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Literature summary for 4.1.3.B3 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

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 (substrate 4-hydroxy-4-methyl-2-oxoglutarate) Pseudomonas putida

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.03
-
4-carboxy-4-hydroxy-2-oxoadipate wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C Pseudomonas putida
0.066
-
4-carboxy-4-hydroxy-2-oxoadipate wild type protein, 1.0 mM Mg2+, pH 8.0, 25°C Pseudomonas putida

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+
-
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 A5W059
-
-

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.
4-carboxy-4-hydroxy-2-oxoadipate
-
Pseudomonas putida pyruvate + oxaloacetate
-
r

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
7.5
-
4-carboxy-4-hydroxy-2-oxoadipate wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C Pseudomonas putida
14.4
-
4-carboxy-4-hydroxy-2-oxoadipate wild type protein, 1.0 mM Mg2+, 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
220
-
4-carboxy-4-hydroxy-2-oxoadipate wild type protein, 1.0 mM Mg2+, pH 8.0, 25°C Pseudomonas putida
250
-
4-carboxy-4-hydroxy-2-oxoadipate wild type protein, 1.0 mM Mn2+, pH 8.0, 25°C Pseudomonas putida