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Literature summary for 1.4.1.2 extracted from

  • Wakamatsu, T.; Higashi, C.; Ohmori, T.; Doi, K.; Ohshima, T.
    Biochemical characterization of two glutamate dehydrogenases with different cofactor specificities from a hyperthermophilic archaeon Pyrobaculum calidifontis (2013), Extremophiles, 17, 379-389.
    View publication on PubMed
Search for more literature for 1.4.1.2

Cloned(Commentary)

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 [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 [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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Pyrobaculum calidifontis
-
 gene pcal_1031
-
Pyrobaculum calidifontis JCM 11548
-
 gene pcal_1031
-

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 + 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

Subunits Comment Organism
hexamer 6 * 48200, tagged recombinant enzyme, SDS-PAGE Pyrobaculum calidifontis

Synonyms

Synonyms Comment Organism
L-glutamate dehydrogenase
-
 Pyrobaculum calidifontis
Pcal_1031
-
 Pyrobaculum calidifontis

Temperature Optimum [°C]

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 [°C]

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 [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
9.5
-
 reductive deamination Pyrobaculum calidifontis
10.5
-
 oxidative deamination Pyrobaculum calidifontis

pH Stability

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

Cofactor Comment Organism Structure
NAD+
-
 Pyrobaculum calidifontis
NADH
-
 Pyrobaculum calidifontis

kcat/KM [mM/s]

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