should be incorporated into the MEP pathway in H. influenzae as well. Therefore, the suppression caused by the addition of DX strongly suggested that ketoclomazone inhibited DXP synthases of E. coli and H. influenzae.…”

Section: Dxp Synthase Inhibition Assaysmentioning

confidence: 99%

The herbicide ketoclomazone inhibits 1-deoxy-D-xylulose 5-phosphate synthase in the 2-C-methyl-D-erythritol 4-phosphate pathway and shows antibacterial activity against Haemophilus influenzae

et al. 2010

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Two distinct metabolic pathways have been elucidated for the formation of isopentenyl diphosphate and dimethylallyl diphosphate, essential metabolic precursors for isoprenoid biosynthesis: the mevalonate pathway, found ubiquitously in mammals, and the 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway, found in most bacteria. As the MEP pathway is absent from mammals, all MEP pathway enzymes represent effective targets for the development of antibacterial drugs. In this study, we found that a herbicide, ketoclomazone, exhibited antibacterial activity against a pathogenic bacterium, Haemophilus influenzae, with an MIC value of 12.5 lg ml À1 and that antibacterial activity was suppressed by adding 1-deoxyxylulose, a free alcohol of 1-deoxy-D-xylulose 5-phosphate (DXP). DXP is an MEP pathway intermediate synthesized from pyruvate and D-glyceraldehyde 3-phosphate (D-GAP) by the action of DXP synthase. Thus, we investigated the enzyme kinetics of DXP synthase of H. influenzae (HiDXS) to elucidate an inhibitory mechanism of ketoclomazone on HiDXS. The dxs gene was cloned from H. influenzae and overexpressed in Escherichia coli, and the enzyme was purified to homogeneity. The purified HiDXS was a soluble dimeric 70-kDa protein. Steady-state kinetic constants for HiDXS were calculated, and Lineweaver-Burk plots were consistent with a ping-pong bi bi mechanism. The kinetics of inhibition by ketoclomazone suggested that ketoclomazone binds to an unidentified inhibitor-binding site that differs from both the pyruvate-binding site and the D-GAP-binding site on DXP synthase. These data reveal the inhibitory mechanism of ketoclomazone on DXP synthase.

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Section: Dxp Synthase Inhibition Assaysmentioning

confidence: 99%

The herbicide ketoclomazone inhibits 1-deoxy-D-xylulose 5-phosphate synthase in the 2-C-methyl-D-erythritol 4-phosphate pathway and shows antibacterial activity against Haemophilus influenzae

et al. 2010

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“…1). In silico studies have indicated the presence of a putative DOX kinase in B. subtilis with high sequence similarity to an E. coli enzyme, 27) suggesting that B. subtilis has the capability to convert DOX to DOXP in vivo, potentially increasing flux through the MEP pathway. As seen in Fig.…”

Section: Addition Of Carbon Skeletons To the Mep Pathway Of B Subtilismentioning

confidence: 99%

Evidence of Isoprenoid Precursor Toxicity inBacillus subtilis

Sivy

Fall

Rosenstiel

2011

Bioscience, Biotechnology, and Biochemistry

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“…Therefore, Dxs-produced 1-deoxy-D-xylulose-5-phosphate is an unlikely intermediate in the PdxST-dependent pathway of PLP synthesis. Though an alternative route of 1-deoxy-D-xylulose-5-phosphate synthesis was described in E. coli, its physiological role is unknown (43).…”

Section: Figmentioning

confidence: 99%

Physical and Enzymological Interaction of Bacillus subtilis Proteins Required for De Novo Pyridoxal 5′-Phosphate Biosynthesis

Belitsky

2004

J Bacteriol

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Bacillus subtilis synthesizes pyridoxal 5-phosphate, the active form of vitamin B 6 , by a poorly characterized pathway involving the yaaD and yaaE genes. The pdxS (yaaD) mutant was confirmed to be a strict B 6 auxotroph, but the pdxT (yaaE) mutant turned out to be a conditional auxotroph depending on the availability of ammonium in the growth medium. The PdxS and PdxT proteins copurified during affinity chromatography and apparently form a complex that has glutaminase activity. PdxS and PdxT appear to encode the synthase and glutaminase subunits, respectively, of a glutamine amidotransferase of as-yet-unknown specificity essential for B 6 biosynthesis.

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Phosphorylation of 1-deoxy-D-xylulose by D-xylulokinase of Escherichia coli

Cited by 25 publications

References 18 publications

The herbicide ketoclomazone inhibits 1-deoxy-D-xylulose 5-phosphate synthase in the 2-C-methyl-D-erythritol 4-phosphate pathway and shows antibacterial activity against Haemophilus influenzae

The herbicide ketoclomazone inhibits 1-deoxy-D-xylulose 5-phosphate synthase in the 2-C-methyl-D-erythritol 4-phosphate pathway and shows antibacterial activity against Haemophilus influenzae

Evidence of Isoprenoid Precursor Toxicity inBacillus subtilis

Physical and Enzymological Interaction of Bacillus subtilis Proteins Required for De Novo Pyridoxal 5′-Phosphate Biosynthesis

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