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Literature summary for 1.17.7.3 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
Search for more literature for 1.17.7.3

Protein Variants

Protein Variants Comment Organism
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
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
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

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

KM Value [mM]

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

Metals/Ions

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

Molecular Weight [Da]

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

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
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

Organism UniProt Comment Textmining
Escherichia coli P62620
-
-

Purification (Commentary)

Purification (Comment) Organism
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]

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

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
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
-
 ?
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
-
 ?
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

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

Synonyms

Synonyms Comment Organism
IspG ambiguous Escherichia coli

Temperature Optimum [°C]

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

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9
-
-
 Escherichia coli

pH Range

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

Cofactor

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

General Information

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