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Literature summary for 4.1.3.17 extracted from

  • Wang, W.; Mazurkewich, S.; Kimber, M.S.; Seah, S.Y.
    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 (2010), J. Biol. Chem., 285, 36608-36615.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 4.1.3.17

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(lambdaDE3) Pseudomonas putida

Crystallization (Commentary)

Crystallization (Comment) Organism
hanging drop method, PEG 4000 Pseudomonas putida

Protein Variants

Protein Variants Comment Organism
R123A no activity Pseudomonas putida
R123K retains some activity Pseudomonas putida

Inhibitors

Inhibitors Comment Organism Structure
oxalate competitive inhibitor Pseudomonas putida

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment 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 Comment 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] Comment Organism
26000
-
 6 * 26000, crystal structure, gel filtration and SDS-PAGE Pseudomonas putida

Organism

Organism UniProt Comment Textmining
Pseudomonas putida
-
-
-

Purification (Commentary)

Purification (Comment) 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] Comment 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 Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-hydroxy-4-oxopentanedioic acid
-
 Pseudomonas putida ?
-
 ?
4-hydroxy-2-oxopentanoate
-
 Pseudomonas putida ?
-
 ?
4-hydroxy-4-methyl-2-oxoglutarate
-
 Pseudomonas putida 2 pyruvate
-
 r
oxaloacetate
-
 Pseudomonas putida ?
-
 ?

Subunits

Subunits Comment 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 Comment 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 Comment Organism
additional information
-
 broad optimum (log KCAT) pH 8 - pH 10.5 Pseudomonas putida

pH Range

pH Minimum pH Maximum Comment Organism
6 10.5
-
 Pseudomonas putida

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment 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

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment 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