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Literature summary for 1.1.1.267 extracted from

  • Fox, D.T.; Poulter, C.D.
    Mechanistic studies with 2-C-methyl-D-erythritol 4-phosphate synthase from Escherichia coli (2005), Biochemistry, 44, 8360-8368.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
overexpression in strain BL21 as His-tagged enzyme Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
additional information 1,1,1-trifluoro-1-deoxy-D-xylulose 5-phosphoric acid, 1,1-difluoro-1-deoxy-D-xylulose 5-phosphoric acid, and 1,2-dideoxy-D-hexulose 6-phosphate are poor inhibitors, most likely because of the increase in steric bulk at C1 Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information steady-state and single-turnover kinetics, primary kinetic isotope effects, overview Escherichia coli
0.0005
-
NADPH pH 7.6, 37°C, recombinant enzyme Escherichia coli
0.045
-
1-deoxy-D-xylulose 5-phosphate pH 7.6, 37°C, recombinant enzyme Escherichia coli
0.227
-
1-fluoro-1-deoxy-D-xylulose 5-phosphate pH 7.6, 37°C, recombinant enzyme Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ activates Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1-deoxy-D-xylulose 5-phosphate + NADPH + H+ Escherichia coli
-
2-C-methyl-D-erythritol 4-phosphate + NADP+
-
r

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from overexpressing strain BL21 by nickel affinity chromatography Escherichia coli

Reaction

Reaction Comment Organism Reaction ID
2-C-methyl-D-erythritol 4-phosphate + NADP+ = 1-deoxy-D-xylulose 5-phosphate + NADPH + H+ the reaction does not exhibit burst kinetics for either substrate, indicating that product release is not rate-limiting, the rearrangement step, which precedes hydride transfer, is rate-limiting for 1-deoxy-D-xylulose 5-phosphate but becomes partially ratelimiting for 1-fluoro-1-deoxy-D-xylulose 5-phosphate, thus, rearrangement appears to be enhanced by substitution of a hydrogen atom in the methyl group of 1-deoxy-D-xylulose 5-phosphate by fluorine, consistent with a retro-aldol/aldol mechanism for the rearrangement during conversion of 1-deoxy-D-xylulose 5-phosphate to 2-C-methyl-D-erythritol 4-phosphate Escherichia coli

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
-
Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1-deoxy-D-xylulose 5-phosphate + NADPH + H+
-
Escherichia coli 2-C-methyl-D-erythritol 4-phosphate + NADP+
-
r
1-deoxy-D-xylulose 5-phosphate + NADPH + H+ i.e. DXP Escherichia coli 2-C-methyl-D-erythritol 4-phosphate + NADP+
-
r
1-fluoro-1-deoxy-D-xylulose 5-phosphate + NADPH i.e. CF-DXP, results in about 25% of the activity with 1-deoxy-D-xylulose 5-phosphate Escherichia coli ?
-
r
additional information 1,1,1-trifluoro-1-deoxy-D-xylulose 5-phosphoric acid, 1,1-difluoro-1-deoxy-D-xylulose 5-phosphoric acid, and 1,2-dideoxy-D-hexulose 6-phosphate are poor substrates Escherichia coli ?
-
?

Synonyms

Synonyms Comment Organism
2-C-methyl-D-erythritol 4-phosphate synthase
-
Escherichia coli
MEP synthase
-
Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
29
-
1-deoxy-D-xylulose 5-phosphate pH 7.6, 37°C, recombinant enzyme Escherichia coli
37
-
1-fluoro-1-deoxy-D-xylulose 5-phosphate pH 7.6, 37°C, recombinant enzyme Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.6
-
assay at Escherichia coli

Cofactor

Cofactor Comment Organism Structure
NADP+
-
Escherichia coli
NADPH
-
Escherichia coli