BRENDA - Enzyme Database

Purification and properties of glutamate synthase from Nocardia mediterranei

Mei, B.; Jiao, R.; J. Bacteriol. 170, 1940-1944 (1988)

Data extracted from this reference:

General Stability
EC Number
General Stability
Organism
1.4.1.13
2 mM alpha-ketoglutarate, 10 mM beta-mercaptoethanol and 10% glycerol when present together markedly enhance stability
Amycolatopsis mediterranei
1.4.1.13
unprotected enzyme is unstable, losing 65% of its activity in 24 hours at 4°C
Amycolatopsis mediterranei
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
1.4.1.13
D-glutamate
-
Amycolatopsis mediterranei
1.4.1.13
glycine
less than 50% inhibition at 5 mM
Amycolatopsis mediterranei
1.4.1.13
isocitrate
35% inhibition at 5 mM
Amycolatopsis mediterranei
1.4.1.13
L-alanine
less than 50% inhibition at 5 mM
Amycolatopsis mediterranei
1.4.1.13
L-asparagine
less than 50% inhibition at 5 mM
Amycolatopsis mediterranei
1.4.1.13
L-glutamate
potent inhibitor
Amycolatopsis mediterranei
1.4.1.13
L-histidine
less than 50% inhibition at 5 mM
Amycolatopsis mediterranei
1.4.1.13
L-leucine
less than 50% inhibition at 5 mM
Amycolatopsis mediterranei
1.4.1.13
L-malate
more than 50% inhibition at 5 mM
Amycolatopsis mediterranei
1.4.1.13
L-methionine
shows no effect
Amycolatopsis mediterranei
1.4.1.13
L-serine
less than 50% inhibition at 5 mM
Amycolatopsis mediterranei
1.4.1.13
L-tryptophan
strong inhibitor
Amycolatopsis mediterranei
1.4.1.13
NADP+
potent inhibitor
Amycolatopsis mediterranei
1.4.1.13
succinate
25% inhibition at 5 mM
Amycolatopsis mediterranei
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.4.1.13
0.053
-
alpha-ketoglutarate
-
Amycolatopsis mediterranei
1.4.1.13
0.077
-
L-glutamine
-
Amycolatopsis mediterranei
1.4.1.13
0.11
-
NADPH
-
Amycolatopsis mediterranei
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
1.4.1.13
Iron
contains 7.5 mol of nonheme iron; iron-sulfur protein
Amycolatopsis mediterranei
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.4.1.13
55000
-
1 * 145000 + 1 * 55000, SDS-PAGE
Amycolatopsis mediterranei
1.4.1.13
145000
-
1 * 145000 + 1 * 55000, SDS-PAGE
Amycolatopsis mediterranei
1.4.1.13
195000
-
sucrose density gradient centrifugation
Amycolatopsis mediterranei
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.4.1.13
Amycolatopsis mediterranei
-
-
-
Oxidation Stability
EC Number
Oxidation Stability
Organism
1.4.1.13
presence of reducing agents is essential for stabilization
Amycolatopsis mediterranei
Purification (Commentary)
EC Number
Commentary
Organism
1.4.1.13
using protamine sulfate precipitation, ammonium sulfate precipitation, column chromatography on DE52, DEAE-Sephadex, hydroxyapatite, phenyl-Sepharose CL-4B and Bio-Gel filtration
Amycolatopsis mediterranei
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.4.1.13
1.48
-
-
Amycolatopsis mediterranei
Storage Stability
EC Number
Storage Stability
Organism
1.4.1.13
4°C, unprotected enzyme loses 65% of activity after 24 h
Amycolatopsis mediterranei
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
391440
Amycolatopsis mediterranei
L-glutamate + NADP+
-
391440
Amycolatopsis mediterranei
?
1.4.1.13
NH3 + 2-oxoglutarate + NADPH + H+
14% relative activity to L-glutamine
391440
Amycolatopsis mediterranei
L-glutamate + NADP+
-
391440
Amycolatopsis mediterranei
?
Subunits
EC Number
Subunits
Commentary
Organism
1.4.1.13
dimer
1 * 145000 + 1 * 55000, SDS-PAGE
Amycolatopsis mediterranei
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.4.1.13
7.5
8.5
broad optimum
Amycolatopsis mediterranei
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.4.1.13
flavin
contains FAD and FMN; ratio FAD: FMN is 1.1
Amycolatopsis mediterranei
1.4.1.13
additional information
the enzyme contains 7.2 mol of acid-labile sulfur per 200,000 g of protein
Amycolatopsis mediterranei
1.4.1.13
NADH
no activity with NADH
Amycolatopsis mediterranei
1.4.1.13
NADPH
-
Amycolatopsis mediterranei
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.4.1.13
flavin
contains FAD and FMN; ratio FAD: FMN is 1.1
Amycolatopsis mediterranei
1.4.1.13
additional information
the enzyme contains 7.2 mol of acid-labile sulfur per 200,000 g of protein
Amycolatopsis mediterranei
1.4.1.13
NADH
no activity with NADH
Amycolatopsis mediterranei
1.4.1.13
NADPH
-
Amycolatopsis mediterranei
General Stability (protein specific)
EC Number
General Stability
Organism
1.4.1.13
2 mM alpha-ketoglutarate, 10 mM beta-mercaptoethanol and 10% glycerol when present together markedly enhance stability
Amycolatopsis mediterranei
1.4.1.13
unprotected enzyme is unstable, losing 65% of its activity in 24 hours at 4°C
Amycolatopsis mediterranei
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
1.4.1.13
D-glutamate
-
Amycolatopsis mediterranei
1.4.1.13
glycine
less than 50% inhibition at 5 mM
Amycolatopsis mediterranei
1.4.1.13
isocitrate
35% inhibition at 5 mM
Amycolatopsis mediterranei
1.4.1.13
L-alanine
less than 50% inhibition at 5 mM
Amycolatopsis mediterranei
1.4.1.13
L-asparagine
less than 50% inhibition at 5 mM
Amycolatopsis mediterranei
1.4.1.13
L-glutamate
potent inhibitor
Amycolatopsis mediterranei
1.4.1.13
L-histidine
less than 50% inhibition at 5 mM
Amycolatopsis mediterranei
1.4.1.13
L-leucine
less than 50% inhibition at 5 mM
Amycolatopsis mediterranei
1.4.1.13
L-malate
more than 50% inhibition at 5 mM
Amycolatopsis mediterranei
1.4.1.13
L-methionine
shows no effect
Amycolatopsis mediterranei
1.4.1.13
L-serine
less than 50% inhibition at 5 mM
Amycolatopsis mediterranei
1.4.1.13
L-tryptophan
strong inhibitor
Amycolatopsis mediterranei
1.4.1.13
NADP+
potent inhibitor
Amycolatopsis mediterranei
1.4.1.13
succinate
25% inhibition at 5 mM
Amycolatopsis mediterranei
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.4.1.13
0.053
-
alpha-ketoglutarate
-
Amycolatopsis mediterranei
1.4.1.13
0.077
-
L-glutamine
-
Amycolatopsis mediterranei
1.4.1.13
0.11
-
NADPH
-
Amycolatopsis mediterranei
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
1.4.1.13
Iron
contains 7.5 mol of nonheme iron; iron-sulfur protein
Amycolatopsis mediterranei
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.4.1.13
55000
-
1 * 145000 + 1 * 55000, SDS-PAGE
Amycolatopsis mediterranei
1.4.1.13
145000
-
1 * 145000 + 1 * 55000, SDS-PAGE
Amycolatopsis mediterranei
1.4.1.13
195000
-
sucrose density gradient centrifugation
Amycolatopsis mediterranei
Oxidation Stability (protein specific)
EC Number
Oxidation Stability
Organism
1.4.1.13
presence of reducing agents is essential for stabilization
Amycolatopsis mediterranei
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.4.1.13
using protamine sulfate precipitation, ammonium sulfate precipitation, column chromatography on DE52, DEAE-Sephadex, hydroxyapatite, phenyl-Sepharose CL-4B and Bio-Gel filtration
Amycolatopsis mediterranei
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.4.1.13
1.48
-
-
Amycolatopsis mediterranei
Storage Stability (protein specific)
EC Number
Storage Stability
Organism
1.4.1.13
4°C, unprotected enzyme loses 65% of activity after 24 h
Amycolatopsis mediterranei
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
391440
Amycolatopsis mediterranei
L-glutamate + NADP+
-
391440
Amycolatopsis mediterranei
?
1.4.1.13
NH3 + 2-oxoglutarate + NADPH + H+
14% relative activity to L-glutamine
391440
Amycolatopsis mediterranei
L-glutamate + NADP+
-
391440
Amycolatopsis mediterranei
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.4.1.13
dimer
1 * 145000 + 1 * 55000, SDS-PAGE
Amycolatopsis mediterranei
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.4.1.13
7.5
8.5
broad optimum
Amycolatopsis mediterranei