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Literature summary extracted from

  • Abd Rahman, R.N.; Fujiwara, S.; Takagi, M.; Kanaya, S.; Imanaka, T.
    Effect of heat treatment on proper oligomeric structure formation of thermostable glutamate dehydrogenase from a hyperthermophilic archaeon (1997), Biochem. Biophys. Res. Commun., 241, 646-652.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.4.1.3 expression in Escherichia coli. The specific activity of the hexameric form of the recombinant enzyme is much lower than that of the natural enzyme. The structure of the hexameric form of the recombinant enzyme with low specific activity (Type I) is different from that of the natural enzyme with high specific activity (Type II). Upon heat treatment (80°C, 15 min), the Type I structure is effectively converted to Type II structure and the specific activity of the enzyme is increased by 2.6fold Thermococcus kodakarensis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.4.1.3 0.04
-
 NADPH pH 7.6, 70°C, native enzyme Thermococcus kodakarensis
1.4.1.3 0.05
-
 NADPH pH 7.6, 70°C, recombinent enzyme (heated) Thermococcus kodakarensis
1.4.1.3 0.1
-
 NADPH pH 7.6, 70°C, recombinent enzyme (unheated) Thermococcus kodakarensis
1.4.1.3 0.2
-
 2-oxoglutarate pH 7.6, 70°C, recombinent enzyme (unheated) Thermococcus kodakarensis
1.4.1.3 0.25
-
 2-oxoglutarate pH 7.6, 70°C, native enzyme Thermococcus kodakarensis
1.4.1.3 0.3
-
 2-oxoglutarate pH 7.6, 70°C, recombinent enzyme (heated) Thermococcus kodakarensis
1.4.1.3 2.5
-
 NH3 pH 7.6, 70°C, recombinent enzyme (unheated) Thermococcus kodakarensis
1.4.1.3 11.4
-
 NH3 pH 7.6, 70°C, recombinent enzyme (heated) Thermococcus kodakarensis
1.4.1.3 13
-
 NH3 pH 7.6, 70°C, native enzyme Thermococcus kodakarensis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.4.1.3 47040
-
 6 * 47040, calculated from sequence, the natural enzyme was purified only as a hexameric form, whereas the recombinant enzyme was purified as both monomeric and hexameric forms. Only the enzyme in a hexameric form is active. Upon heat treatment (70°C for 15 min), the inactive monomeric form of the recombinant enzyme is at least partially associated with the hexameric form Thermococcus kodakarensis
1.4.1.3 47300
-
 6 * 47300, SDS-PAGE, the natural enzyme was purified only as a hexameric form, whereas the recombinant enzyme was purified as both monomeric and hexameric forms. Only the enzyme in a hexameric form is active. Upon heat treatment (70°C for 15 min), the inactive monomeric form of the recombinant enzyme is at least partially associated with the hexameric form Thermococcus kodakarensis
1.4.1.3 284000
-
 gel filtration Thermococcus kodakarensis

Organism

EC Number Organism UniProt Comment Textmining
1.4.1.3 Thermococcus kodakarensis O59650
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.4.1.3
-
 Thermococcus kodakarensis

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.4.1.3 additional information
-
 the specific activity of the hexameric form of the recombinant enzyme is much lower than that of the natural enzyme. The structure of the hexameric form of the recombinant enzyme with low specific activity (Type I) is different from that of the natural enzyme with high specific activity (Type II). Upon heat treatment (80°C, 15 min), the Type I structure is effectively converted to Type II structure and the specific activity of the enzyme is increased by 2.6fold Thermococcus kodakarensis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.4.1.3 2-oxoglutarate + NH3 + NADPH + H+
-
 Thermococcus kodakarensis L-glutamate + H2O + NADP+
-
 ?

Subunits

EC Number Subunits Comment Organism
1.4.1.3 hexamer 6 * 47040, calculated from sequence, the natural enzyme was purified only as a hexameric form, whereas the recombinant enzyme was purified as both monomeric and hexameric forms. Only the enzyme in a hexameric form is active. Upon heat treatment (70°C for 15 min), the inactive monomeric form of the recombinant enzyme is at least partially associated with the hexameric form Thermococcus kodakarensis
1.4.1.3 hexamer 6 * 47300, SDS-PAGE, the natural enzyme was purified only as a hexameric form, whereas the recombinant enzyme was purified as both monomeric and hexameric forms. Only the enzyme in a hexameric form is active. Upon heat treatment (70°C for 15 min), the inactive monomeric form of the recombinant enzyme is at least partially associated with the hexameric form Thermococcus kodakarensis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.4.1.3 70
-
 assay at Thermococcus kodakarensis
1.4.1.3 80
-
 oxidative deamination and reductive amination Thermococcus kodakarensis

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.4.1.3 40 90 40°C: about 35% of maximal activity, 90°C: about 75% of maximal activity, reductive amination Thermococcus kodakarensis
1.4.1.3 40 90 40°C: about 60% of maximal activity, 90°C: about 85% of maximal activity, oxidative deamination Thermococcus kodakarensis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.4.1.3 7.6
-
 assay at Thermococcus kodakarensis