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Literature summary for 1.4.1.3 extracted from

  • Bolivar, J.M.; Cava, F.; Mateo, C.; Rocha-Martin, J.; Guisan, J.M.; Berenguer, J.; Fernandez-Lafuente, R.
    Immobilization-stabilization of a new recombinant glutamate dehydrogenase from Thermus thermophilus (2008), Appl. Microbiol. Biotechnol., 80, 49-58.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21DE3 cells Thermus thermophilus

General Stability

General Stability Organism
glyoxyl agarose-GDH immobilization is more stable than CNBr-agarose preparations or than the soluble enzyme, glyoxyl agarose derivative, prepared at 4┬░C for 15 min, immobilizes around 50% of the enzyme, but the enzyme retains almost full activity after the immobilization Thermus thermophilus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.02
-
NAD+ in situ activity in 50 mM Tris-HCl buffer and 2 mM EDTA, pH 8, at 37┬░C Thermus thermophilus
0.035
-
NADP+ in situ activity in 50 mM Tris-HCl buffer and 2 mM EDTA, pH 8, at 37┬░C Thermus thermophilus
0.04
-
NADH in situ activity in 50 mM Tris-HCl buffer and 2 mM EDTA, pH 8, at 37┬░C Thermus thermophilus
3
-
L-glutamate in situ activity in 50 mM Tris-HCl buffer and 2 mM EDTA, pH 8, at 37┬░C Thermus thermophilus
3.5
-
2-oxoglutarate in situ activity in 50 mM Tris-HCl buffer and 2 mM EDTA, pH 8, at 37┬░C Thermus thermophilus
10
-
NH3 in situ activity in 50 mM Tris-HCl buffer and 2 mM EDTA, pH 8, at 37┬░C Thermus thermophilus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
42000
-
SDS-PAGE Thermus thermophilus

Organism

Organism UniProt Comment Textmining
Thermus thermophilus
-
-
-
Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039
-
-
-

Purification (Commentary)

Purification (Comment) Organism
heat denaturation at 70┬░C and Ni-agarose affinity column chromatography Thermus thermophilus

Storage Stability

Storage Stability Organism
25┬░C, soluble enzyme in the presence of 25% v/v acetone at pH 7, 1 week, no loss of activity Thermus thermophilus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-glutamate + H2O + NAD(P)+
-
Thermus thermophilus 2-oxoglutarate + NH3 + NAD(P)H
-
?
L-glutamate + H2O + NAD(P)+
-
Thermus thermophilus HB27 / ATCC BAA-163 / DSM 7039 2-oxoglutarate + NH3 + NAD(P)H
-
?

Subunits

Subunits Comment Organism
trimer analytical ultracentrifugation Thermus thermophilus

Synonyms

Synonyms Comment Organism
dual-coenzyme specific glutamate dehydrogenase
-
Thermus thermophilus
GDH
-
Thermus thermophilus

Temperature Stability [┬░C]

Temperature Stability Minimum [┬░C] Temperature Stability Maximum [┬░C] Comment Organism
25 70 the thermostability of the enzyme at neutral pH is very high even at 70┬░C, but at acidic pH values, the dissociation of enzyme subunits produces the rapid enzyme inactivation even at 25┬░C, immobilized preparations, as well as the soluble enzyme, remain fully active after 24 h of incubation at 60┬░C and pH 7, the optimal glyoxyl agarose derivative obtained is fully stable at pH 4 and 25┬░C, retaining more than 90% of its activity after incubation at 45┬░C for 24 h at pH 4 and more than 75% of the activity after the same period at 50┬░C Thermus thermophilus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9
-
immobilized enzyme, using NAD+ as a cofactor Thermus thermophilus

pH Stability

pH Stability pH Stability Maximum Comment Organism
4 7 the immobilized enzyme is very stable at pH 4, the enzyme activity does not decrease after 12 h at 45┬░C at pH 4, immobilized preparations, as well as the soluble enzyme, remain fully active after 24 h of incubation at 60┬░C and pH 7 Thermus thermophilus

Cofactor

Cofactor Comment Organism Structure
NAD+ activity with NAD+ is 3-5fold higher than using NADP+ Thermus thermophilus
NADP+ lower activity with NADP+ compared to NAD+ Thermus thermophilus