BRENDA - Enzyme Database show
show all sequences of 4.1.3.17

Structural and kinetic characterization of 4-hydroxy-4-methyl-2-oxoglutarate/4-carboxy-4-hydroxy-2-oxoadipate aldolase, a protocatechuate degradation enzyme evolutionarily convergent with the HpaI and DmpG pyruvate aldolases

Wang, W.; Mazurkewich, S.; Kimber, M.S.; Seah, S.Y.; J. Biol. Chem. 285, 36608-36615 (2010)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
expressed in Escherichia coli BL21(lambdaDE3)
Pseudomonas putida
Crystallization (Commentary)
Crystallization
Organism
hanging drop method, PEG 4000
Pseudomonas putida
Engineering
Amino acid exchange
Commentary
Organism
R123A
no activity
Pseudomonas putida
R123K
retains some activity
Pseudomonas putida
Inhibitors
Inhibitors
Commentary
Organism
Structure
oxalate
competitive inhibitor
Pseudomonas putida
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.022
-
4-hydroxy-4-methyl-2-oxoglutarate
wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C
Pseudomonas putida
0.036
-
oxaloacetate
wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C; substrate inhibition constante 0.25 mM
Pseudomonas putida
0.071
-
2-hydroxy-4-oxopentanedioic acid
wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C
Pseudomonas putida
0.15
-
2-hydroxy-4-oxopentanedioic acid
wild type protein, 1.0 mM Mg2+, pH 8.0, 25°C
Pseudomonas putida
0.26
-
4-hydroxy-4-methyl-2-oxoglutarate
wild type protein, 1.0 mM Mg2+, pH 8.0, 25°C
Pseudomonas putida
0.3
-
oxaloacetate
wild type protein, 1.0 mM Mg2+, pH 8.0, 25°C; substrate inhibition constante 0.37 mM
Pseudomonas putida
5.5
-
4-hydroxy-4-methyl-2-oxoglutarate
R123K mutant protein, pH 8.0, 25°C
Pseudomonas putida
8.8
-
4-hydroxy-2-oxopentanoate
wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C
Pseudomonas putida
25
-
4-hydroxy-2-oxopentanoate
wild type protein, 1.0 mM Mg2+, pH 8.0, 25°C
Pseudomonas putida
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Mg2+
Mn2+: 90.3% activity, Co2+: 64.7% activity, Zn2+: 49.6% activity
Pseudomonas putida
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
26000
-
6 * 26000, crystal structure, gel filtration and SDS-PAGE
Pseudomonas putida
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Pseudomonas putida
-
-
-
Purification (Commentary)
Commentary
Organism
ion exchange chromatography, hydrophobic interaction chromatography, gel filtration
Pseudomonas putida
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
37.7
-
wild type protein, pH 8.0, 25°C
Pseudomonas putida
Storage Stability
Storage Stability
Organism
-80°C, 20 mM sodium HEPES buffer, pH7.5, stable for 2 months
Pseudomonas putida
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2-hydroxy-4-oxopentanedioic acid
-
715556
Pseudomonas putida
?
-
-
-
?
4-hydroxy-2-oxopentanoate
-
715556
Pseudomonas putida
?
-
-
-
?
4-hydroxy-4-methyl-2-oxoglutarate
-
715556
Pseudomonas putida
2 pyruvate
-
-
-
r
oxaloacetate
-
715556
Pseudomonas putida
?
-
-
-
?
Subunits
Subunits
Commentary
Organism
homohexamer
6 * 26000, crystal structure, gel filtration and SDS-PAGE
Pseudomonas putida
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.048
-
4-hydroxy-2-oxopentanoate
wild type protein, 1.0 mM Mg2+, pH 8.0, 25°C
Pseudomonas putida
0.058
-
2-hydroxy-4-oxopentanedioic acid
wild type protein, 1.0 mM Mg2+, pH 8.0, 25°C
Pseudomonas putida
0.099
-
4-hydroxy-4-methyl-2-oxoglutarate
R123K mutant protein, pH 8.0, 25°C
Pseudomonas putida
0.19
-
4-hydroxy-2-oxopentanoate
wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C
Pseudomonas putida
0.25
-
2-hydroxy-4-oxopentanedioic acid
wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C
Pseudomonas putida
1.52
-
oxaloacetate
wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C
Pseudomonas putida
1.85
-
oxaloacetate
wild type protein, 1.0 mM Mg2+, pH 8.0, 25°C
Pseudomonas putida
13.7
-
4-hydroxy-4-methyl-2-oxoglutarate
wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C
Pseudomonas putida
19.3
-
4-hydroxy-4-methyl-2-oxoglutarate
wild type protein, 1.0 mM Mg2+, pH 8.0, 25°C
Pseudomonas putida
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
additional information
-
broad optimum (log KCAT) pH 8 - pH 10.5
Pseudomonas putida
pH Range
pH Minimum
pH Maximum
Commentary
Organism
6
10.5
-
Pseudomonas putida
Ki Value [mM]
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.0017
-
oxalate
1 mM MnCl, pH 8.0, 25°C
Pseudomonas putida
0.005
-
oxalate
1 mM MgCl, pH 8.0, 25°C
Pseudomonas putida
Cloned(Commentary) (protein specific)
Commentary
Organism
expressed in Escherichia coli BL21(lambdaDE3)
Pseudomonas putida
Crystallization (Commentary) (protein specific)
Crystallization
Organism
hanging drop method, PEG 4000
Pseudomonas putida
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
R123A
no activity
Pseudomonas putida
R123K
retains some activity
Pseudomonas putida
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
oxalate
competitive inhibitor
Pseudomonas putida
Ki Value [mM] (protein specific)
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.0017
-
oxalate
1 mM MnCl, pH 8.0, 25°C
Pseudomonas putida
0.005
-
oxalate
1 mM MgCl, pH 8.0, 25°C
Pseudomonas putida
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.022
-
4-hydroxy-4-methyl-2-oxoglutarate
wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C
Pseudomonas putida
0.036
-
oxaloacetate
wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C; substrate inhibition constante 0.25 mM
Pseudomonas putida
0.071
-
2-hydroxy-4-oxopentanedioic acid
wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C
Pseudomonas putida
0.15
-
2-hydroxy-4-oxopentanedioic acid
wild type protein, 1.0 mM Mg2+, pH 8.0, 25°C
Pseudomonas putida
0.26
-
4-hydroxy-4-methyl-2-oxoglutarate
wild type protein, 1.0 mM Mg2+, pH 8.0, 25°C
Pseudomonas putida
0.3
-
oxaloacetate
wild type protein, 1.0 mM Mg2+, pH 8.0, 25°C; substrate inhibition constante 0.37 mM
Pseudomonas putida
5.5
-
4-hydroxy-4-methyl-2-oxoglutarate
R123K mutant protein, pH 8.0, 25°C
Pseudomonas putida
8.8
-
4-hydroxy-2-oxopentanoate
wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C
Pseudomonas putida
25
-
4-hydroxy-2-oxopentanoate
wild type protein, 1.0 mM Mg2+, pH 8.0, 25°C
Pseudomonas putida
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Mg2+
Mn2+: 90.3% activity, Co2+: 64.7% activity, Zn2+: 49.6% activity
Pseudomonas putida
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
26000
-
6 * 26000, crystal structure, gel filtration and SDS-PAGE
Pseudomonas putida
Purification (Commentary) (protein specific)
Commentary
Organism
ion exchange chromatography, hydrophobic interaction chromatography, gel filtration
Pseudomonas putida
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
37.7
-
wild type protein, pH 8.0, 25°C
Pseudomonas putida
Storage Stability (protein specific)
Storage Stability
Organism
-80°C, 20 mM sodium HEPES buffer, pH7.5, stable for 2 months
Pseudomonas putida
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2-hydroxy-4-oxopentanedioic acid
-
715556
Pseudomonas putida
?
-
-
-
?
4-hydroxy-2-oxopentanoate
-
715556
Pseudomonas putida
?
-
-
-
?
4-hydroxy-4-methyl-2-oxoglutarate
-
715556
Pseudomonas putida
2 pyruvate
-
-
-
r
oxaloacetate
-
715556
Pseudomonas putida
?
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
homohexamer
6 * 26000, crystal structure, gel filtration and SDS-PAGE
Pseudomonas putida
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.048
-
4-hydroxy-2-oxopentanoate
wild type protein, 1.0 mM Mg2+, pH 8.0, 25°C
Pseudomonas putida
0.058
-
2-hydroxy-4-oxopentanedioic acid
wild type protein, 1.0 mM Mg2+, pH 8.0, 25°C
Pseudomonas putida
0.099
-
4-hydroxy-4-methyl-2-oxoglutarate
R123K mutant protein, pH 8.0, 25°C
Pseudomonas putida
0.19
-
4-hydroxy-2-oxopentanoate
wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C
Pseudomonas putida
0.25
-
2-hydroxy-4-oxopentanedioic acid
wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C
Pseudomonas putida
1.52
-
oxaloacetate
wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C
Pseudomonas putida
1.85
-
oxaloacetate
wild type protein, 1.0 mM Mg2+, pH 8.0, 25°C
Pseudomonas putida
13.7
-
4-hydroxy-4-methyl-2-oxoglutarate
wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C
Pseudomonas putida
19.3
-
4-hydroxy-4-methyl-2-oxoglutarate
wild type protein, 1.0 mM Mg2+, pH 8.0, 25°C
Pseudomonas putida
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
additional information
-
broad optimum (log KCAT) pH 8 - pH 10.5
Pseudomonas putida
pH Range (protein specific)
pH Minimum
pH Maximum
Commentary
Organism
6
10.5
-
Pseudomonas putida
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.0019
-
4-hydroxy-2-oxopentanoate
wild type protein, 1.0 mM Mg2+, pH 8.0, 25°C
Pseudomonas putida
0.022
-
4-hydroxy-2-oxopentanoate
wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C
Pseudomonas putida
0.38
-
2-hydroxy-4-oxopentanedioic acid
wild type protein, 1.0 mM Mg2+, pH 8.0, 25°C
Pseudomonas putida
3.5
-
2-hydroxy-4-oxopentanedioic acid
wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C
Pseudomonas putida
6.2
-
oxaloacetate
wild type protein, 1.0 mM Mg2+, pH 8.0, 25°C
Pseudomonas putida
74
-
4-hydroxy-4-methyl-2-oxoglutarate
wild type protein, 1.0 mM Mg2+, pH 8.0, 25°C
Pseudomonas putida
420
-
oxaloacetate
wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C
Pseudomonas putida
620
-
4-hydroxy-4-methyl-2-oxoglutarate
wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C
Pseudomonas putida
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.0019
-
4-hydroxy-2-oxopentanoate
wild type protein, 1.0 mM Mg2+, pH 8.0, 25°C
Pseudomonas putida
0.022
-
4-hydroxy-2-oxopentanoate
wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C
Pseudomonas putida
0.38
-
2-hydroxy-4-oxopentanedioic acid
wild type protein, 1.0 mM Mg2+, pH 8.0, 25°C
Pseudomonas putida
3.5
-
2-hydroxy-4-oxopentanedioic acid
wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C
Pseudomonas putida
6.2
-
oxaloacetate
wild type protein, 1.0 mM Mg2+, pH 8.0, 25°C
Pseudomonas putida
74
-
4-hydroxy-4-methyl-2-oxoglutarate
wild type protein, 1.0 mM Mg2+, pH 8.0, 25°C
Pseudomonas putida
420
-
oxaloacetate
wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C
Pseudomonas putida
620
-
4-hydroxy-4-methyl-2-oxoglutarate
wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C
Pseudomonas putida
Other publictions for EC 4.1.3.17
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
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)
749095
Mazurkewich
Investigation into the mode o ...
Pseudomonas putida
PLoS ONE
11
e0164556
2016
2
-
-
-
8
-
-
22
-
-
-
2
-
2
-
-
-
-
-
-
-
-
3
-
-
-
-
21
-
-
-
-
-
-
-
2
-
-
-
-
8
-
-
-
-
22
-
-
-
2
-
-
-
-
-
-
-
-
3
-
-
-
-
21
-
-
-
-
-
-
-
-
21
21
727049
Mazurkewich
Biochemical and structural ana ...
Pseudomonas putida, Saccharomyces cerevisiae, Thermus thermophilus, Thermus thermophilus HB8 / ATCC 27634 / DSM 579
Biochemistry
53
542-553
2014
-
-
1
-
1
-
3
4
-
8
-
2
-
98
-
-
-
-
-
-
-
-
8
1
3
-
-
4
3
-
-
-
2
-
-
-
-
1
-
-
1
-
-
3
2
4
-
8
-
2
-
-
-
-
-
-
-
-
8
1
3
-
-
4
3
-
-
-
-
5
5
-
4
4
715556
Wang
Structural and kinetic charact ...
Pseudomonas putida
J. Biol. Chem.
285
36608-36615
2010
-
-
1
1
2
-
1
9
-
1
1
-
-
2
-
-
1
-
-
-
1
1
4
1
-
-
-
9
1
1
-
-
2
-
-
-
-
1
-
1
2
-
-
1
2
9
-
1
1
-
-
-
-
1
-
-
1
1
4
1
-
-
-
9
1
1
-
-
-
-
-
-
8
8
664301
Griffiths
A bacterial selection for the ...
Escherichia coli
Bioorg. Med. Chem.
12
4067-4074
2004
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
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-
-
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-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
650628
Maruyama
Cloning, sequencing, and expre ...
Pseudomonas straminea, Pseudomonas straminea NGJ1
Biosci. Biotechnol. Biochem.
65
2701-2709
2001
-
-
1
-
-
-
-
6
1
3
3
3
-
6
-
-
1
-
-
-
1
-
12
1
-
-
-
-
1
-
-
-
-
2
-
-
-
1
-
-
-
-
-
-
-
6
1
3
3
3
-
-
-
1
-
-
1
-
12
1
-
-
-
-
1
-
-
2
-
-
-
-
-
-
33194
Maruyama
Chemical modification of Pseud ...
Pseudomonas straminea
J. Biochem.
110
976-981
1991
-
-
-
-
-
-
1
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
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-
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-
-
-
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-
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-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
33192
Maruyama
Activation of Pseudomonas ochr ...
Pseudomonas straminea
J. Biochem.
10
334-340
1990
1
-
-
-
-
-
3
-
-
1
-
1
-
2
-
-
-
-
-
-
1
-
6
-
-
-
1
-
-
-
1
-
-
-
-
1
-
-
-
-
-
-
-
3
-
-
-
1
-
1
-
-
-
-
-
-
1
-
6
-
-
-
1
-
-
-
1
-
-
-
-
-
-
-
33193
Maruyama
Purification and properties of ...
Pseudomonas straminea
J. Biochem.
108
327-333
1990
-
-
-
-
-
-
7
6
-
5
2
-
-
2
-
-
1
-
-
-
-
-
8
1
-
-
-
-
5
-
-
-
-
-
-
-
-
-
-
-
-
-
-
7
-
6
-
5
2
-
-
-
-
1
-
-
-
-
8
1
-
-
-
-
5
-
-
-
-
-
-
-
-
-
33191
Dagley
4-Hydroxy-4-methyl-2-ketogluta ...
Pseudomonas putida
Methods Enzymol.
90
272-276
1982
-
-
-
-
-
-
-
2
-
3
1
-
-
2
-
-
1
-
-
-
1
-
2
-
-
-
-
-
7
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
3
1
-
-
-
-
1
-
-
1
-
2
-
-
-
-
-
7
-
-
-
-
-
-
-
-
-
33189
Schuld Ritter
Absolute configuration of a me ...
Comamonas testosteroni
J. Bacteriol.
113
1064-1065
1973
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
1
-
-
-
-
-
-
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4971
Wood
-
2-Keto-3-deoxy-6-phosphoglucon ...
Arachis hypogaea
The Enzymes, 3rd Ed. (Boyer, P. D. , ed. )
7
281-302
1972
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2
1
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33188
Tack
Purification and properties of ...
Pseudomonas putida
J. Biol. Chem.
247
6444-6449
1972
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2
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33187
Shannon
gamma-Methyl-gamma-hydroxy-alp ...
Arachis hypogaea
J. Biol. Chem.
237
3342-3347
1962
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