Cloned (Comment) | Organism |
---|---|
cloned with the Escherichia coli expression vector pET11a, large amounts of a soluble but inactive form of the enzyme are produced upon its induction | Haloarcula marismortui |
Protein Variants | Comment | Organism |
---|---|---|
R100Q | the mutant enzyme has considerably higher specificity for pyruvate than for oxaloacetate. Whereas the Km value for pyruvate is increased about 2fold, that for oxaloacetate increases 30fold. The R100Q mutant is not subjected to substrate inhibition, at least not at substrate concentrations up to 30 mM, and the highest kcat value is obtained at the lowest salt concentration used (0.15 M NaCl) | Haloarcula marismortui |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.7 | - |
oxaloacetate | pH 7.0, temperature not specified in the publication, 4 M NaCl, wild-type enzyme | Haloarcula marismortui | |
0.9 | - |
oxaloacetate | pH 7.0, temperature not specified in the publication, 2 M NaCl, wild-type enzyme | Haloarcula marismortui | |
2.2 | - |
pyruvate | pH 7.0, temperature not specified in the publication, 0.15 M NaCl, mutant enzyme R100Q | Haloarcula marismortui | |
2.2 | - |
pyruvate | pH 7.0, temperature not specified in the publication, 2 M NaCl, mutant enzyme R100Q | Haloarcula marismortui | |
2.9 | - |
pyruvate | pH 7.0, temperature not specified in the publication, 4 M NaCl, mutant enzyme R100Q | Haloarcula marismortui | |
21.8 | - |
oxaloacetate | pH 7.0, temperature not specified in the publication, 4 M NaCl, mutant enzyme R100Q | Haloarcula marismortui | |
29.4 | - |
oxaloacetate | pH 7.0, temperature not specified in the publication, 2 M NaCl, mutant enzyme R100Q | Haloarcula marismortui |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
NaCl | activation of the enzyme was obtained by increasing the salt concentration to 3 M NaCl | Haloarcula marismortui |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
32638 | - |
x * 32638, calculated from sequence | Haloarcula marismortui |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Haloarcula marismortui | Q07841 | - |
- |
Haloarcula marismortui DSM 3752 | Q07841 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant enzyme | Haloarcula marismortui |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
oxaloacetate + NADH + H+ | - |
Haloarcula marismortui | (S)-malate + NAD+ | - |
? | |
oxaloacetate + NADH + H+ | - |
Haloarcula marismortui DSM 3752 | (S)-malate + NAD+ | - |
? | |
pyruvate + NADH + H+ | wild-type enzyme shows no activity with pyruvate. Mutant enzyme R100Q has considerably higher specificity for pyruvate than for oxaloacetate | Haloarcula marismortui | (S)-lactate + NAD+ | - |
? | |
pyruvate + NADH + H+ | wild-type enzyme shows no activity with pyruvate. Mutant enzyme R100Q has considerably higher specificity for pyruvate than for oxaloacetate | Haloarcula marismortui DSM 3752 | (S)-lactate + NAD+ | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 32638, calculated from sequence | Haloarcula marismortui |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.3 | - |
oxaloacetate | pH 7.0, temperature not specified in the publication, 4 M NaCl, mutant enzyme R100Q | Haloarcula marismortui | |
2 | - |
oxaloacetate | pH 7.0, temperature not specified in the publication, 2 M NaCl, mutant enzyme R100Q | Haloarcula marismortui | |
2.1 | - |
pyruvate | pH 7.0, temperature not specified in the publication, 2 M NaCl, mutant enzyme R100Q | Haloarcula marismortui | |
3.5 | - |
pyruvate | pH 7.0, temperature not specified in the publication, 4 M NaCl, mutant enzyme R100Q | Haloarcula marismortui | |
6 | 9 | oxaloacetate | pH 7.0, temperature not specified in the publication, 4 M NaCl, wild-type enzyme | Haloarcula marismortui | |
12.3 | - |
pyruvate | pH 7.0, temperature not specified in the publication, 0.15 M NaCl, mutant enzyme R100Q | Haloarcula marismortui | |
197 | - |
oxaloacetate | pH 7.0, temperature not specified in the publication, 2 M NaCl, wild-type enzyme | Haloarcula marismortui |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Haloarcula marismortui |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | - |
Haloarcula marismortui |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0014 | - |
oxaloacetate | pH 7.0, temperature not specified in the publication, 4 M NaCl, mutant enzyme R100Q | Haloarcula marismortui | |
0.007 | - |
oxaloacetate | pH 7.0, temperature not specified in the publication, 2 M NaCl, mutant enzyme R100Q | Haloarcula marismortui | |
1 | - |
pyruvate | pH 7.0, temperature not specified in the publication, 2 M NaCl, mutant enzyme R100Q | Haloarcula marismortui | |
1.2 | - |
pyruvate | pH 7.0, temperature not specified in the publication, 4 M NaCl, mutant enzyme R100Q | Haloarcula marismortui | |
5.6 | - |
pyruvate | pH 7.0, temperature not specified in the publication, 0.15 M NaCl, mutant enzyme R100Q | Haloarcula marismortui | |
100 | - |
oxaloacetate | pH 7.0, temperature not specified in the publication, 4 M NaCl, wild-type enzyme | Haloarcula marismortui | |
200 | - |
oxaloacetate | pH 7.0, temperature not specified in the publication, 2 M NaCl, wild-type enzyme | Haloarcula marismortui |