BRENDA - Enzyme Database

Biochemical and molecular characterization of the NAD(+)-dependent isocitrate dehydrogenase from the chemolithotroph Acidithiobacillus thiooxidans

Inoue, H.; Tamura, T.; Ehara, N.; Nishito, A.; Nakayama, Y.; Maekawa, M.; Imada, K.; Tanaka, H.; Inagaki, K.; FEMS Microbiol. Lett. 214, 127-132 (2002)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
1.1.1.41
gene icd, DNA and amino acid sequence determination and analysis, expression in enzyme-deficient Escherichia coli strain EB106
Acidithiobacillus thiooxidans
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.41
additional information
-
additional information
kinetic properties of the native and the recombinant enzyme are similar
Acidithiobacillus thiooxidans
1.1.1.41
0.18
-
NAD+
pH 9.0, 30°C, native enzyme
Acidithiobacillus thiooxidans
1.1.1.41
6.57
-
NADP+
pH 9.0, 30°C, native enzyme
Acidithiobacillus thiooxidans
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
1.1.1.41
Mg2+
-
Acidithiobacillus thiooxidans
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.1.1.41
45000
-
2 * 45000, SDS-PAGE
Acidithiobacillus thiooxidans
1.1.1.41
90000
-
gel filtration and native PAGE
Acidithiobacillus thiooxidans
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.1.1.41
isocitrate + NAD+
Acidithiobacillus thiooxidans
-
2-oxoglutarate + CO2 + NADH
-
-
?
1.1.1.41
isocitrate + NAD+
Acidithiobacillus thiooxidans ON107
-
2-oxoglutarate + CO2 + NADH
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.1.1.41
Acidithiobacillus thiooxidans
Q8GAX0
-
-
1.1.1.41
Acidithiobacillus thiooxidans ON107
Q8GAX0
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.1.1.41
multi-step purification of the native enzyme, 520fold to homogeneity, partial purification of the recombinant enzyme from Escherichia coli
Acidithiobacillus thiooxidans
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.1.1.41
120
-
purified native enzyme
Acidithiobacillus thiooxidans
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.41
isocitrate + NAD+
-
655607
Acidithiobacillus thiooxidans
2-oxoglutarate + CO2 + NADH
-
-
-
?
1.1.1.41
isocitrate + NAD+
-
655607
Acidithiobacillus thiooxidans ON107
2-oxoglutarate + CO2 + NADH
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
1.1.1.41
dimer
2 * 45000, SDS-PAGE
Acidithiobacillus thiooxidans
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.1.1.41
30
-
assay at
Acidithiobacillus thiooxidans
Temperature Stability [°C]
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
1.1.1.41
55
-
stable up to
Acidithiobacillus thiooxidans
1.1.1.41
70
-
inactivation after 30 min
Acidithiobacillus thiooxidans
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.1.1.41
21.3
-
NADP+
pH 9.0, 30°C, native enzyme
Acidithiobacillus thiooxidans
1.1.1.41
45.1
-
NAD+
pH 9.0, 30°C, native enzyme
Acidithiobacillus thiooxidans
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.41
8.5
-
-
Acidithiobacillus thiooxidans
pH Range
EC Number
pH Minimum
pH Maximum
Commentary
Organism
1.1.1.41
6
10.2
-
Acidithiobacillus thiooxidans
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.41
NAD+
highly preferred cofactor, 78fold higher activity than with NADP+
Acidithiobacillus thiooxidans
1.1.1.41
NADP+
78fold lower activity than with NAD+
Acidithiobacillus thiooxidans
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.41
gene icd, DNA and amino acid sequence determination and analysis, expression in enzyme-deficient Escherichia coli strain EB106
Acidithiobacillus thiooxidans
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.41
NAD+
highly preferred cofactor, 78fold higher activity than with NADP+
Acidithiobacillus thiooxidans
1.1.1.41
NADP+
78fold lower activity than with NAD+
Acidithiobacillus thiooxidans
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.41
additional information
-
additional information
kinetic properties of the native and the recombinant enzyme are similar
Acidithiobacillus thiooxidans
1.1.1.41
0.18
-
NAD+
pH 9.0, 30°C, native enzyme
Acidithiobacillus thiooxidans
1.1.1.41
6.57
-
NADP+
pH 9.0, 30°C, native enzyme
Acidithiobacillus thiooxidans
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
1.1.1.41
Mg2+
-
Acidithiobacillus thiooxidans
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.1.1.41
45000
-
2 * 45000, SDS-PAGE
Acidithiobacillus thiooxidans
1.1.1.41
90000
-
gel filtration and native PAGE
Acidithiobacillus thiooxidans
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.1.1.41
isocitrate + NAD+
Acidithiobacillus thiooxidans
-
2-oxoglutarate + CO2 + NADH
-
-
?
1.1.1.41
isocitrate + NAD+
Acidithiobacillus thiooxidans ON107
-
2-oxoglutarate + CO2 + NADH
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.41
multi-step purification of the native enzyme, 520fold to homogeneity, partial purification of the recombinant enzyme from Escherichia coli
Acidithiobacillus thiooxidans
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.1.1.41
120
-
purified native enzyme
Acidithiobacillus thiooxidans
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.41
isocitrate + NAD+
-
655607
Acidithiobacillus thiooxidans
2-oxoglutarate + CO2 + NADH
-
-
-
?
1.1.1.41
isocitrate + NAD+
-
655607
Acidithiobacillus thiooxidans ON107
2-oxoglutarate + CO2 + NADH
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.1.1.41
dimer
2 * 45000, SDS-PAGE
Acidithiobacillus thiooxidans
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.1.1.41
30
-
assay at
Acidithiobacillus thiooxidans
Temperature Stability [°C] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
1.1.1.41
55
-
stable up to
Acidithiobacillus thiooxidans
1.1.1.41
70
-
inactivation after 30 min
Acidithiobacillus thiooxidans
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.1.1.41
21.3
-
NADP+
pH 9.0, 30°C, native enzyme
Acidithiobacillus thiooxidans
1.1.1.41
45.1
-
NAD+
pH 9.0, 30°C, native enzyme
Acidithiobacillus thiooxidans
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.41
8.5
-
-
Acidithiobacillus thiooxidans
pH Range (protein specific)
EC Number
pH Minimum
pH Maximum
Commentary
Organism
1.1.1.41
6
10.2
-
Acidithiobacillus thiooxidans