BRENDA - Enzyme Database
show all sequences of 4.2.99.20

Catalytic mechanism of SHCHC synthase in the menaquinone biosynthesis of Escherichia coli: identification and mutational analysis of the active site residues

Jiang, M.; Chen, X.; Wu, X.H.; Chen, M.; Wu, Y.D.; Guo, Z.; Biochemistry 48, 6921-6931 (2009)

Data extracted from this reference:

Cloned(Commentary)
Cloned (Commentary)
Organism
expressed in Escherichia coli BL21(DE3)
Escherichia coli
Engineering
Protein Variants
Commentary
Organism
D128A
conserved among MenH proteins
Escherichia coli
D210A
part of the catalytically essential triad
Escherichia coli
H232A
part of the catalytically essential triad
Escherichia coli
H232K
part of the catalytically essential triad
Escherichia coli
K212A
conserved among MenH proteins
Escherichia coli
R124A
conserved among MenH proteins
Escherichia coli
R168A
conserved among MenH proteins
Escherichia coli
R90A
conserved among MenH proteins
Escherichia coli
S86A
part of the catalytically essential triad
Escherichia coli
W147A
conserved among MenH proteins
Escherichia coli
W147F
conserved among MenH proteins
Escherichia coli
Y85F
conserved among MenH proteins
Escherichia coli
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0086
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
H232A mutant protein
Escherichia coli
0.01
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
wild-type protein
Escherichia coli
0.014
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
Y85F mutant protein
Escherichia coli
0.0194
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
K212A mutant protein
Escherichia coli
0.0237
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
W147A mutant protein
Escherichia coli
0.028
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
S86A mutant protein
Escherichia coli
0.074
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
W147F mutant protein
Escherichia coli
0.084
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
R124A mutant protein
Escherichia coli
0.085
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
D128A mutant protein
Escherichia coli
0.118
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
D210A mutant protein
Escherichia coli
0.134
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
R168A mutant protein
Escherichia coli
0.159
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
R90A mutant protein
Escherichia coli
0.532
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
H232K mutant protein
Escherichia coli
Organism
Organism
UniProt
Commentary
Textmining
Escherichia coli
-
-
-
Purification (Commentary)
Purification (Commentary)
Organism
immobilized metal ion affinity chromatography, gel filtration
Escherichia coli
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
-
702326
Escherichia coli
(1R,6R)-2-succinyl-6-hydroxy-2,4-cyclohexadiene-1-carboxylate + pyruvate
-
-
-
?
Synonyms
Synonyms
Commentary
Organism
(1R,6R)-2-succinyl-6-hydroxy-2,4-cyclohexadiene-1-carboxylate synthase
-
Escherichia coli
MenH
-
Escherichia coli
SHCHC
-
Escherichia coli
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.00058
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
H232A mutant protein
Escherichia coli
0.021
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
H232K mutant protein
Escherichia coli
0.047
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
R90A mutant protein
Escherichia coli
0.056
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
W147A mutant protein
Escherichia coli
0.18
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
D210A mutant protein
Escherichia coli
0.29
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
S86A mutant protein
Escherichia coli
0.51
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
R124A mutant protein
Escherichia coli
2.58
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
Y85F mutant protein
Escherichia coli
2.9
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
R168A mutant protein
Escherichia coli
15.4
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
W147F mutant protein
Escherichia coli
41.5
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
K212A mutant protein
Escherichia coli
115
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
D128A mutant protein
Escherichia coli
147
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
wild-type protein
Escherichia coli
Cloned(Commentary) (protein specific)
Commentary
Organism
expressed in Escherichia coli BL21(DE3)
Escherichia coli
Engineering (protein specific)
Protein Variants
Commentary
Organism
D128A
conserved among MenH proteins
Escherichia coli
D210A
part of the catalytically essential triad
Escherichia coli
H232A
part of the catalytically essential triad
Escherichia coli
H232K
part of the catalytically essential triad
Escherichia coli
K212A
conserved among MenH proteins
Escherichia coli
R124A
conserved among MenH proteins
Escherichia coli
R168A
conserved among MenH proteins
Escherichia coli
R90A
conserved among MenH proteins
Escherichia coli
S86A
part of the catalytically essential triad
Escherichia coli
W147A
conserved among MenH proteins
Escherichia coli
W147F
conserved among MenH proteins
Escherichia coli
Y85F
conserved among MenH proteins
Escherichia coli
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0086
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
H232A mutant protein
Escherichia coli
0.01
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
wild-type protein
Escherichia coli
0.014
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
Y85F mutant protein
Escherichia coli
0.0194
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
K212A mutant protein
Escherichia coli
0.0237
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
W147A mutant protein
Escherichia coli
0.028
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
S86A mutant protein
Escherichia coli
0.074
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
W147F mutant protein
Escherichia coli
0.084
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
R124A mutant protein
Escherichia coli
0.085
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
D128A mutant protein
Escherichia coli
0.118
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
D210A mutant protein
Escherichia coli
0.134
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
R168A mutant protein
Escherichia coli
0.159
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
R90A mutant protein
Escherichia coli
0.532
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
H232K mutant protein
Escherichia coli
Purification (Commentary) (protein specific)
Commentary
Organism
immobilized metal ion affinity chromatography, gel filtration
Escherichia coli
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
-
702326
Escherichia coli
(1R,6R)-2-succinyl-6-hydroxy-2,4-cyclohexadiene-1-carboxylate + pyruvate
-
-
-
?
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.00058
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
H232A mutant protein
Escherichia coli
0.021
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
H232K mutant protein
Escherichia coli
0.047
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
R90A mutant protein
Escherichia coli
0.056
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
W147A mutant protein
Escherichia coli
0.18
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
D210A mutant protein
Escherichia coli
0.29
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
S86A mutant protein
Escherichia coli
0.51
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
R124A mutant protein
Escherichia coli
2.58
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
Y85F mutant protein
Escherichia coli
2.9
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
R168A mutant protein
Escherichia coli
15.4
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
W147F mutant protein
Escherichia coli
41.5
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
K212A mutant protein
Escherichia coli
115
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
D128A mutant protein
Escherichia coli
147
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
wild-type protein
Escherichia coli
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.0392
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
H232K mutant protein
Escherichia coli
0.067
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
H232A mutant protein
Escherichia coli
0.295
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
R90A mutant protein
Escherichia coli
1.5
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
D210A mutant protein
Escherichia coli
2.4
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
W147A mutant protein
Escherichia coli
6.1
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
R124A mutant protein
Escherichia coli
10
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
S86A mutant protein
Escherichia coli
15
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
R168A mutant protein
Escherichia coli
180
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
Y85F mutant protein
Escherichia coli
220
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
W147F mutant protein
Escherichia coli
1400
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
D128A mutant protein
Escherichia coli
2200
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
K212A mutant protein
Escherichia coli
14000
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
wild-type protein
Escherichia coli
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.0392
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
H232K mutant protein
Escherichia coli
0.067
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
H232A mutant protein
Escherichia coli
0.295
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
R90A mutant protein
Escherichia coli
1.5
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
D210A mutant protein
Escherichia coli
2.4
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
W147A mutant protein
Escherichia coli
6.1
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
R124A mutant protein
Escherichia coli
10
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
S86A mutant protein
Escherichia coli
15
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
R168A mutant protein
Escherichia coli
180
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
Y85F mutant protein
Escherichia coli
220
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
W147F mutant protein
Escherichia coli
1400
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
D128A mutant protein
Escherichia coli
2200
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
K212A mutant protein
Escherichia coli
14000
-
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate
wild-type protein
Escherichia coli
Other publictions for EC
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Synonyms
Temperature Optimum [C]
Temperature Range [C]
Temperature Stability [C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [C] (protein specific)
Temperature Range [C] (protein specific)
Temperature Stability [C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
730042
Sun
Molecular basis of the general ...
Escherichia coli
J. Biol. Chem.
289
15867-15879
2014
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1
1
7
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6
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1
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1
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1
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1
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5
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1
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1
7
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6
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1
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5
-
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-
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6
6
730729
Johnston
Crystal structures of E. coli ...
Escherichia coli
PLoS ONE
8
e61325
2013
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1
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2
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1
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714632
Dawson
Exploiting the high-resolution ...
Staphylococcus aureus
BMC Struct. Biol.
11
19
2011
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1
1
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1
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1
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1
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1
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-
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702326
Jiang
Catalytic mechanism of SHCHC s ...
Escherichia coli
Biochemistry
48
6921-6931
2009
-
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1
-
12
-
-
13
-
-
-
-
-
1
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1
-
-
-
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1
-
3
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13
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-
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-
-
1
-
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12
-
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13
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1
-
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-
1
-
-
-
-
13
-
-
-
-
-
-
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-
13
13
685225
Jiang
Identification and characteriz ...
Escherichia coli
Biochemistry
47
3426-3434
2008
-
-
-
-
3
-
-
10
-
1
1
1
-
4
-
-
1
-
-
-
-
-
3
1
4
1
-
-
5
1
-
-
-
-
-
-
-
-
-
-
-
3
-
-
-
-
10
-
1
1
1
-
-
-
1
-
-
-
-
3
1
1
-
-
5
1
-
-
-
-
1
1
-
5
5
685086
Jiang
Menaquinone biosynthesis in Es ...
Escherichia coli K-12
Biochemistry
46
10979-10989
2007
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-
-
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-
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1
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3
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2
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1
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1
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2
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