BRENDA - Enzyme Database

NADP-dependent isocitrate dehydrogenase from the halophilic archaeon Haloferax volcanii: cloning, sequence determination and overexpression in Escherichia coli

Camacho, M.; Rodriguez-Arnedo, A.; Bonete, M.J.; FEMS Microbiol. Lett. 209, 155-160 (2002)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
1.1.1.42
IDCH gene, DNA and amino acid sequence determination by combinated PCR and screening of genomic lambda EMBL3 library, and sequence analysis, overexpression in Escherichia coli BL21(DE3) as insoluble protein in inclusion bodies
Haloferax volcanii
General Stability
EC Number
General Stability
Organism
1.1.1.42
3 M NaCl stabilizes the recombinant enzyme
Haloferax volcanii
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.42
0.093
-
NADP+
recombinant, renatured enzyme, pH 8.0
Haloferax volcanii
1.1.1.42
0.105
-
DL-isocitrate
recombinant, renatured enzyme, pH 8.0
Haloferax volcanii
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
1.1.1.42
Mg2+
-
Haloferax volcanii
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.1.1.42
45837
-
x * 45837, sequence calculation, x * 60000, recombinant, renatured enzyme, SDS-PAGE
Haloferax volcanii
1.1.1.42
60000
-
x * 45837, sequence calculation, x * 60000, recombinant, renatured enzyme, SDS-PAGE
Haloferax volcanii
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.1.1.42
Haloferax volcanii
Q8X277
halophilic archaeon
-
Purification (Commentary)
EC Number
Commentary
Organism
1.1.1.42
recombinant solubilized enzyme by ultrafiltration, and DEAE ion exchange chromatography, 6.4fold
Haloferax volcanii
Renatured (Commentary)
EC Number
Commentary
Organism
1.1.1.42
solubilization in presence of 3 M NaCl of recombinant enzyme from inclusion bodies after expression in Escherichia coli
Haloferax volcanii
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.1.1.42
23.7
-
purified recombinant enzyme
Haloferax volcanii
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+
-
655604
Haloferax volcanii
2-oxoglutarate + CO2 + NADPH
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
1.1.1.42
?
x * 45837, sequence calculation, x * 60000, recombinant, renatured enzyme, SDS-PAGE
Haloferax volcanii
Temperature Stability [°C]
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
1.1.1.42
60
-
recombinant enzyme, 93% remaining activity after 117 h, in presence of 3 M NaCl
Haloferax volcanii
1.1.1.42
80
-
recombinant enzyme, inactivation within 2.5 min, in presence of 3 M NaCl
Haloferax volcanii
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.42
8
-
assay at
Haloferax volcanii
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.42
NADP+
-
Haloferax volcanii
pI Value
EC Number
Organism
Commentary
pI Value Maximum
pI Value
1.1.1.42
Haloferax volcanii
calculated
-
4.9
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.42
IDCH gene, DNA and amino acid sequence determination by combinated PCR and screening of genomic lambda EMBL3 library, and sequence analysis, overexpression in Escherichia coli BL21(DE3) as insoluble protein in inclusion bodies
Haloferax volcanii
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.42
NADP+
-
Haloferax volcanii
General Stability (protein specific)
EC Number
General Stability
Organism
1.1.1.42
3 M NaCl stabilizes the recombinant enzyme
Haloferax volcanii
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.42
0.093
-
NADP+
recombinant, renatured enzyme, pH 8.0
Haloferax volcanii
1.1.1.42
0.105
-
DL-isocitrate
recombinant, renatured enzyme, pH 8.0
Haloferax volcanii
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
1.1.1.42
Mg2+
-
Haloferax volcanii
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.1.1.42
45837
-
x * 45837, sequence calculation, x * 60000, recombinant, renatured enzyme, SDS-PAGE
Haloferax volcanii
1.1.1.42
60000
-
x * 45837, sequence calculation, x * 60000, recombinant, renatured enzyme, SDS-PAGE
Haloferax volcanii
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.42
recombinant solubilized enzyme by ultrafiltration, and DEAE ion exchange chromatography, 6.4fold
Haloferax volcanii
Renatured (Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.42
solubilization in presence of 3 M NaCl of recombinant enzyme from inclusion bodies after expression in Escherichia coli
Haloferax volcanii
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.1.1.42
23.7
-
purified recombinant enzyme
Haloferax volcanii
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+
-
655604
Haloferax volcanii
2-oxoglutarate + CO2 + NADPH
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.1.1.42
?
x * 45837, sequence calculation, x * 60000, recombinant, renatured enzyme, SDS-PAGE
Haloferax volcanii
Temperature Stability [°C] (protein specific)
EC Number
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
1.1.1.42
60
-
recombinant enzyme, 93% remaining activity after 117 h, in presence of 3 M NaCl
Haloferax volcanii
1.1.1.42
80
-
recombinant enzyme, inactivation within 2.5 min, in presence of 3 M NaCl
Haloferax volcanii
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.42
8
-
assay at
Haloferax volcanii
pI Value (protein specific)
EC Number
Organism
Commentary
pI Value Maximum
pI Value
1.1.1.42
Haloferax volcanii
calculated
-
4.9