Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Pseudomonas putida |
Crystallization (Comment) | Organism |
---|---|
in complex with the intermediate analog oxalate, Mg2+ and NADH, to 2.0 A resolution | Pseudomonas putida |
Protein Variants | Comment | Organism |
---|---|---|
D225A | mutation in metal ion-binding ligand, 20fold decrease in metal ion-binding affinity and a two-orders-of-magnitude decrease in catalysis | Pseudomonas putida |
D250A | mutation in metal ion-binding ligand, 10fold decrease in metal ion-binding affinity and a two-orders-of-magnitude decrease in catalysis | Pseudomonas putida |
R108L | 1.3% of wild-type catalytic rate | Pseudomonas putida |
R108Q | 8.7% of wild-type catalytic rate | Pseudomonas putida |
R98L | mutant is not expressed in stable form | Pseudomonas putida |
R98Q | 2.6% of wild-type catalytic rate | Pseudomonas putida |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.025 | - |
NAD+ | wild-type, pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
0.079 | - |
D-malate | wild-type, pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
0.093 | - |
D-malate | mutant R108Q, pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
0.096 | - |
NAD+ | mutant R98Q, pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
0.098 | - |
D-malate | mutant R98Q, pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
0.135 | - |
NAD+ | mutant R108Q, pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
0.18 | - |
D-malate | mutant R108L, pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
0.24 | - |
NAD+ | mutant R108L, pH 7.5, temperature not specified in the publication | Pseudomonas putida |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas putida | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant enzyme | Pseudomonas putida |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
tartrate + NAD+ = oxaloglycolate + NADH + H+ | lysyl amino group, Lys192 is the base responsible for the water-mediated proton abstraction from the C2 hydroxyl group of the substrate that begins the catalytic reaction, followed by hydride transfer to NAD. The hydroxyl group of Tyr141 functions as a general acid to protonate the enolate intermediate. Each substrate undergoes the initial hydride transfer | Pseudomonas putida |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-malate + NAD+ | - |
Pseudomonas putida | pyruvate + CO2 + NADH + H+ | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.015 | - |
NAD+ | mutant D225A, pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
0.06 | - |
NAD+ | mutant R108Q, pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
0.087 | - |
NAD+ | mutant R108L, pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
0.088 | - |
NAD+ | mutant D250A, pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
0.175 | - |
NAD+ | mutant R98Q, pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
6.5 | - |
NAD+ | wild-type, pH 7.5, temperature not specified in the publication | Pseudomonas putida |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.36 | - |
NAD+ | mutant R108L, pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
0.48 | - |
D-malate | mutant R108L, pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
1.8 | - |
NAD+ | mutant R98Q, pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
1.8 | - |
D-malate | mutant R98Q, pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
4.3 | - |
NAD+ | mutant R108Q, pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
6.2 | - |
D-malate | mutant R108Q, pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
82 | - |
D-malate | wild-type, pH 7.5, temperature not specified in the publication | Pseudomonas putida | |
260 | - |
NAD+ | wild-type, pH 7.5, temperature not specified in the publication | Pseudomonas putida |