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Literature summary for 1.4.1.4 extracted from
- Biochemical characterization of two glutamate dehydrogenases with different cofactor specificities from a hyperthermophilic archaeon Pyrobaculum calidifontis (2013), Extremophiles, 17, 379-389.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene pcal_1606, DNA and amino acid sequence determination and analysis, phylogenetic analysis, expression of N-terminally Met-Ala-Ser-tagged and C-terminally His6-tagged enzyme in Eschericia coli strain Rosetta(DE3) | Pyrobaculum calidifontis |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.017 | - |
NADPH | pH 9.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis |  |
| 0.035 | - |
NADP+ | pH 10.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis | |
| 1.7 | - |
2-oxoglutarate | pH 9.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis | |
| 2.2 | - |
NH3 | pH 9.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis | |
| 3.4 | - |
L-glutamate | pH 10.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 47700 | - |
6 * 47700, tagged recombinant enzyme, SDS-PAGE | Pyrobaculum calidifontis |
| 270000 | - |
gel filtration, tagged recombinant enzyme | Pyrobaculum calidifontis |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| L-glutamate + H2O + NADP+ | Pyrobaculum calidifontis | - |
2-oxoglutarate + NH3 + NADPH + H+ | - |
r | |
| L-glutamate + H2O + NADP+ | Pyrobaculum calidifontis JCM 11548 | - |
2-oxoglutarate + NH3 + NADPH + H+ | - |
r | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Pyrobaculum calidifontis | A3MWK6 | genes pcal_1031 and pcal_1606 | - |
| Pyrobaculum calidifontis JCM 11548 | A3MWK6 | genes pcal_1031 and pcal_1606 | - |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant N-terminally Met-Ala-Ser-tagged and C-terminally His6-tagged enzyme from Eschericia coli strain Rosetta(DE3) by affinity chromatography and gel filtration | Pyrobaculum calidifontis |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| L-glutamate + H2O + NADP+ | - |
Pyrobaculum calidifontis | 2-oxoglutarate + NH3 + NADPH + H+ | - |
r | |
| L-glutamate + H2O + NADP+ | - |
Pyrobaculum calidifontis JCM 11548 | 2-oxoglutarate + NH3 + NADPH + H+ | - |
r | |
| additional information | the enzyme also shows low activity with NAD+/NADH, 17% and 7% of the activity with NADP+ and NADPH, respectively. The enzyme also shows low activity with L-norvaline as substrates for oxidative deamination, and with 2-oxovalerate and 2-oxobutyrate for reducive amination, substrate specificity, overview. No activity with L-glutamine, L-alanine, L-aspartate, L-cysteine, L-serine, L-lysine, L-phenylalanine, and L-tryptophan, or with 2-oxoisocaproate, 2-oxocaproate, and pyruvate | Pyrobaculum calidifontis | ? | - |
? | |
| additional information | the enzyme also shows low activity with NAD+/NADH, 17% and 7% of the activity with NADP+ and NADPH, respectively. The enzyme also shows low activity with L-norvaline as substrates for oxidative deamination, and with 2-oxovalerate and 2-oxobutyrate for reducive amination, substrate specificity, overview. No activity with L-glutamine, L-alanine, L-aspartate, L-cysteine, L-serine, L-lysine, L-phenylalanine, and L-tryptophan, or with 2-oxoisocaproate, 2-oxocaproate, and pyruvate | Pyrobaculum calidifontis JCM 11548 | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| hexamer | 6 * 47700, tagged recombinant enzyme, SDS-PAGE | Pyrobaculum calidifontis |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| L-glutamate dehydrogenase | - |
Pyrobaculum calidifontis |
| Pcal_1606 | - |
Pyrobaculum calidifontis |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 50 | - |
assay at | Pyrobaculum calidifontis |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 90 | - |
Pcal_1606 retains full activity after incubation for 10 min at temperatures up to 90°C | Pyrobaculum calidifontis |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 9.9 | - |
NADP+ | pH 10.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis | |
| 13 | - |
L-glutamate | pH 10.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis | |
| 92 | - |
NADPH | pH 9.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis | |
| 95 | - |
2-oxoglutarate | pH 9.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis | |
| 100 | - |
NH3 | pH 9.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 9 | - |
reductive deamination | Pyrobaculum calidifontis |
| 9.5 | - |
oxidative deamination | Pyrobaculum calidifontis |
pH Stability
| pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|
| 4.5 | 9.5 | Pcal_1606 retains more than 80% of its activity after incubation for 30 min at pH 4.5-9.5 at 50°C | Pyrobaculum calidifontis |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| additional information | low activity with NAD+/NADH | Pyrobaculum calidifontis | |
| NADP+ | - |
Pyrobaculum calidifontis | |
| NADPH | - |
Pyrobaculum calidifontis | |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 3.8 | - |
L-glutamate | pH 10.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis | |
| 45 | - |
NH3 | pH 9.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis | |
| 56 | - |
2-oxoglutarate | pH 9.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis | |
| 280 | - |
NADP+ | pH 10.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis | |
| 5400 | - |
NADPH | pH 9.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis | |
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