BRENDA - Enzyme Database

Gene cloning and characterization of the very large NAD-dependent L-glutamate dehydrogenase from the psychrophile Janthinobacterium lividum, isolated from cold soil

Kawakami, R.; Sakuraba, H.; Ohshima, T.; J. Bacteriol. 189, 5626-5633 (2007)

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
1.4.1.2
D-arginine
212% activity in the presence of 10 mM D-arginine
Janthinobacterium lividum
1.4.1.2
L-arginine
936% activity in the presence of 10 mM L-arginine
Janthinobacterium lividum
1.4.1.2
L-asparagine
118% activity in the presence of 10 mM L-asparagine
Janthinobacterium lividum
1.4.1.2
L-aspartate
1735% activity in the presence of 10 mM L-aspartate
Janthinobacterium lividum
1.4.1.2
L-histidine
155% activity in the presence of 10 mM L-histidine
Janthinobacterium lividum
1.4.1.2
L-lysine
161% activity in the presence of 10 mM L-lysine
Janthinobacterium lividum
1.4.1.2
L-methionine
206% activity in the presence of 10 mM L-methionine
Janthinobacterium lividum
1.4.1.2
L-tryptophan
235% activity in the presence of 10 mM L-tryptophan
Janthinobacterium lividum
1.4.1.2
Zn2+
224% activity in the presence of 1 mM Zn2+
Janthinobacterium lividum
Cloned(Commentary)
EC Number
Commentary
Organism
1.4.1.2
expressed in Escherichia coli Top10 cells
Janthinobacterium lividum
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
1.4.1.2
citrate
67% activity in the presence of 10 mM citrate
Janthinobacterium lividum
1.4.1.2
Co2+
82% activity in the presence of 1 mM Co2+
Janthinobacterium lividum
1.4.1.2
D-asparagine
93% activity in the presence of 10 mM D-asparagine
Janthinobacterium lividum
1.4.1.2
D-Aspartate
90% activity in the presence of 10 mM D-aspartate
Janthinobacterium lividum
1.4.1.2
D-glutamate
94% activity in the presence of 10 mM D-glutamate
Janthinobacterium lividum
1.4.1.2
D-glutamine
95% activity in the presence of 10 mM D-glutamine
Janthinobacterium lividum
1.4.1.2
fumarate
32% activity in the presence of 10 mM fumarate
Janthinobacterium lividum
1.4.1.2
Hg2+
no activity in the presence of 1 mM Hg2+
Janthinobacterium lividum
1.4.1.2
isocitrate
49% activity in the presence of 10 mM isocitrate
Janthinobacterium lividum
1.4.1.2
L-glutamate
substrate inhibition at L-glutamate concentrations above 20 mM
Janthinobacterium lividum
1.4.1.2
L-glutamine
89% activity in the presence of 10 mM L-glutamine
Janthinobacterium lividum
1.4.1.2
L-ornithine
83% activity in the presence of 10 mM L-ornithine
Janthinobacterium lividum
1.4.1.2
malate
65% activity in the presence of 10 mM malate
Janthinobacterium lividum
1.4.1.2
Mg2+
64% activity in the presence of 1 mM Mg2+
Janthinobacterium lividum
1.4.1.2
Mn2+
52% activity in the presence of 1 mM Mn2+
Janthinobacterium lividum
1.4.1.2
additional information
not inhibited by NAD+
Janthinobacterium lividum
1.4.1.2
Ni2+
77% activity in the presence of 1 mM Ni2+
Janthinobacterium lividum
1.4.1.2
succinate
36% activity in the presence of 10 mM succinate
Janthinobacterium lividum
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.4.1.2
2.1
-
NAD+
in the presence of 20 mM L-glutamate, in 100 mM glycine/NaOH (pH 9.5), at 25C
Janthinobacterium lividum
1.4.1.2
7.1
-
L-glutamate
in the presence of 5 mM NAD+, in 100 mM glycine/NaOH (pH 9.5), at 25C
Janthinobacterium lividum
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.4.1.2
169360
-
6 * 169360, deduced from amino acid sequence
Janthinobacterium lividum
1.4.1.2
170000
-
6 * 170000, SDS-PAGE
Janthinobacterium lividum
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.4.1.2
Janthinobacterium lividum
A5LH94
fragment
-
1.4.1.2
Janthinobacterium lividum UTB1302
A5LH94
fragment
-
Purification (Commentary)
EC Number
Commentary
Organism
1.4.1.2
DEAE-Toyopearl column chromatography, butyl-Toyopearl column chromatography, and Superdex 200 gel filtration
Janthinobacterium lividum
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
1.4.1.2
0.62
-
recombinant enzyme from crude cell extract, at 25C
Janthinobacterium lividum
1.4.1.2
3.1
-
purified recombinant enzyme, at 25C
Janthinobacterium lividum
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.4.1.2
L-glutamate + H2O + NAD+
-
692832
Janthinobacterium lividum
2-oxoglutarate + NH3 + NADH
-
-
-
?
1.4.1.2
L-glutamate + H2O + NAD+
-
692832
Janthinobacterium lividum UTB1302
2-oxoglutarate + NH3 + NADH
-
-
-
?
1.4.1.2
additional information
inert reaction with L-glutamine, L-aspartate, L-alanine, L-leucine, L-valine, L-lysine, L-2-aminobutyrate, L-methionine, L-ornithine, L-phenylalanine, L-arginine, L-tryptophan, L-methionine, L-histidine, D-glutamate, and D-aspartate, the enzyme is not active with NADP+
692832
Janthinobacterium lividum
?
-
-
-
-
1.4.1.2
additional information
inert reaction with L-glutamine, L-aspartate, L-alanine, L-leucine, L-valine, L-lysine, L-2-aminobutyrate, L-methionine, L-ornithine, L-phenylalanine, L-arginine, L-tryptophan, L-methionine, L-histidine, D-glutamate, and D-aspartate, the enzyme is not active with NADP+
692832
Janthinobacterium lividum UTB1302
?
-
-
-
-
Subunits
EC Number
Subunits
Commentary
Organism
1.4.1.2
homohexamer
6 * 169360, deduced from amino acid sequence; 6 * 170000, SDS-PAGE
Janthinobacterium lividum
Temperature Optimum [C]
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
1.4.1.2
40
-
in glycine-NaOH buffer at pH 9.0
Janthinobacterium lividum
Temperature Stability [C]
EC Number
Temperature Stability Minimum [C]
Temperature Stability Maximum [C]
Commentary
Organism
1.4.1.2
40
50
the enzyme is stable for 10 min at temperatures up to 40C but is completely inactivated by incubation for 10 min at 50C
Janthinobacterium lividum
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.4.1.2
9
-
in glycine-NaOH buffer at 40C
Janthinobacterium lividum
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.4.1.2
NAD+
dependent
Janthinobacterium lividum
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
1.4.1.2
D-arginine
212% activity in the presence of 10 mM D-arginine
Janthinobacterium lividum
1.4.1.2
L-arginine
936% activity in the presence of 10 mM L-arginine
Janthinobacterium lividum
1.4.1.2
L-asparagine
118% activity in the presence of 10 mM L-asparagine
Janthinobacterium lividum
1.4.1.2
L-aspartate
1735% activity in the presence of 10 mM L-aspartate
Janthinobacterium lividum
1.4.1.2
L-histidine
155% activity in the presence of 10 mM L-histidine
Janthinobacterium lividum
1.4.1.2
L-lysine
161% activity in the presence of 10 mM L-lysine
Janthinobacterium lividum
1.4.1.2
L-methionine
206% activity in the presence of 10 mM L-methionine
Janthinobacterium lividum
1.4.1.2
L-tryptophan
235% activity in the presence of 10 mM L-tryptophan
Janthinobacterium lividum
1.4.1.2
Zn2+
224% activity in the presence of 1 mM Zn2+
Janthinobacterium lividum
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.4.1.2
expressed in Escherichia coli Top10 cells
Janthinobacterium lividum
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.4.1.2
NAD+
dependent
Janthinobacterium lividum
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
1.4.1.2
citrate
67% activity in the presence of 10 mM citrate
Janthinobacterium lividum
1.4.1.2
Co2+
82% activity in the presence of 1 mM Co2+
Janthinobacterium lividum
1.4.1.2
D-asparagine
93% activity in the presence of 10 mM D-asparagine
Janthinobacterium lividum
1.4.1.2
D-Aspartate
90% activity in the presence of 10 mM D-aspartate
Janthinobacterium lividum
1.4.1.2
D-glutamate
94% activity in the presence of 10 mM D-glutamate
Janthinobacterium lividum
1.4.1.2
D-glutamine
95% activity in the presence of 10 mM D-glutamine
Janthinobacterium lividum
1.4.1.2
fumarate
32% activity in the presence of 10 mM fumarate
Janthinobacterium lividum
1.4.1.2
Hg2+
no activity in the presence of 1 mM Hg2+
Janthinobacterium lividum
1.4.1.2
isocitrate
49% activity in the presence of 10 mM isocitrate
Janthinobacterium lividum
1.4.1.2
L-glutamate
substrate inhibition at L-glutamate concentrations above 20 mM
Janthinobacterium lividum
1.4.1.2
L-glutamine
89% activity in the presence of 10 mM L-glutamine
Janthinobacterium lividum
1.4.1.2
L-ornithine
83% activity in the presence of 10 mM L-ornithine
Janthinobacterium lividum
1.4.1.2
malate
65% activity in the presence of 10 mM malate
Janthinobacterium lividum
1.4.1.2
Mg2+
64% activity in the presence of 1 mM Mg2+
Janthinobacterium lividum
1.4.1.2
Mn2+
52% activity in the presence of 1 mM Mn2+
Janthinobacterium lividum
1.4.1.2
additional information
not inhibited by NAD+
Janthinobacterium lividum
1.4.1.2
Ni2+
77% activity in the presence of 1 mM Ni2+
Janthinobacterium lividum
1.4.1.2
succinate
36% activity in the presence of 10 mM succinate
Janthinobacterium lividum
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.4.1.2
2.1
-
NAD+
in the presence of 20 mM L-glutamate, in 100 mM glycine/NaOH (pH 9.5), at 25C
Janthinobacterium lividum
1.4.1.2
7.1
-
L-glutamate
in the presence of 5 mM NAD+, in 100 mM glycine/NaOH (pH 9.5), at 25C
Janthinobacterium lividum
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.4.1.2
169360
-
6 * 169360, deduced from amino acid sequence
Janthinobacterium lividum
1.4.1.2
170000
-
6 * 170000, SDS-PAGE
Janthinobacterium lividum
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.4.1.2
DEAE-Toyopearl column chromatography, butyl-Toyopearl column chromatography, and Superdex 200 gel filtration
Janthinobacterium lividum
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
1.4.1.2
0.62
-
recombinant enzyme from crude cell extract, at 25C
Janthinobacterium lividum
1.4.1.2
3.1
-
purified recombinant enzyme, at 25C
Janthinobacterium lividum
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.4.1.2
L-glutamate + H2O + NAD+
-
692832
Janthinobacterium lividum
2-oxoglutarate + NH3 + NADH
-
-
-
?
1.4.1.2
L-glutamate + H2O + NAD+
-
692832
Janthinobacterium lividum UTB1302
2-oxoglutarate + NH3 + NADH
-
-
-
?
1.4.1.2
additional information
inert reaction with L-glutamine, L-aspartate, L-alanine, L-leucine, L-valine, L-lysine, L-2-aminobutyrate, L-methionine, L-ornithine, L-phenylalanine, L-arginine, L-tryptophan, L-methionine, L-histidine, D-glutamate, and D-aspartate, the enzyme is not active with NADP+
692832
Janthinobacterium lividum
?
-
-
-
-
1.4.1.2
additional information
inert reaction with L-glutamine, L-aspartate, L-alanine, L-leucine, L-valine, L-lysine, L-2-aminobutyrate, L-methionine, L-ornithine, L-phenylalanine, L-arginine, L-tryptophan, L-methionine, L-histidine, D-glutamate, and D-aspartate, the enzyme is not active with NADP+
692832
Janthinobacterium lividum UTB1302
?
-
-
-
-
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.4.1.2
homohexamer
6 * 169360, deduced from amino acid sequence; 6 * 170000, SDS-PAGE
Janthinobacterium lividum
Temperature Optimum [C] (protein specific)
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
1.4.1.2
40
-
in glycine-NaOH buffer at pH 9.0
Janthinobacterium lividum
Temperature Stability [C] (protein specific)
EC Number
Temperature Stability Minimum [C]
Temperature Stability Maximum [C]
Commentary
Organism
1.4.1.2
40
50
the enzyme is stable for 10 min at temperatures up to 40C but is completely inactivated by incubation for 10 min at 50C
Janthinobacterium lividum
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.4.1.2
9
-
in glycine-NaOH buffer at 40C
Janthinobacterium lividum