BRENDA - Enzyme Database

Purification and properties of glutamate synthase from Thiobacillus thioparus

Adachi, K.; Suzuki, I.; J. Bacteriol. 129, 1173-1182 (1977)

Data extracted from this reference:

General Stability
EC Number
General Stability
Organism
1.4.1.13
2-mercaptoethanol, stabilizes
Thiobacillus thioparus
1.4.1.13
DTT, 1 mM, stabilizes at 4C
Thiobacillus thioparus
1.4.1.13
EDTA, stabilizes
Thiobacillus thioparus
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
1.4.1.13
2,2'-bipyridyl
100% inhibition at 3 mM
Thiobacillus thioparus
1.4.1.13
Atebrin
100% inhibition at 1 mM
Thiobacillus thioparus
1.4.1.13
ATP
33% inhibition at 20 mM
Thiobacillus thioparus
1.4.1.13
D-glutamate
-
Thiobacillus thioparus
1.4.1.13
iodoacetate
27% inhibition at 25 mM
Thiobacillus thioparus
1.4.1.13
isocitrate
32% inhibition at 20 mM
Thiobacillus thioparus
1.4.1.13
KCN
89% inhibition at 20 mM
Thiobacillus thioparus
1.4.1.13
L-glutamate
more than 40% inhibition at 50 mM
Thiobacillus thioparus
1.4.1.13
L-histidine
more than 40% inhibition at 50 mM
Thiobacillus thioparus
1.4.1.13
L-methionine
more than 40% inhibition at 50 mM
Thiobacillus thioparus
1.4.1.13
L-serine
more than 40% inhibition at 10 mM and 50 mM
Thiobacillus thioparus
1.4.1.13
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
1.4.1.13
N-ethylmaleimide
83% inhibition at 10 mM
Thiobacillus thioparus
1.4.1.13
NADP+
59% inhibition at 1 mM
Thiobacillus thioparus
1.4.1.13
NaN3
100% inhibition at 50 mM, 11% inhibition at 20 mM
Thiobacillus thioparus
1.4.1.13
o-phenanthroline
37% inhibition at 10 mM
Thiobacillus thioparus
1.4.1.13
oxaloacetate
more than 50% inhibition at 20 mM
Thiobacillus thioparus
1.4.1.13
p-hydroxymercuribenzoate
complete inhibition at 1 mM, 5 min
Thiobacillus thioparus
1.4.1.13
sodium arsenite
100% inhibition at 50 mM , 10% inhibition at 20 mM
Thiobacillus thioparus
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.4.1.13
0.003
-
NADPH
-
Thiobacillus thioparus
1.4.1.13
0.05
-
alpha-ketoglutarate
-
Thiobacillus thioparus
1.4.1.13
1.1
-
L-glutamine
-
Thiobacillus thioparus
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.4.1.13
72000
-
x * 72000 + x * 94000
Thiobacillus thioparus
1.4.1.13
94000
-
x * 72000 + x * 94000
Thiobacillus thioparus
1.4.1.13
280000
-
gel filtration
Thiobacillus thioparus
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.4.1.13
Thiobacillus thioparus
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.4.1.13
using ammonium sulfate fractionation, column chromatography on DEAE-cellulose and Sephadex G-200
Thiobacillus thioparus
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
1.4.1.13
additional information
-
-
Thiobacillus thioparus
1.4.1.13
9.3
-
-
Thiobacillus thioparus
Storage Stability
EC Number
Storage Stability
Organism
1.4.1.13
storing at -20C, total loss of activity after 10 days
Thiobacillus thioparus
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.4.1.13
L-glutamine + 2-oxoglutarate + NADPH + H+
in the reverse reaction ammonia can act instead of glutamine, but more slowly
391455
Thiobacillus thioparus
L-glutamate + NADP+
-
391455
Thiobacillus thioparus
?
1.4.1.13
L-glutamine + 2-oxoglutarate + NADPH + H+
NH3-dependent activity is increased approximately 5-fold in apoglutamate synthase lacking flavin and non-heme iron
391455
Thiobacillus thioparus
L-glutamate + NADP+
-
391455
Thiobacillus thioparus
?
1.4.1.13
additional information
alpha-ketoglutarate can not be replaced with pyruvate or oxalacetate
391455
Thiobacillus thioparus
?
-
-
-
-
1.4.1.13
NH3 + 2-oxoglutarate + NADPH + H+
ammonia activity with 100 mM NH4Cl is about 6% of the glutamine activity with 5 mM L-glutamine
391455
Thiobacillus thioparus
L-glutamate + NADP+
-
391455
Thiobacillus thioparus
?
Subunits
EC Number
Subunits
Commentary
Organism
1.4.1.13
?
x * 72000 + x * 94000
Thiobacillus thioparus
Temperature Stability [C]
EC Number
Temperature Stability Minimum [C]
Temperature Stability Maximum [C]
Commentary
Organism
1.4.1.13
-20
-
complete loss of activity after 10 days
Thiobacillus thioparus
1.4.1.13
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
1.4.1.13
100
-
30 s, complete loss of activity
Thiobacillus thioparus
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.4.1.13
7.3
7.8
-
Thiobacillus thioparus
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.4.1.13
flavin
flavoenzyme
Thiobacillus thioparus
1.4.1.13
NADH
no activity with NADH
Thiobacillus thioparus
1.4.1.13
NADPH
-
Thiobacillus thioparus
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.4.1.13
flavin
flavoenzyme
Thiobacillus thioparus
1.4.1.13
NADH
no activity with NADH
Thiobacillus thioparus
1.4.1.13
NADPH
-
Thiobacillus thioparus
General Stability (protein specific)
EC Number
General Stability
Organism
1.4.1.13
2-mercaptoethanol, stabilizes
Thiobacillus thioparus
1.4.1.13
DTT, 1 mM, stabilizes at 4C
Thiobacillus thioparus
1.4.1.13
EDTA, stabilizes
Thiobacillus thioparus
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
1.4.1.13
2,2'-bipyridyl
100% inhibition at 3 mM
Thiobacillus thioparus
1.4.1.13
Atebrin
100% inhibition at 1 mM
Thiobacillus thioparus
1.4.1.13
ATP
33% inhibition at 20 mM
Thiobacillus thioparus
1.4.1.13
D-glutamate
-
Thiobacillus thioparus
1.4.1.13
iodoacetate
27% inhibition at 25 mM
Thiobacillus thioparus
1.4.1.13
isocitrate
32% inhibition at 20 mM
Thiobacillus thioparus
1.4.1.13
KCN
89% inhibition at 20 mM
Thiobacillus thioparus
1.4.1.13
L-glutamate
more than 40% inhibition at 50 mM
Thiobacillus thioparus
1.4.1.13
L-histidine
more than 40% inhibition at 50 mM
Thiobacillus thioparus
1.4.1.13
L-methionine
more than 40% inhibition at 50 mM
Thiobacillus thioparus
1.4.1.13
L-serine
more than 40% inhibition at 10 mM and 50 mM
Thiobacillus thioparus
1.4.1.13
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
1.4.1.13
N-ethylmaleimide
83% inhibition at 10 mM
Thiobacillus thioparus
1.4.1.13
NADP+
59% inhibition at 1 mM
Thiobacillus thioparus
1.4.1.13
NaN3
100% inhibition at 50 mM, 11% inhibition at 20 mM
Thiobacillus thioparus
1.4.1.13
o-phenanthroline
37% inhibition at 10 mM
Thiobacillus thioparus
1.4.1.13
oxaloacetate
more than 50% inhibition at 20 mM
Thiobacillus thioparus
1.4.1.13
p-hydroxymercuribenzoate
complete inhibition at 1 mM, 5 min
Thiobacillus thioparus
1.4.1.13
sodium arsenite
100% inhibition at 50 mM , 10% inhibition at 20 mM
Thiobacillus thioparus
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.4.1.13
0.003
-
NADPH
-
Thiobacillus thioparus
1.4.1.13
0.05
-
alpha-ketoglutarate
-
Thiobacillus thioparus
1.4.1.13
1.1
-
L-glutamine
-
Thiobacillus thioparus
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.4.1.13
72000
-
x * 72000 + x * 94000
Thiobacillus thioparus
1.4.1.13
94000
-
x * 72000 + x * 94000
Thiobacillus thioparus
1.4.1.13
280000
-
gel filtration
Thiobacillus thioparus
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.4.1.13
using ammonium sulfate fractionation, column chromatography on DEAE-cellulose and Sephadex G-200
Thiobacillus thioparus
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
1.4.1.13
additional information
-
-
Thiobacillus thioparus
1.4.1.13
9.3
-
-
Thiobacillus thioparus
Storage Stability (protein specific)
EC Number
Storage Stability
Organism
1.4.1.13
storing at -20C, total loss of activity after 10 days
Thiobacillus thioparus
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.4.1.13
L-glutamine + 2-oxoglutarate + NADPH + H+
in the reverse reaction ammonia can act instead of glutamine, but more slowly
391455
Thiobacillus thioparus
L-glutamate + NADP+
-
391455
Thiobacillus thioparus
?
1.4.1.13
L-glutamine + 2-oxoglutarate + NADPH + H+
NH3-dependent activity is increased approximately 5-fold in apoglutamate synthase lacking flavin and non-heme iron
391455
Thiobacillus thioparus
L-glutamate + NADP+
-
391455
Thiobacillus thioparus
?
1.4.1.13
additional information
alpha-ketoglutarate can not be replaced with pyruvate or oxalacetate
391455
Thiobacillus thioparus
?
-
-
-
-
1.4.1.13
NH3 + 2-oxoglutarate + NADPH + H+
ammonia activity with 100 mM NH4Cl is about 6% of the glutamine activity with 5 mM L-glutamine
391455
Thiobacillus thioparus
L-glutamate + NADP+
-
391455
Thiobacillus thioparus
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.4.1.13
?
x * 72000 + x * 94000
Thiobacillus thioparus
Temperature Stability [C] (protein specific)
EC Number
Temperature Stability Minimum [C]
Temperature Stability Maximum [C]
Commentary
Organism
1.4.1.13
-20
-
complete loss of activity after 10 days
Thiobacillus thioparus
1.4.1.13
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
1.4.1.13
100
-
30 s, complete loss of activity
Thiobacillus thioparus
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.4.1.13
7.3
7.8
-
Thiobacillus thioparus