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Literature summary for 1.4.1.3 extracted from

  • Maulik, P.; Ghosh, S.
    NADPH/NADH-dependent cold-labile glutamate dehydrogenase in Azospirillum brasilense. Purification and properties (1986), Eur. J. Biochem., 155, 595-602.
    View publication on PubMed

General Stability

General Stability Organism
glycerol, 20% v/v, protects against cold inactivation Azospirillum brasilense

Inhibitors

Inhibitors Comment Organism Structure
alanine weak inhibition at pH 8.5, strong inhibition at pH 10.0 Azospirillum brasilense
glutamate weak inhibition at pH 8.5, strong inhibition at pH 10.0 Azospirillum brasilense
histidine weak inhibition at pH 8.5, strong inhibition at pH 10.0 Azospirillum brasilense
lysine weak inhibition at pH 8.5, strong inhibition at pH 10.0 Azospirillum brasilense

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.004
-
NADP+ NADP+-dependent activity Azospirillum brasilense
0.013
-
NADPH NADPH-dependent activity Azospirillum brasilense
0.25
-
2-oxoglutarate NADPH-dependent activity Azospirillum brasilense
0.38
-
NH3 biphasic Lineweaver-Burk plot suggests 2 Km values Azospirillum brasilense
0.5
-
NADH NADH-dependent activity Azospirillum brasilense
5
-
2-oxoglutarate NADH-dependent activity Azospirillum brasilense
10
-
L-glutamate NADP+-dependent activity Azospirillum brasilense
66
-
NH3 NADH-dependent activity Azospirillum brasilense
100
-
NH3 biphasic Lineweaver-Burk plot suggests 2 Km values Azospirillum brasilense

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
48000
-
6 * 48000, SDS-PAGE Azospirillum brasilense
285000
-
gel filtration Azospirillum brasilense

Organism

Organism UniProt Comment Textmining
Azospirillum brasilense
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Azospirillum brasilense

Specific Activity [micromol/min/mg]

Specific Activity Minimum [┬Ámol/min/mg] Specific Activity Maximum [┬Ámol/min/mg] Comment Organism
41.9
-
-
Azospirillum brasilense

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-oxoglutarate + NH3 + NADH + H+
-
Azospirillum brasilense L-glutamate + H2O + NAD+
-
r
2-oxoglutarate + NH3 + NADH + H+ NH4Cl used in enzyme assay Azospirillum brasilense L-glutamate + H2O + NAD+
-
r
2-oxoglutarate + NH3 + NADPH + H+ NH4Cl used in enzyme assay Azospirillum brasilense L-glutamate + H2O + NADP+
-
r
L-glutamate + H2O + NAD(P)+ highly specific for 2-oxoglutarate and glutamate Azospirillum brasilense 2-oxoglutarate + NH3 + NAD(P)H
-
r

Subunits

Subunits Comment Organism
hexamer 6 * 48000, SDS-PAGE Azospirillum brasilense

Temperature Stability [┬░C]

Temperature Stability Minimum [┬░C] Temperature Stability Maximum [┬░C] Comment Organism
-
-
complete loss of activity after 5 h, 20% glycerol protects from inactivation Azospirillum brasilense
5
-
moderately stable above Azospirillum brasilense

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.33
-
L-glutamate cofactor NAD+, value below 200.0 Azospirillum brasilense
61
-
glutamate cofactor NADP+ Azospirillum brasilense
183
-
2-oxoglutarate cofactor NADH Azospirillum brasilense
200
-
NADPH NADPH-dependent activity Azospirillum brasilense

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.4 10 NADPH + 2-oxoglutarate + NH3, potassium phosphate buffer Azospirillum brasilense
8.5 8.6 NADPH + NH3 + 2-oxoglutarate, Tris-HCl buffer Azospirillum brasilense

pH Stability

pH Stability pH Stability Maximum Comment Organism
7 9 room temperature, 20% v/v glycerol, stable, room temperature, 1% v/v glycerol, 25% loss of activity after 48 h Azospirillum brasilense

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.1
-
glutamate at pH 10.0 Azospirillum brasilense
3.7
-
lysine at pH 10.0 Azospirillum brasilense
4.1
-
glutamate at pH 8.5 Azospirillum brasilense
6.1
-
histidine at pH 10.0 Azospirillum brasilense
6.2
-
alanine at pH 10.0 Azospirillum brasilense
9.3
-
lysine at pH 8.5 Azospirillum brasilense
23.2
-
alanine at pH 8.5 Azospirillum brasilense