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.

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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

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Release 2023.1 (January 2023)