BRENDA - Enzyme Database

Biochemical characterization of NADP+-dependent isocitrate dehydrogenase from Microcystis aeruginosa PCC7806

Jin, M.M.; Wang, P.; Li, X.; Zhao, X.Y.; Xu, L.; Song, P.; Zhu, G.P.; Mol. Biol. Rep. 40, 2995-3002 (2013)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
1.1.1.42
gene icd, sequence comparisons, expression of His6-tagged in Escherichia coli strain Rosetta (DE3); gene icd, sequence comparisons, recombinant expression of His6-tagged enzyme in Escherichia coli strain Rosetta (DE3)
Microcystis aeruginosa
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
1.1.1.42
Ca2+
-
Microcystis aeruginosa
1.1.1.42
Co2+
-
Microcystis aeruginosa
1.1.1.42
Cu2+
complete inhibition
Microcystis aeruginosa
1.1.1.42
K+
slight inhibition
Microcystis aeruginosa
1.1.1.42
Li+
slight inhibition
Microcystis aeruginosa
1.1.1.42
Na+
slight inhibition
Microcystis aeruginosa
1.1.1.42
Ni2+
-
Microcystis aeruginosa
1.1.1.42
Rb+
slight inhibition
Microcystis aeruginosa
1.1.1.42
Zn2+
strong inhibition
Microcystis aeruginosa
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.42
additional information
-
additional information
kinetic analysis
Microcystis aeruginosa
1.1.1.42
0.0322
-
NADP+
with Mg2+, pH 7.5, 37C
Microcystis aeruginosa
1.1.1.42
0.0324
-
NADP+
with Mg2+, pH 7.5, 50C
Microcystis aeruginosa
1.1.1.42
0.0326
-
DL-isocitrate
with Mg2+, pH 7.5, 37C; with Mg2+, pH 7.5, 50C
Microcystis aeruginosa
1.1.1.42
0.0717
-
NADP+
with Mn2+, pH 7.5, 50C
Microcystis aeruginosa
1.1.1.42
0.0797
-
NADP+
with Mn2+, pH 7.5, 37C
Microcystis aeruginosa
1.1.1.42
0.1243
-
DL-isocitrate
with Mn2+, pH 7.5, 37C; with Mn2+, pH 7.5, 50C
Microcystis aeruginosa
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
1.1.1.42
Ca2+
activates poorly, 2.48% activity compared to Mn2+
Microcystis aeruginosa
1.1.1.42
Co2+
activates, 10.22% activity compared to Mn2+
Microcystis aeruginosa
1.1.1.42
K+
activates slightly, 10.18% activity compared to Mn2+
Microcystis aeruginosa
1.1.1.42
Li+
activates slightly, 12.63% activity compared to Mn2+
Microcystis aeruginosa
1.1.1.42
Mg2+
activates, 72.45% activity compared to Mn2+; can partially substitute for Mn2+
Microcystis aeruginosa
1.1.1.42
Mn2+
activates, required, best divalent cation; required
Microcystis aeruginosa
1.1.1.42
additional information
the enzyme is completely divalent cation dependent. As compared with Mn2+, MaIDH shows about 2.5times and 4times higher affinities (1/Km) to NADP+ and DL-isocitrate with Mg2+, Cu2+ does not activate the enzyme at all
Microcystis aeruginosa
1.1.1.42
Na+
activates slightly, 10.70% activity compared to Mn2+
Microcystis aeruginosa
1.1.1.42
Ni2+
activates poorly, 1.40% activity compared to Mn2+
Microcystis aeruginosa
1.1.1.42
Rb+
activates slightly, 20.67% activity compared to Mn2+
Microcystis aeruginosa
1.1.1.42
Zn2+
activates poorly, 4.23% activity compared to Mn2+
Microcystis aeruginosa
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.1.1.42
52600
-
; 2 * 52600
Microcystis aeruginosa
1.1.1.42
105000
-
gel filtration; gel filtration
Microcystis aeruginosa
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.1.1.42
DL-isocitrate + NADP+
Microcystis aeruginosa
-
2-oxoglutarate + CO2 + NADPH + H+
-
-
?
1.1.1.42
DL-isocitrate + NADP+
Microcystis aeruginosa
-
2-oxoglutarate + CO2 + NADPH + H+
-
-
r
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.1.1.42
Microcystis aeruginosa
A8YGS2
-
-
1.1.1.42
Microcystis aeruginosa
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.1.1.42
recombinant His6-tagged enzyme from Escherichia coli strain Rosetta (DE3) by metal affinity chromatography
Microcystis aeruginosa
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.42
DL-isocitrate + NADP+
-
723152
Microcystis aeruginosa
2-oxoglutarate + CO2 + NADPH + H+
-
-
-
?
1.1.1.42
DL-isocitrate + NADP+
-
723152
Microcystis aeruginosa
2-oxoglutarate + CO2 + NADPH + H+
-
-
-
r
Subunits
EC Number
Subunits
Commentary
Organism
1.1.1.42
homodimer
2 * 52600; 2 * 52600, SDS-PAGE
Microcystis aeruginosa
Temperature Optimum [C]
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
1.1.1.42
45
-
with Mg2+; with Mn2+
Microcystis aeruginosa
1.1.1.42
50
-
with Mg2+; with Mn2+
Microcystis aeruginosa
Temperature Range [C]
EC Number
Temperature Minimum [C]
Temperature Maximum [C]
Commentary
Organism
1.1.1.42
25
60
activity range
Microcystis aeruginosa
Temperature Stability [C]
EC Number
Temperature Stability Minimum [C]
Temperature Stability Maximum [C]
Commentary
Organism
1.1.1.42
30
-
purified recombinant enzyme, 20 min, stable
Microcystis aeruginosa
1.1.1.42
40
-
purified recombinant enzyme, 20 min, loss of 50% of enzyme activity, with Mg2+
Microcystis aeruginosa
1.1.1.42
45
-
purified enzyme, 20 min, 50% loss of enzyme activity; purified recombinant enzyme, 20 min, loss of 50% of enzyme activity, with Mn2+
Microcystis aeruginosa
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.1.1.42
10.29
-
DL-isocitrate
with Mg2+, pH 7.5, 50C
Microcystis aeruginosa
1.1.1.42
10.92
-
DL-isocitrate
with Mg2+, pH 7.5, 37C
Microcystis aeruginosa
1.1.1.42
25.36
-
NADP+
with Mg2+, pH 7.5, 37C; with Mg2+, pH 7.5, 50C
Microcystis aeruginosa
1.1.1.42
43.21
-
DL-isocitrate
with Mn2+, pH 7.5, 37C; with Mn2+, pH 7.5, 50C
Microcystis aeruginosa
1.1.1.42
48.88
-
NADP+
with Mn2+, pH 7.5, 37C; with Mn2+, pH 7.5, 50C
Microcystis aeruginosa
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.42
7.5
-
the recombinant MaIDH has a broader pH optimum from pH 6.5 to pH 8.5; with Mn2+ or Mg2+
Microcystis aeruginosa
pH Range
EC Number
pH Minimum
pH Maximum
Commentary
Organism
1.1.1.42
5.5
9.5
activity range
Microcystis aeruginosa
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.42
NADP+
purified recombinant MaIDH is completely specific with NADP+, and no NAD+-dependent activity is detectable; the enzyme is completely NADP+-dependent and no NAD+-linked activity is detectable
Microcystis aeruginosa
1.1.1.42
NADPH
-
Microcystis aeruginosa
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.42
gene icd, sequence comparisons, expression of His6-tagged in Escherichia coli strain Rosetta (DE3)
Microcystis aeruginosa
1.1.1.42
gene icd, sequence comparisons, recombinant expression of His6-tagged enzyme in Escherichia coli strain Rosetta (DE3)
Microcystis aeruginosa
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.42
NADP+
purified recombinant MaIDH is completely specific with NADP+, and no NAD+-dependent activity is detectable
Microcystis aeruginosa
1.1.1.42
NADP+
the enzyme is completely NADP+-dependent and no NAD+-linked activity is detectable
Microcystis aeruginosa
1.1.1.42
NADPH
-
Microcystis aeruginosa
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
1.1.1.42
Ca2+
-
Microcystis aeruginosa
1.1.1.42
Co2+
-
Microcystis aeruginosa
1.1.1.42
Cu2+
complete inhibition
Microcystis aeruginosa
1.1.1.42
K+
slight inhibition
Microcystis aeruginosa
1.1.1.42
Li+
slight inhibition
Microcystis aeruginosa
1.1.1.42
Na+
slight inhibition
Microcystis aeruginosa
1.1.1.42
Ni2+
-
Microcystis aeruginosa
1.1.1.42
Rb+
slight inhibition
Microcystis aeruginosa
1.1.1.42
Zn2+
strong inhibition
Microcystis aeruginosa
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.42
additional information
-
additional information
kinetic analysis
Microcystis aeruginosa
1.1.1.42
0.0322
-
NADP+
with Mg2+, pH 7.5, 37C
Microcystis aeruginosa
1.1.1.42
0.0324
-
NADP+
with Mg2+, pH 7.5, 50C
Microcystis aeruginosa
1.1.1.42
0.0326
-
DL-isocitrate
with Mg2+, pH 7.5, 50C
Microcystis aeruginosa
1.1.1.42
0.0326
-
DL-isocitrate
with Mg2+, pH 7.5, 37C
Microcystis aeruginosa
1.1.1.42
0.0717
-
NADP+
with Mn2+, pH 7.5, 50C
Microcystis aeruginosa
1.1.1.42
0.0797
-
NADP+
with Mn2+, pH 7.5, 37C
Microcystis aeruginosa
1.1.1.42
0.1243
-
DL-isocitrate
with Mn2+, pH 7.5, 50C
Microcystis aeruginosa
1.1.1.42
0.1243
-
DL-isocitrate
with Mn2+, pH 7.5, 37C
Microcystis aeruginosa
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
1.1.1.42
Ca2+
activates poorly, 2.48% activity compared to Mn2+
Microcystis aeruginosa
1.1.1.42
Co2+
activates, 10.22% activity compared to Mn2+
Microcystis aeruginosa
1.1.1.42
K+
activates slightly, 10.18% activity compared to Mn2+
Microcystis aeruginosa
1.1.1.42
Li+
activates slightly, 12.63% activity compared to Mn2+
Microcystis aeruginosa
1.1.1.42
Mg2+
can partially substitute for Mn2+
Microcystis aeruginosa
1.1.1.42
Mg2+
activates, 72.45% activity compared to Mn2+
Microcystis aeruginosa
1.1.1.42
Mn2+
required
Microcystis aeruginosa
1.1.1.42
Mn2+
activates, required, best divalent cation
Microcystis aeruginosa
1.1.1.42
additional information
the enzyme is completely divalent cation dependent. As compared with Mn2+, MaIDH shows about 2.5times and 4times higher affinities (1/Km) to NADP+ and DL-isocitrate with Mg2+, Cu2+ does not activate the enzyme at all
Microcystis aeruginosa
1.1.1.42
Na+
activates slightly, 10.70% activity compared to Mn2+
Microcystis aeruginosa
1.1.1.42
Ni2+
activates poorly, 1.40% activity compared to Mn2+
Microcystis aeruginosa
1.1.1.42
Rb+
activates slightly, 20.67% activity compared to Mn2+
Microcystis aeruginosa
1.1.1.42
Zn2+
activates poorly, 4.23% activity compared to Mn2+
Microcystis aeruginosa
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.1.1.42
52600
-
2 * 52600
Microcystis aeruginosa
1.1.1.42
52600
-
-
Microcystis aeruginosa
1.1.1.42
105000
-
gel filtration
Microcystis aeruginosa
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.1.1.42
DL-isocitrate + NADP+
Microcystis aeruginosa
-
2-oxoglutarate + CO2 + NADPH + H+
-
-
?
1.1.1.42
DL-isocitrate + NADP+
Microcystis aeruginosa
-
2-oxoglutarate + CO2 + NADPH + H+
-
-
r
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.42
recombinant His6-tagged enzyme from Escherichia coli strain Rosetta (DE3) by metal affinity chromatography
Microcystis aeruginosa
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.42
DL-isocitrate + NADP+
-
723152
Microcystis aeruginosa
2-oxoglutarate + CO2 + NADPH + H+
-
-
-
?
1.1.1.42
DL-isocitrate + NADP+
-
723152
Microcystis aeruginosa
2-oxoglutarate + CO2 + NADPH + H+
-
-
-
r
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.1.1.42
homodimer
2 * 52600
Microcystis aeruginosa
1.1.1.42
homodimer
2 * 52600, SDS-PAGE
Microcystis aeruginosa
Temperature Optimum [C] (protein specific)
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
1.1.1.42
45
-
with Mg2+
Microcystis aeruginosa
1.1.1.42
45
-
with Mn2+
Microcystis aeruginosa
1.1.1.42
50
-
with Mn2+
Microcystis aeruginosa
1.1.1.42
50
-
with Mg2+
Microcystis aeruginosa
Temperature Range [C] (protein specific)
EC Number
Temperature Minimum [C]
Temperature Maximum [C]
Commentary
Organism
1.1.1.42
25
60
activity range
Microcystis aeruginosa
Temperature Stability [C] (protein specific)
EC Number
Temperature Stability Minimum [C]
Temperature Stability Maximum [C]
Commentary
Organism
1.1.1.42
30
-
purified recombinant enzyme, 20 min, stable
Microcystis aeruginosa
1.1.1.42
40
-
purified recombinant enzyme, 20 min, loss of 50% of enzyme activity, with Mg2+
Microcystis aeruginosa
1.1.1.42
45
-
purified recombinant enzyme, 20 min, loss of 50% of enzyme activity, with Mn2+
Microcystis aeruginosa
1.1.1.42
45
-
purified enzyme, 20 min, 50% loss of enzyme activity
Microcystis aeruginosa
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.1.1.42
10.29
-
DL-isocitrate
with Mg2+, pH 7.5, 50C
Microcystis aeruginosa
1.1.1.42
10.92
-
DL-isocitrate
with Mg2+, pH 7.5, 37C
Microcystis aeruginosa
1.1.1.42
25.36
-
NADP+
with Mg2+, pH 7.5, 50C
Microcystis aeruginosa
1.1.1.42
25.36
-
NADP+
with Mg2+, pH 7.5, 37C
Microcystis aeruginosa
1.1.1.42
43.21
-
DL-isocitrate
with Mn2+, pH 7.5, 50C
Microcystis aeruginosa
1.1.1.42
43.21
-
DL-isocitrate
with Mn2+, pH 7.5, 37C
Microcystis aeruginosa
1.1.1.42
48.88
-
NADP+
with Mn2+, pH 7.5, 50C
Microcystis aeruginosa
1.1.1.42
48.88
-
NADP+
with Mn2+, pH 7.5, 37C
Microcystis aeruginosa
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.42
7.5
-
with Mn2+ or Mg2+
Microcystis aeruginosa
1.1.1.42
7.5
-
the recombinant MaIDH has a broader pH optimum from pH 6.5 to pH 8.5
Microcystis aeruginosa
pH Range (protein specific)
EC Number
pH Minimum
pH Maximum
Commentary
Organism
1.1.1.42
5.5
9.5
activity range
Microcystis aeruginosa
General Information
EC Number
General Information
Commentary
Organism
1.1.1.42
physiological function
isocitrate dehydrogenase of Microcystis aeruginosa plays important roles in energy and biosynthesis metabolisms and its catalytic product 2-oxoglutarate provides the carbon skeleton for ammonium assimilation and also constitutes a signaling molecule of nitrogen starvation in cyanobacteria
Microcystis aeruginosa
General Information (protein specific)
EC Number
General Information
Commentary
Organism
1.1.1.42
physiological function
isocitrate dehydrogenase of Microcystis aeruginosa plays important roles in energy and biosynthesis metabolisms and its catalytic product 2-oxoglutarate provides the carbon skeleton for ammonium assimilation and also constitutes a signaling molecule of nitrogen starvation in cyanobacteria
Microcystis aeruginosa
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.1.1.42
0.00335
-
DL-isocitrate
with Mg2+, pH 7.5, 50C
Microcystis aeruginosa
1.1.1.42
0.00348
-
DL-isocitrate
with Mn2+, pH 7.5, 50C
Microcystis aeruginosa
1.1.1.42
0.00613
-
NADP+
with Mn2+, pH 7.5, 50C
Microcystis aeruginosa
1.1.1.42
0.00787
-
NADP+
with Mg2+, pH 7.5, 50C
Microcystis aeruginosa
1.1.1.42
335
-
DL-isocitrate
with Mg2+, pH 7.5, 37C
Microcystis aeruginosa
1.1.1.42
348
-
DL-isocitrate
with Mn2+, pH 7.5, 37C
Microcystis aeruginosa
1.1.1.42
613
-
NADP+
with Mn2+, pH 7.5, 37C
Microcystis aeruginosa
1.1.1.42
787
-
NADP+
with Mg2+, pH 7.5, 37C
Microcystis aeruginosa
KCat/KM [mM/s] (protein specific)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.1.1.42
0.00335
-
DL-isocitrate
with Mg2+, pH 7.5, 50C
Microcystis aeruginosa
1.1.1.42
0.00348
-
DL-isocitrate
with Mn2+, pH 7.5, 50C
Microcystis aeruginosa
1.1.1.42
0.00613
-
NADP+
with Mn2+, pH 7.5, 50C
Microcystis aeruginosa
1.1.1.42
0.00787
-
NADP+
with Mg2+, pH 7.5, 50C
Microcystis aeruginosa
1.1.1.42
335
-
DL-isocitrate
with Mg2+, pH 7.5, 37C
Microcystis aeruginosa
1.1.1.42
348
-
DL-isocitrate
with Mn2+, pH 7.5, 37C
Microcystis aeruginosa
1.1.1.42
613
-
NADP+
with Mn2+, pH 7.5, 37C
Microcystis aeruginosa
1.1.1.42
787
-
NADP+
with Mg2+, pH 7.5, 37C
Microcystis aeruginosa