Biosynthesis of , a putative C5 intermediate in the mevalonate independent pathway for isoprenoid biosynthesis

Duvold, Tore; Bravo, Jean-Michel; Pale-Grosdemange, Catherine; Rohmer, Michel

doi:10.1016/s0040-4039(97)01045-9

Cited by 92 publications

(59 citation statements)

References 18 publications

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“…As part of an ongoing effort to isolate and define genes involved in isoprenoid metabolism, we analyzed the sequences of 1,300 random clones obtained from an enriched cDNA library constructed specifically from mRNA isolated from peppermint glandular trichome secretory cells. Because the most advanced, defined intermediate of the plastidial DXP pathway to isoprenoids is MEP (19)(20)(21), and the end product of the pathway is IPP (25, 26), a phosphorylation step must occur at some point during this reaction sequence. Accordingly, metabolite kinases were sought, but only two clones with similarity to adenylate kinases were noted by searching the common databases.…”

Section: Cloning and Heterologous Expression Of A Putative Kinase Frommentioning

confidence: 99%

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Isopentenyl diphosphate biosynthesis via a mevalonate-independent pathway: Isopentenyl monophosphate kinase catalyzes the terminal enzymatic step

Croteau

1999

Proc. Natl. Acad. Sci. U.S.A.

103

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In plants, the biosynthesis of isopentenyl diphosphate, the central precursor of all isoprenoids, proceeds via two separate pathways. The cytosolic compartment harbors the mevalonate pathway, whereas the newly discovered deoxyxylulose 5-phosphate pathway, which also operates in certain eubacteria, including Escherichia coli, is localized to plastids. Only the first two steps of the plastidial pathway, which involve the condensation of pyruvate and glyceraldehyde 3-phosphate to deoxyxylulose 5-phosphate followed by intramolecular rearrangement and reduction to 2-Cmethylerythritol 4-phosphate, have been established. Here we report the cloning from peppermint (Mentha ؋ piperita) and E. coli, and expression, of a kinase that catalyzes the phosphorylation of isopentenyl monophosphate as the last step of this biosynthetic sequence to isopentenyl diphosphate. The plant gene defines an ORF of 1,218 bp that, when the proposed plastidial targeting sequence is excluded, corresponds to Ϸ308 aa with a mature size of Ϸ33 kDa. The E. coli gene (ychB), which is located at 27.2 min of the chromosomal map, consists of 852 nt, encoding a deduced enzyme of 283 aa with a size of 31 kDa. These enzymes represent a conserved class of the GHMP family of kinases, which includes galactokinase, homoserine kinase, mevalonate kinase, and phosphomevalonate kinase, with homologues in plants and several eubacteria. Besides the preferred substrate isopentenyl monophosphate, the recombinant peppermint and E. coli kinases also phosphorylate isopentenol, and, much less efficiently, dimethylallyl alcohol, but dimethylallyl monophosphate does not serve as a substrate. Incubation of secretory cells isolated from peppermint glandular trichomes with isopentenyl monophosphate resulted in the rapid production of monoterpenes and sesquiterpenes, confirming that isopentenyl monophosphate is the physiologically relevant, terminal intermediate of the deoxyxylulose 5-phosphate pathway.deoxyxylulose 5-phosphate ͉ isoprenoid biosynthesis

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Section: Cloning and Heterologous Expression Of A Putative Kinase Frommentioning

confidence: 99%

“…The enzyme that catalyzes this reaction belongs to a family of transketolases, and the corresponding gene has been isolated from Escherichia coli (15,16), peppermint (17), and pepper (18). In the second step of this pathway, rearrangement and reduction of DXP yield 2-C-methylerythritol 4-phosphate (MEP) (19)(20)(21) (Fig. 1).…”

mentioning

confidence: 99%

Isopentenyl diphosphate biosynthesis via a mevalonate-independent pathway: Isopentenyl monophosphate kinase catalyzes the terminal enzymatic step

Croteau

1999

Proc. Natl. Acad. Sci. U.S.A.

103

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“…strain CL190 (7). In the second step, the intramolecular rearrangement of DXP was assumed to give a hypothetical rearrangement product, 2-C-methylerythrose 4-phosphate, which was then converted to 2-C-methyl-D-erythritol 4-phosphate (MEP) by an unspecified reduction process (8,9). We have succeeded in the first cloning and overexpression of the gene (dxr, formerly yaeM) encoding the DXP reductoisomerase from E. coli and have shown that the recombinant enzyme catalyzed the formation of MEP from DXP in a single step in the presence of both NADPH and a divalent cation, such as Mn 2ϩ , Mg 2ϩ , or Co 2ϩ (10,11).…”

mentioning

confidence: 99%

Characterization of 1-Deoxy-d-xylulose 5-Phosphate Reductoisomerase, an Enzyme Involved in Isopentenyl Diphosphate Biosynthesis, and Identification of Its Catalytic Amino Acid Residues

Kuzuyama¹,

Takahashi²,

Takagi

et al. 2000

Journal of Biological Chemistry

106

104

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1-Deoxy-D-xylulose 5-phosphate (DXP) reductoisomerase, which simultaneously catalyzes the intramolecular rearrangement and reduction of DXP to form 2-C-methyl-D-erythritol 4-phosphate, constitutes a key enzyme of an alternative mevalonate-independent pathway for isopentenyl diphosphate biosynthesis. The dxr gene encoding this enzyme from Escherichia coli was overexpressed as a histidine-tagged protein and characterized in detail. DNA sequencing analysis of the dxr genes from 10 E. coli dxr-deficient mutants revealed base substitution mutations at four points: two nonsense mutations and two amino acid substitutions (Gly 14 to Asp 14 and Glu 231 to Lys 231 ). Diethyl pyrocarbonate treatment inactivated DXP reductoisomerase, and subsequent hydroxylamine treatment restored the activity of the diethyl pyrocarbonate-treated enzyme. To characterize these defects, we overexpressed the mutant enzymes G14D, E231K, H153Q, H209Q, and H257Q. All of these mutant enzymes except for G14D were obtained as soluble proteins. Although the purified enzyme E231K had wildtype K m values for DXP and NADPH, the mutant enzyme had less than a 0.24% wild-type k cat value. K m values of H153Q, H209Q, and H257Q for DXP increased to 3.5-, 7.6-, and 19-fold the wild-type value, respectively. These results indicate that Glu 231 of E. coli DXP reductoisomerase plays an important role(s) in the conversion of DXP to 2-C-methyl-D-erythritol 4-phosphate, and that His 153

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“…A facile route to ME was reported by Duvold et al in 1997, wherein ME was produced in 85% yield (80% e.e.) from 3-methylfuran-2(5H)-one 36 or 25% from citraconic anhydride 37 in four steps [54]. Sharpless asymmetric dihydroxylation was used to introduce the C-2 and C-3 hydroxyl groups with the correct stereochemistry into cis-olefin 38 to provide the diacetate 39 protected ME in excellent yield.…”

Section: Syntheses Of 2c-methyl-d-erythritol 40 (Me)-mentioning

confidence: 99%

Bioconversion of Lignocellulose: Inhibitors and Detoxifi cation

2015

New Biotechnologies for Increased Energy Security

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Biosynthesis of , a putative C5 intermediate in the mevalonate independent pathway for isoprenoid biosynthesis

Cited by 92 publications

References 18 publications

Isopentenyl diphosphate biosynthesis via a mevalonate-independent pathway: Isopentenyl monophosphate kinase catalyzes the terminal enzymatic step

Isopentenyl diphosphate biosynthesis via a mevalonate-independent pathway: Isopentenyl monophosphate kinase catalyzes the terminal enzymatic step

Characterization of 1-Deoxy-d-xylulose 5-Phosphate Reductoisomerase, an Enzyme Involved in Isopentenyl Diphosphate Biosynthesis, and Identification of Its Catalytic Amino Acid Residues

Bioconversion of Lignocellulose: Inhibitors and Detoxifi cation

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