Identification and characterization of (1R,6R)-2-succinyl-6-hydroxy-2,4-cyclohexadiene-1-carboxylate synthase in the menaquinone biosynthesis of Escherichia coli
Jiang, M.; Chen, X.; Guo, Z.F.; Cao, Y.; Chen, M.; Guo, Z.; Biochemistry 47, 3426-3434 (2008) 
Data extracted from this reference:
Engineering
EC Number
Protein Variants
Commentary
Organism
4.2.99.20
D210A
decreased activity; kcat/Km is 9091fold lower than wild-type value
Escherichia coli
4.2.99.20
H232A
decreased activity; kcat/Km is 208333fold lower than wild-type value
Escherichia coli
4.2.99.20
S86A
decreased activity; kcat/Km is 1498fold lower than wild-type value
Escherichia coli
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.2.99.20
0.0083
-
2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
wild-type at 37°C
Escherichia coli
4.2.99.20
0.0083
-
5-enolpyruvoyl-6-hydroxy-2-succinyl-cyclohex-3-ene-1-carboxylate
pH 7.0, 37°C
Escherichia coli
4.2.99.20
0.0086
-
2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant H232A
Escherichia coli
4.2.99.20
0.0086
-
5-enolpyruvoyl-6-hydroxy-2-succinyl-cyclohex-3-ene-1-carboxylate
25°C, mutant enzyme H232A
Escherichia coli
4.2.99.20
0.0101
-
2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
wild-type at 25°C
Escherichia coli
4.2.99.20
0.0191
-
5-enolpyruvoyl-6-hydroxy-2-succinyl-cyclohex-3-ene-1-carboxylate
25°C, wild-type enzyme
Escherichia coli
4.2.99.20
0.028
-
2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant S86A at 25°C
Escherichia coli
4.2.99.20
0.028
-
5-enolpyruvoyl-6-hydroxy-2-succinyl-cyclohex-3-ene-1-carboxylate
25°C, mutant enzyme S86A
Escherichia coli
4.2.99.20
0.118
-
2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant D210A at 25°C
Escherichia coli
4.2.99.20
0.118
-
5-enolpyruvoyl-6-hydroxy-2-succinyl-cyclohex-3-ene-1-carboxylate
25°C, mutant enzyme D210A
Escherichia coli
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
4.2.99.20
additional information
SHCHC synthase activity of MenH is not affected by the addition of a divalent ion (1 mM of Mg2+, Ba2+, Mn2+, Ca2+, Co2+, or Ni2+)
Escherichia coli
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
4.2.99.20
25400
-
gel filtration
Escherichia coli
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
4.2.99.20
5-enolpyruvoyl-6-hydroxy-2-succinyl-cyclohex-3-ene-1-carboxylate
Escherichia coli
step in menaquinone biosynthesis
(1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate + pyruvate
-
-
?
Organism
EC Number
Organism
UniProt
Commentary
Textmining
4.2.99.20
Escherichia coli
-
-
-
Purification (Commentary)
EC Number
Purification (Commentary)
Organism
4.2.99.20
; combination of Ni2+-affinity chromatography and size-exclusion chromatography
Escherichia coli
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
4.2.99.20
2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
assay at 37°C, pH 7.0
685225
Escherichia coli
(1R,6R)-2-succinyl-6-hydroxy-2,4-cyclohexadiene-1-carboxylate + pyruvate
-
-
-
?
4.2.99.20
5-enolpyruvoyl-6-hydroxy-2-succinyl-cyclohex-3-ene-1-carboxylate
-
685225
Escherichia coli
(1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate + pyruvate
-
-
-
?
4.2.99.20
5-enolpyruvoyl-6-hydroxy-2-succinyl-cyclohex-3-ene-1-carboxylate
step in menaquinone biosynthesis
685225
Escherichia coli
(1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate + pyruvate
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
4.2.99.20
monomer
; 1 * 25400
Escherichia coli
Synonyms
EC Number
Synonyms
Commentary
Organism
4.2.99.20
(1R,6R)-2-succinyl-6-hydroxy-2,4-cyclohexadiene-1-carboxylate synthase
-
Escherichia coli
4.2.99.20
MenH
-
Escherichia coli
4.2.99.20
SHCHC synthase
-
Escherichia coli
4.2.99.20
YfbB
-
Escherichia coli
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
4.2.99.20
37
-
assay at
Escherichia coli
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
4.2.99.20
0.18
-
5-enolpyruvoyl-6-hydroxy-2-succinyl-cyclohex-3-ene-1-carboxylate
25°C, mutant enzyme D210A
Escherichia coli
4.2.99.20
0.29
-
5-enolpyruvoyl-6-hydroxy-2-succinyl-cyclohex-3-ene-1-carboxylate
25°C, mutant enzyme S86A
Escherichia coli
4.2.99.20
5.8
-
5-enolpyruvoyl-6-hydroxy-2-succinyl-cyclohex-3-ene-1-carboxylate
25°C, mutant enzyme H232A
Escherichia coli
4.2.99.20
147
-
5-enolpyruvoyl-6-hydroxy-2-succinyl-cyclohex-3-ene-1-carboxylate
25°C, wild-type enzyme
Escherichia coli
4.2.99.20
167
-
5-enolpyruvoyl-6-hydroxy-2-succinyl-cyclohex-3-ene-1-carboxylate
pH 7.0, 37°C
Escherichia coli
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
4.2.99.20
7
-
assay at
Escherichia coli
Engineering (protein specific)
EC Number
Protein Variants
Commentary
Organism
4.2.99.20
D210A
decreased activity; kcat/Km is 9091fold lower than wild-type value
Escherichia coli
4.2.99.20
H232A
decreased activity; kcat/Km is 208333fold lower than wild-type value
Escherichia coli
4.2.99.20
S86A
decreased activity; kcat/Km is 1498fold lower than wild-type value
Escherichia coli
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.2.99.20
0.0083
-
2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
wild-type at 37°C
Escherichia coli
4.2.99.20
0.0083
-
5-enolpyruvoyl-6-hydroxy-2-succinyl-cyclohex-3-ene-1-carboxylate
pH 7.0, 37°C
Escherichia coli
4.2.99.20
0.0086
-
2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant H232A
Escherichia coli
4.2.99.20
0.0086
-
5-enolpyruvoyl-6-hydroxy-2-succinyl-cyclohex-3-ene-1-carboxylate
25°C, mutant enzyme H232A
Escherichia coli
4.2.99.20
0.0101
-
2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
wild-type at 25°C
Escherichia coli
4.2.99.20
0.0191
-
5-enolpyruvoyl-6-hydroxy-2-succinyl-cyclohex-3-ene-1-carboxylate
25°C, wild-type enzyme
Escherichia coli
4.2.99.20
0.028
-
2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant S86A at 25°C
Escherichia coli
4.2.99.20
0.028
-
5-enolpyruvoyl-6-hydroxy-2-succinyl-cyclohex-3-ene-1-carboxylate
25°C, mutant enzyme S86A
Escherichia coli
4.2.99.20
0.118
-
2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant D210A at 25°C
Escherichia coli
4.2.99.20
0.118
-
5-enolpyruvoyl-6-hydroxy-2-succinyl-cyclohex-3-ene-1-carboxylate
25°C, mutant enzyme D210A
Escherichia coli
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
4.2.99.20
additional information
SHCHC synthase activity of MenH is not affected by the addition of a divalent ion (1 mM of Mg2+, Ba2+, Mn2+, Ca2+, Co2+, or Ni2+)
Escherichia coli
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
4.2.99.20
25400
-
gel filtration
Escherichia coli
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
4.2.99.20
5-enolpyruvoyl-6-hydroxy-2-succinyl-cyclohex-3-ene-1-carboxylate
Escherichia coli
step in menaquinone biosynthesis
(1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate + pyruvate
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
4.2.99.20
; combination of Ni2+-affinity chromatography and size-exclusion chromatography
Escherichia coli
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
4.2.99.20
2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
assay at 37°C, pH 7.0
685225
Escherichia coli
(1R,6R)-2-succinyl-6-hydroxy-2,4-cyclohexadiene-1-carboxylate + pyruvate
-
-
-
?
4.2.99.20
5-enolpyruvoyl-6-hydroxy-2-succinyl-cyclohex-3-ene-1-carboxylate
-
685225
Escherichia coli
(1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate + pyruvate
-
-
-
?
4.2.99.20
5-enolpyruvoyl-6-hydroxy-2-succinyl-cyclohex-3-ene-1-carboxylate
step in menaquinone biosynthesis
685225
Escherichia coli
(1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate + pyruvate
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
4.2.99.20
monomer
; 1 * 25400
Escherichia coli
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
4.2.99.20
37
-
assay at
Escherichia coli
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
4.2.99.20
0.18
-
5-enolpyruvoyl-6-hydroxy-2-succinyl-cyclohex-3-ene-1-carboxylate
25°C, mutant enzyme D210A
Escherichia coli
4.2.99.20
0.29
-
5-enolpyruvoyl-6-hydroxy-2-succinyl-cyclohex-3-ene-1-carboxylate
25°C, mutant enzyme S86A
Escherichia coli
4.2.99.20
5.8
-
5-enolpyruvoyl-6-hydroxy-2-succinyl-cyclohex-3-ene-1-carboxylate
25°C, mutant enzyme H232A
Escherichia coli
4.2.99.20
147
-
5-enolpyruvoyl-6-hydroxy-2-succinyl-cyclohex-3-ene-1-carboxylate
25°C, wild-type enzyme
Escherichia coli
4.2.99.20
167
-
5-enolpyruvoyl-6-hydroxy-2-succinyl-cyclohex-3-ene-1-carboxylate
pH 7.0, 37°C
Escherichia coli
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
4.2.99.20
7
-
assay at
Escherichia coli
General Information
EC Number
General Information
Commentary
Organism
4.2.99.20
physiological function
involved in menaquinone biosynthesis
Escherichia coli
General Information (protein specific)
EC Number
General Information
Commentary
Organism
4.2.99.20
physiological function
involved in menaquinone biosynthesis
Escherichia coli
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
4.2.99.20
67
-
2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant H232A
Escherichia coli
4.2.99.20
1500
-
2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant D210A at 25°C
Escherichia coli
4.2.99.20
10000
-
2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant S86A at 25°C
Escherichia coli
4.2.99.20
14000000
-
2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
wild-type at 25°C
Escherichia coli
4.2.99.20
20000000
-
2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
wild-type at 37°C
Escherichia coli
KCat/KM [mM/s] (protein specific)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
4.2.99.20
67
-
2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant H232A
Escherichia coli
4.2.99.20
1500
-
2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant D210A at 25°C
Escherichia coli
4.2.99.20
10000
-
2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
mutant S86A at 25°C
Escherichia coli
4.2.99.20
14000000
-
2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
wild-type at 25°C
Escherichia coli
4.2.99.20
20000000
-
2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
wild-type at 37°C
Escherichia coli