Cloned (Comment) | Organism |
---|---|
expression in a strain of Haloferax volcanii lacking dihydrolipoamide dehydrogenase activity | Haloferax volcanii |
Protein Variants | Comment | Organism |
---|---|---|
E423A | in 2 M KCl, the mutant is significantly less active than wild-type, decreased Km value for dihydrolipoamide. The mutant enzyme is not significantly activated by high salt concentrations. In the presence of 2 M KCI the thermal stability of the mutant enzyme is slightly higher than that of the wild-type enzyme | Haloferax volcanii |
E423D | wild-type and E423D mutant enzyme are much less active in the absence of KCl than in its presence. The mutant enzyme is inactivated at temperatures around 20°C lower than the wild-type | Haloferax volcanii |
E423Q | in 2 M KCl, the mutant is significantly less active than wild-type, whereas Km, differences are not significant. The mutant enzyme is not significantly activated by high salt concentrations. In the presence of 2 M KCI the thermal stability of the mutant enzyme is slightly higher than that of the wild-type enzyme | Haloferax volcanii |
E423S | in 2 M KCl, the mutant is significantly less active than wild-type, whereas Km, differences are not significant. The mutant enzyme is not significantly activated by high salt concentrations. In the presence of 2 M KCI the thermal stability of the mutant enzyme is slightly higher than that of the wild-type enzyme | Haloferax volcanii |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
dihydrolipoamide | - |
Haloferax volcanii |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0107 | - |
dihydrolipoamide | pH 7.0, 30°C, mutant enzyme E423A | Haloferax volcanii | |
0.0157 | - |
dihydrolipoamide | pH 7.0, 30°C, mutant enzyme E423D, assay in the absence of KCI | Haloferax volcanii | |
0.0177 | - |
dihydrolipoamide | pH 7.0, 30°C, mutant enzyme E423Q, assay in the absence of KCI | Haloferax volcanii | |
0.0228 | - |
dihydrolipoamide | pH 7.0, 30°C, mutant enzyme E423Q | Haloferax volcanii | |
0.023 | - |
dihydrolipoamide | pH 7.0, 30°C, mutant enzyme E423S | Haloferax volcanii | |
0.0243 | - |
dihydrolipoamide | pH 7.0, 30°C, mutant enzyme E423A, assay in the absence of KCI | Haloferax volcanii | |
0.0266 | - |
dihydrolipoamide | pH 7.0, 30°C, mutant enzyme E423D | Haloferax volcanii | |
0.0269 | - |
dihydrolipoamide | pH 7.0, 30°C, wild-type enzyme | Haloferax volcanii | |
0.0274 | - |
dihydrolipoamide | pH 7.0, 30°C, wild-type enzyme, assay in the absence of KCI | Haloferax volcanii | |
0.0364 | - |
dihydrolipoamide | pH 7.0, 30°C, mutant enzyme E423S, assay in the absence of KCI | Haloferax volcanii | |
0.156 | - |
NAD+ | pH 7.0, 30°C, wild-type enzyme | Haloferax volcanii | |
0.182 | - |
NAD+ | pH 7.0, 30°C, mutant enzyme E423A, assay in the absence of KCI | Haloferax volcanii | |
0.186 | - |
NAD+ | pH 7.0, 30°C, wild-type enzyme, assay in the absence of KCI | Haloferax volcanii | |
0.199 | - |
NAD+ | pH 7.0, 30°C, mutant enzyme E423D | Haloferax volcanii | |
0.203 | - |
NAD+ | pH 7.0, 30°C, mutant enzyme E423Q | Haloferax volcanii | |
0.211 | - |
NAD+ | pH 7.0, 30°C, mutant enzyme E423S | Haloferax volcanii | |
0.215 | - |
NAD+ | pH 7.0, 30°C, mutant enzyme E423A | Haloferax volcanii | |
0.288 | - |
NAD+ | pH 7.0, 30°C, mutant enzyme E423Q, assay in the absence of KCI | Haloferax volcanii | |
0.292 | - |
NAD+ | pH 7.0, 30°C, mutant enzyme E423D, assay in the absence of KCI | Haloferax volcanii | |
0.312 | - |
NAD+ | pH 7.0, 30°C, mutant enzyme E423S, assay in the absence of KCI | Haloferax volcanii |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Haloferax volcanii | Q04829 | - |
- |
Haloferax volcanii DSM 3757 | Q04829 | - |
- |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
3.6 | - |
pH 7.0, 30°C, mutant enzyme E423Q, assay in the absence of KCI | Haloferax volcanii |
4.6 | - |
pH 7.0, 30°C, mutant enzyme E423A, assay in the absence of KCI | Haloferax volcanii |
4.9 | - |
pH 7.0, 30°C, mutant enzyme E423D, assay in the absence of KCI | Haloferax volcanii |
5 | - |
pH 7.0, 30°C, mutant enzyme E423S, assay in the absence of KCI | Haloferax volcanii |
5.6 | - |
pH 7.0, 30°C, wild-type enzyme, assay in the absence of KCI | Haloferax volcanii |
5.8 | - |
pH 7.0, 30°C, mutant enzyme E423S, assay in the presence of 2 M KCI | Haloferax volcanii |
6.2 | - |
pH 7.0, 30°C, mutant enzyme E423A, assay in the presence of 2 M KCI | Haloferax volcanii |
6.4 | - |
pH 7.0, 30°C, mutant enzyme E423Q, assay in the presence of 2 M KCI | Haloferax volcanii |
16.2 | - |
pH 7.0, 30°C, mutant enzyme E423D, assay in the presence of 2 M KCI | Haloferax volcanii |
19.1 | - |
pH 7.0, 30°C, wild-type enzyme, assay in the presence of 2 M KCI | Haloferax volcanii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
dihydrolipoamide + NAD+ | - |
Haloferax volcanii | lipoamide + NADH | - |
? | |
dihydrolipoamide + NAD+ | - |
Haloferax volcanii DSM 3757 | lipoamide + NADH | - |
? |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Haloferax volcanii |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
additional information | - |
in thepresence of 2 M KCI, the thermal stability properties of mutant enzymes E423S, E423Q and E423A are comparable to those of wild-type, although the mutants appear to be slightly more stable. Mutant enzyme E423D however, is inactivated at temperatures around 20°C lower than for the wild-type. In the absence of salt, similar relative thermostabilities are observed, but both wild-type and mutant enzymes are inactivated at temperatures around 16-18°C lower than in the presence of salt | Haloferax volcanii |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Haloferax volcanii |
8.5 | - |
wild-type enzyme and mutant enzymes E423D, E423Q, E423S, and E423A | Haloferax volcanii |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.023 | - |
dihydrolipoamide | pH 7.0, 30°C | Haloferax volcanii |