Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli | [Clostridium] symbiosum |
Protein Variants | Comment | Organism |
---|---|---|
W243F | decreased activity compared to the wild type enzyme, more thermostable than the wild type enzyme | [Clostridium] symbiosum |
W310F | more thermostable than the wild type enzyme | [Clostridium] symbiosum |
W393F | increased activity compared to the wild type enzyme, more thermostable than the wild type enzyme | [Clostridium] symbiosum |
W449F | decreased activity compared to the wild type enzyme, more thermostable than the wild type enzyme | [Clostridium] symbiosum |
W64F | 65% wild type enzyme activity, less thermostable than the wild type enzyme | [Clostridium] symbiosum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.125 | - |
NAD+ | wild type enzyme, in 0.1 M potassium phosphate buffer, at pH 7.0 and 25°C | [Clostridium] symbiosum | |
0.138 | - |
NAD+ | mutant enzyme W393F, in 0.1 M potassium phosphate buffer, at pH 7.0 and 25°C | [Clostridium] symbiosum | |
0.22 | - |
NAD+ | mutant enzyme W449F, in 0.1 M potassium phosphate buffer, at pH 7.0 and 25°C | [Clostridium] symbiosum | |
0.23 | - |
NAD+ | mutant enzyme W64F, in 0.1 M potassium phosphate buffer, at pH 7.0 and 25°C | [Clostridium] symbiosum | |
0.35 | - |
NAD+ | mutant enzyme W310F, in 0.1 M potassium phosphate buffer, at pH 7.0 and 25°C | [Clostridium] symbiosum | |
0.4 | - |
NAD+ | mutant enzyme W243F, in 0.1 M potassium phosphate buffer, at pH 7.0 and 25°C | [Clostridium] symbiosum | |
3.25 | - |
L-glutamate | mutant enzyme W393F, in 0.1 M potassium phosphate buffer, at pH 7.0 and 25°C | [Clostridium] symbiosum | |
3.25 | - |
L-glutamate | wild type enzyme, in 0.1 M potassium phosphate buffer, at pH 7.0 and 25°C | [Clostridium] symbiosum | |
3.3 | - |
L-glutamate | mutant enzyme W449F, in 0.1 M potassium phosphate buffer, at pH 7.0 and 25°C | [Clostridium] symbiosum | |
3.41 | - |
L-glutamate | mutant enzyme W64F, in 0.1 M potassium phosphate buffer, at pH 7.0 and 25°C | [Clostridium] symbiosum | |
3.7 | 5 | L-glutamate | mutant enzyme W310F, in 0.1 M potassium phosphate buffer, at pH 7.0 and 25°C | [Clostridium] symbiosum | |
18.2 | - |
L-glutamate | mutant enzyme W243F, in 0.1 M potassium phosphate buffer, at pH 7.0 and 25°C | [Clostridium] symbiosum |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
[Clostridium] symbiosum | - |
- |
- |
Purification (Comment) | Organism |
---|---|
Remazol-Red column chromatography | [Clostridium] symbiosum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-glutamate + H2O + NAD+ | - |
[Clostridium] symbiosum | 2-oxoglutarate + NH3 + NADH + H+ | - |
? |
Subunits | Comment | Organism |
---|---|---|
hexamer | x-ray crystallography | [Clostridium] symbiosum |
Synonyms | Comment | Organism |
---|---|---|
GDH | - |
[Clostridium] symbiosum |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
the wild-type GDH retains only approximately 48% of starting activity after incubation at 50°C for 30 min in potassium phosphate (pH 7.0), mutant enzymes W243F and W449F show slightly enhanced stability, retaining approximately 5% more activity over the 30 min period, mutants W310F and W393F show greater stabilization retaining 78% and 82% of their activities, respectively, W64F retains only 37% activity after 30 min | [Clostridium] symbiosum |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
7 | - |
the wild type enzyme shows pH-dependent inactivation and conformational change and loses 91% of its activity over a few min on transfer from pH 7.0 to pH 8.8 | [Clostridium] symbiosum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
[Clostridium] symbiosum |