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Literature summary for 1.4.1.3 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)

Cloned (Comment) Organism
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]

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

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
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
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
284000
-
gel filtration Thermococcus kodakarensis

Organism

Organism UniProt Comment Textmining
Thermococcus kodakarensis O59650
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Thermococcus kodakarensis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [┬Ámol/min/mg] Specific Activity Maximum [┬Ámol/min/mg] Comment Organism
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)

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

Subunits

Subunits Comment Organism
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
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]

Temperature Optimum [┬░C] Temperature Optimum Maximum [┬░C] Comment Organism
70
-
assay at Thermococcus kodakarensis
80
-
oxidative deamination and reductive amination Thermococcus kodakarensis

Temperature Range [┬░C]

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

pH Optimum

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