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Literature summary extracted from

  • Zepeck, F.; Graewert, T.; Kaiser, J.; Schramek, N.; Eisenreich, W.; Bacher, A.; Rohdich, F.
    Biosynthesis of isoprenoids. Purification and properties of IspG protein from Escherichia coli (2005), J. Org. Chem., 70, 9168-9174.
    View publication on PubMed

Protein Variants

EC Number Protein Variants Comment Organism
1.17.7.3 C270S the replacement of any of the three conserved cysteine residues affords mutant proteins which are devoid of catalytic activity and contain less than 6% of Fe2+ and less than 23% of S2- as compared to the wild-type protein Escherichia coli
1.17.7.3 C273S the replacement of any of the three conserved cysteine residues affords mutant proteins which are devoid of catalytic activity and contain less than 6% of Fe2+ and less than 23% of S2- as compared to the wild-type protein Escherichia coli
1.17.7.3 C306S the replacement of any of the three conserved cysteine residues affords mutant proteins which are devoid of catalytic activity and contain less than 6% of Fe2+ and less than 23% of S2- as compared to the wild-type protein Escherichia coli

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.17.7.3 Ca2+ 1 mM, 43% inhibition Escherichia coli
1.17.7.3 Co2+ 1 mM, 29% inhibition Escherichia coli
1.17.7.3 EDTA 10 mM, complete inhibition, reactivated by Mn2+ to a level of 25% Escherichia coli
1.17.7.3 Fe2+ 1 mM, 11% inhibition Escherichia coli
1.17.7.3 Mg2+ 1 mM, 25% inhibition Escherichia coli
1.17.7.3 Ni2+ 1 mM, 30% inhibition Escherichia coli
1.17.7.3 Zn2+ 1 mM, 98% inhibition Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.17.7.3 0.7
-
 2-C-methyl-D-erythritol 2,4-cyclodiphosphate pH 8.0, 37°C, photometric assay Escherichia coli

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.17.7.3 iron-sulfur centre the protein contained 2.4 iron ions and 4.4 sulfide ions per subunit Escherichia coli
1.17.7.3 Mn2+ reactivates EDTA-inactivated enzyme Escherichia coli
1.17.7.3 additional information the enzyme is active in absence of added metal ions Escherichia coli

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.17.7.3 84000
-
 x * 84000, SDS-PAGE Escherichia coli

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.17.7.3 2-C-methyl-D-erythritol 2,4-cyclodiphosphate + reduced flavodoxin Escherichia coli the enzyme is involved in the nonmevalonate pathway of isoprenoid biosynthesis (E)-4-hydroxy-3-methylbut-2-en-1-yl diphosphate + H2O + oxidized flavodoxin
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.17.7.3 Escherichia coli P62620
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.17.7.3 the recombinant MalE/IspG fusion protein (expression level of about 40% with respect to the total soluble cell protein) is purified by affinity chromatography on an amylose resin FF column that is developed with a gradient of 0-10 mM maltose under anaerobic conditions, whereby the enzyme is enriched by a factor of 2.4 Escherichia coli

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.17.7.3 0.045
-
 activity with 2-C-methyl-D-erythritol 2,4-cyclodiphosphate and reduced deazaflavin, pH 8.0, 37°C Escherichia coli
1.17.7.3 0.099
-
 activity with 2-C-methyl-D-erythritol 2,4-cyclodiphosphate and reduced flavodoxin, pH 8.0, 37°C, photometric assay Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.17.7.3 2-C-methyl-D-erythritol 2,4-cyclodiphosphate + reduced deazaflavin low activity with photoreduced deazaflavin as an artifactual electron donor Escherichia coli (E)-4-hydroxy-3-methylbut-2-en-1-yl diphosphate + H2O + oxidized deazaflavin
-
 ?
1.17.7.3 2-C-methyl-D-erythritol 2,4-cyclodiphosphate + reduced flavodoxin the enzyme is involved in the nonmevalonate pathway of isoprenoid biosynthesis Escherichia coli (E)-4-hydroxy-3-methylbut-2-en-1-yl diphosphate + H2O + oxidized flavodoxin
-
 ?
1.17.7.3 2-C-methyl-D-erythritol 2,4-cyclodiphosphate + reduced flavodoxin lower apparent activities are found using photoreduced deazaflavin as an artifactual electron donor, whereas dithionite is unable to serve as an artificial electron donor Escherichia coli (E)-4-hydroxy-3-methylbut-2-en-1-yl diphosphate + H2O + oxidized flavodoxin
-
 ?

Subunits

EC Number Subunits Comment Organism
1.17.7.3 ? x * 84000, SDS-PAGE Escherichia coli

Synonyms

EC Number Synonyms Comment Organism
1.17.7.3 IspG ambiguous Escherichia coli

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.17.7.3 37
-
 assay at Escherichia coli

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.17.7.3 9
-
-
 Escherichia coli

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.17.7.3 8 9.5 pH 8.0: about 65% of maximal activity, pH 9.5: about 75% of maximal activity Escherichia coli

Cofactor

EC Number Cofactor Comment Organism Structure
1.17.7.3 [4Fe-4S]-center the protein contains 2.4 iron ions and 4.4 sulfide ions per subunit Escherichia coli

General Information

EC Number General Information Comment Organism
1.17.7.3 metabolism the enzyme is involved in the nonmevalonate pathway of isoprenoid biosynthesis Escherichia coli