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Literature summary for 1.4.1.13 extracted from

  • Adachi, K.; Suzuki, I.
    Purification and properties of glutamate synthase from Thiobacillus thioparus (1977), J. Bacteriol., 129, 1173-1182.
    View publication on PubMedView publication on EuropePMC

General Stability

General Stability Organism
2-mercaptoethanol, stabilizes Thiobacillus thioparus
DTT, 1 mM, stabilizes at 4°C Thiobacillus thioparus
EDTA, stabilizes Thiobacillus thioparus

Inhibitors

Inhibitors Comment Organism Structure
2,2'-bipyridyl 100% inhibition at 3 mM Thiobacillus thioparus
Atebrin 100% inhibition at 1 mM Thiobacillus thioparus
ATP 33% inhibition at 20 mM Thiobacillus thioparus
D-glutamate
-
Thiobacillus thioparus
iodoacetate 27% inhibition at 25 mM Thiobacillus thioparus
isocitrate 32% inhibition at 20 mM Thiobacillus thioparus
KCN 89% inhibition at 20 mM Thiobacillus thioparus
L-glutamate more than 40% inhibition at 50 mM Thiobacillus thioparus
L-histidine more than 40% inhibition at 50 mM Thiobacillus thioparus
L-methionine more than 40% inhibition at 50 mM Thiobacillus thioparus
L-serine more than 40% inhibition at 10 mM and 50 mM Thiobacillus thioparus
additional information a combination of L-serine, L-methionine, L-alanine, L-glycine, L-histidine and L-asparagine inhibits 73% of the acitivity at 7.5 mM each Thiobacillus thioparus
N-ethylmaleimide 83% inhibition at 10 mM Thiobacillus thioparus
NADP+ 59% inhibition at 1 mM Thiobacillus thioparus
NaN3 100% inhibition at 50 mM, 11% inhibition at 20 mM Thiobacillus thioparus
o-phenanthroline 37% inhibition at 10 mM Thiobacillus thioparus
oxaloacetate more than 50% inhibition at 20 mM Thiobacillus thioparus
p-hydroxymercuribenzoate complete inhibition at 1 mM, 5 min Thiobacillus thioparus
sodium arsenite 100% inhibition at 50 mM , 10% inhibition at 20 mM Thiobacillus thioparus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.003
-
NADPH
-
Thiobacillus thioparus
0.05
-
alpha-ketoglutarate
-
Thiobacillus thioparus
1.1
-
L-glutamine
-
Thiobacillus thioparus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
72000
-
x * 72000 + x * 94000 Thiobacillus thioparus
94000
-
x * 72000 + x * 94000 Thiobacillus thioparus
280000
-
gel filtration Thiobacillus thioparus

Organism

Organism UniProt Comment Textmining
Thiobacillus thioparus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
using ammonium sulfate fractionation, column chromatography on DEAE-cellulose and Sephadex G-200 Thiobacillus thioparus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
-
Thiobacillus thioparus
9.3
-
-
Thiobacillus thioparus

Storage Stability

Storage Stability Organism
storing at -20°C, total loss of activity after 10 days Thiobacillus thioparus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-glutamine + 2-oxoglutarate + NADPH + H+ in the reverse reaction ammonia can act instead of glutamine, but more slowly Thiobacillus thioparus L-glutamate + NADP+
-
?
L-glutamine + 2-oxoglutarate + NADPH + H+ NH3-dependent activity is increased approximately 5-fold in apoglutamate synthase lacking flavin and non-heme iron Thiobacillus thioparus L-glutamate + NADP+
-
?
additional information alpha-ketoglutarate can not be replaced with pyruvate or oxalacetate Thiobacillus thioparus ?
-
?
NH3 + 2-oxoglutarate + NADPH + H+ ammonia activity with 100 mM NH4Cl is about 6% of the glutamine activity with 5 mM L-glutamine Thiobacillus thioparus L-glutamate + NADP+
-
?

Subunits

Subunits Comment Organism
? x * 72000 + x * 94000 Thiobacillus thioparus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
-20
-
complete loss of activity after 10 days Thiobacillus thioparus
4
-
unprotected enzyme loses 90% of activity in 48 hours, protected enzyme, addition of 5 mM EDTA and 5 mM 2-mercaptoethanol or 1 mM DTT retains 80 to 90% of the activity after 48 hours Thiobacillus thioparus
100
-
30 s, complete loss of activity Thiobacillus thioparus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.3 7.8
-
Thiobacillus thioparus

Cofactor

Cofactor Comment Organism Structure
flavin flavoenzyme Thiobacillus thioparus
NADH no activity with NADH Thiobacillus thioparus
NADPH
-
Thiobacillus thioparus