Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21DE3 cells | Thermus thermophilus |
General Stability | Organism |
---|---|
glyoxyl agarose-GDH immobilization is more stable than CNBr-agarose preparations or than the soluble enzyme, glyoxyl agarose derivative, prepared at 4°C for 15 min, immobilizes around 50% of the enzyme, but the enzyme retains almost full activity after the immobilization | Thermus thermophilus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.02 | - |
NAD+ | in situ activity in 50 mM Tris-HCl buffer and 2 mM EDTA, pH 8, at 37°C | Thermus thermophilus | |
0.035 | - |
NADP+ | in situ activity in 50 mM Tris-HCl buffer and 2 mM EDTA, pH 8, at 37°C | Thermus thermophilus | |
0.04 | - |
NADH | in situ activity in 50 mM Tris-HCl buffer and 2 mM EDTA, pH 8, at 37°C | Thermus thermophilus | |
3 | - |
L-glutamate | in situ activity in 50 mM Tris-HCl buffer and 2 mM EDTA, pH 8, at 37°C | Thermus thermophilus | |
3.5 | - |
2-oxoglutarate | in situ activity in 50 mM Tris-HCl buffer and 2 mM EDTA, pH 8, at 37°C | Thermus thermophilus | |
10 | - |
NH3 | in situ activity in 50 mM Tris-HCl buffer and 2 mM EDTA, pH 8, at 37°C | Thermus thermophilus |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
42000 | - |
SDS-PAGE | Thermus thermophilus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermus thermophilus | - |
- |
- |
Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
heat denaturation at 70°C and Ni-agarose affinity column chromatography | Thermus thermophilus |
Storage Stability | Organism |
---|---|
25°C, soluble enzyme in the presence of 25% v/v acetone at pH 7, 1 week, no loss of activity | Thermus thermophilus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-glutamate + H2O + NAD(P)+ | - |
Thermus thermophilus | 2-oxoglutarate + NH3 + NAD(P)H | - |
? | |
L-glutamate + H2O + NAD(P)+ | - |
Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039 | 2-oxoglutarate + NH3 + NAD(P)H | - |
? |
Subunits | Comment | Organism |
---|---|---|
trimer | analytical ultracentrifugation | Thermus thermophilus |
Synonyms | Comment | Organism |
---|---|---|
dual-coenzyme specific glutamate dehydrogenase | - |
Thermus thermophilus |
GDH | - |
Thermus thermophilus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | 70 | the thermostability of the enzyme at neutral pH is very high even at 70°C, but at acidic pH values, the dissociation of enzyme subunits produces the rapid enzyme inactivation even at 25°C, immobilized preparations, as well as the soluble enzyme, remain fully active after 24 h of incubation at 60°C and pH 7, the optimal glyoxyl agarose derivative obtained is fully stable at pH 4 and 25°C, retaining more than 90% of its activity after incubation at 45°C for 24 h at pH 4 and more than 75% of the activity after the same period at 50°C | Thermus thermophilus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9 | - |
immobilized enzyme, using NAD+ as a cofactor | Thermus thermophilus |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
4 | 7 | the immobilized enzyme is very stable at pH 4, the enzyme activity does not decrease after 12 h at 45°C at pH 4, immobilized preparations, as well as the soluble enzyme, remain fully active after 24 h of incubation at 60°C and pH 7 | Thermus thermophilus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | activity with NAD+ is 3-5fold higher than using NADP+ | Thermus thermophilus | |
NADP+ | lower activity with NADP+ compared to NAD+ | Thermus thermophilus |