Cloned (Comment) | Organism |
---|---|
gene pcal_1031, 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 Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.001 | - |
NADH | pH 9.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis | |
0.028 | - |
NAD+ | pH 10.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis | |
0.92 | - |
2-oxoglutarate | pH 9.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis | |
5.3 | - |
L-glutamate | pH 10.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis | |
9.8 | - |
NH3 | pH 9.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
48200 | - |
6 * 48200, tagged recombinant enzyme, SDS-PAGE | Pyrobaculum calidifontis |
286000 | - |
gel filtration, tagged recombinant enzyme | Pyrobaculum calidifontis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-glutamate + H2O + NAD+ | Pyrobaculum calidifontis | - |
2-oxoglutarate + NH3 + NADH + H+ | - |
r | |
L-glutamate + H2O + NAD+ | Pyrobaculum calidifontis JCM 11548 | - |
2-oxoglutarate + NH3 + NADH + H+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pyrobaculum calidifontis | - |
gene pcal_1031 | - |
Pyrobaculum calidifontis JCM 11548 | - |
gene pcal_1031 | - |
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 | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-glutamate + H2O + NAD+ | - |
Pyrobaculum calidifontis | 2-oxoglutarate + NH3 + NADH + H+ | - |
r | |
L-glutamate + H2O + NAD+ | - |
Pyrobaculum calidifontis JCM 11548 | 2-oxoglutarate + NH3 + NADH + H+ | - |
r | |
additional information | no or poor activity with NADP+/NADPH in both reaction directions. The enzyme also shows low activity with L-norvaline, L-2-aminobutyrate, L-valine, L-isoleucine, and L-leucine as substrates for oxidative deamination, and with 2-oxovalerate, 2-oxobutyrate, and 2-oxocaproate, for reducive amination, substrate specificity, overview. No activity with L-2-aminobutyrate, L-valine, L-isoleucine, L-leucine, L-glutamine, L-alanine, L-aspartate, L-cysteine, L-serine, L-lysine, L-phenylalanine, and L-tryptophan, or with 2-oxoisocaproate and pyruvate | Pyrobaculum calidifontis | ? | - |
? | |
additional information | no or poor activity with NADP+/NADPH in both reaction directions. The enzyme also shows low activity with L-norvaline, L-2-aminobutyrate, L-valine, L-isoleucine, and L-leucine as substrates for oxidative deamination, and with 2-oxovalerate, 2-oxobutyrate, and 2-oxocaproate, for reducive amination, substrate specificity, overview. No activity with L-2-aminobutyrate, L-valine, L-isoleucine, L-leucine, L-glutamine, L-alanine, L-aspartate, L-cysteine, L-serine, L-lysine, L-phenylalanine, and L-tryptophan, or with 2-oxoisocaproate and pyruvate | Pyrobaculum calidifontis JCM 11548 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
hexamer | 6 * 48200, tagged recombinant enzyme, SDS-PAGE | Pyrobaculum calidifontis |
Synonyms | Comment | Organism |
---|---|---|
L-glutamate dehydrogenase | - |
Pyrobaculum calidifontis |
Pcal_1031 | - |
Pyrobaculum calidifontis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
100 | - |
Pcal_1031 activity appreciably increases by incubating at temperatures up to 100°C | Pyrobaculum calidifontis |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
105 | - |
Pcal_1031retains full activity after incubation for 10 min at temperatures up to 105°C | Pyrobaculum calidifontis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
14 | - |
NAD+ | pH 10.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis | |
17 | - |
L-glutamate | pH 10.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis | |
58 | - |
NADH | pH 9.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis | |
58 | - |
NH3 | pH 9.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis | |
65 | - |
2-oxoglutarate | pH 9.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
9.5 | - |
reductive deamination | Pyrobaculum calidifontis |
10.5 | - |
oxidative deamination | Pyrobaculum calidifontis |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
4 | 10.5 | Pcal_1031 retains more than 80% of its activity after incubation for 30 min at pH 4.0-10.5 at 50°C | Pyrobaculum calidifontis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Pyrobaculum calidifontis | |
NADH | - |
Pyrobaculum calidifontis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3.2 | - |
L-glutamate | pH 10.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis | |
5.9 | - |
NH3 | pH 9.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis | |
71 | - |
2-oxoglutarate | pH 9.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis | |
500 | - |
NAD+ | pH 10.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis | |
5800 | - |
NADH | pH 9.5, 50°C, recombinant enzyme | Pyrobaculum calidifontis |