Bifunctional isocitrate-homoisocitrate dehydrogenase: a missing link in the evolution of beta-decarboxylating dehydrogenase
Miyazaki, K.; Biochem. Biophys. Res. Commun. 331, 341-346 (2005) 
Data extracted from this reference:
Cloned(Commentary)
EC Number
Commentary
Organism
1.1.1.41
expressed in Escherichia coli strain BL21-CodonPlus (DE3)-RIL
Pyrococcus horikoshii
1.1.1.87
DNA and amino acid sequence determination and analysis, expression of His-tagged enzyme in Escherichia coli
Pyrococcus horikoshii
1.1.1.286
-
Pyrococcus horikoshii
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.41
0.0164
-
isocitrate
6fold His-tagged enzyme, at 70°C in 50 mM N-2-hydroxyethylpiperadine-N0-2-ethanesulfonate-NaOH (pH 7.8), 0.2 M KCl, 0.2 mM MnCl2, 1mM NAD+
Pyrococcus horikoshii
1.1.1.41
0.0183
-
homoisocitrate
6fold His-tagged enzyme, at 70°C in 50 mM N-2-hydroxyethylpiperadine-N0-2-ethanesulfonate-NaOH (pH 7.8), 0.2 M KCl, 0.2 mM MnCl2, 1mM NAD+
Pyrococcus horikoshii
1.1.1.41
0.0771
-
NAD+
6fold His-tagged enzyme, at 70°C in 50 mM N-2-hydroxyethylpiperadine-N0-2-ethanesulfonate-NaOH (pH 7.8), 0.2 M KCl, 0.2 mM MnCl2
Pyrococcus horikoshii
1.1.1.87
0.0164
-
isocitrate
pH 7.8, 70°C, recombinant enzyme
Pyrococcus horikoshii
1.1.1.87
0.0183
-
homoisocitrate
pH 7.8, 70°C, recombinant enzyme
Pyrococcus horikoshii
1.1.1.286
0.0164
-
isocitrate
70°C, pH 7.8
Pyrococcus horikoshii
1.1.1.286
0.0183
-
homoisocitrate
70°C, pH 7.8
Pyrococcus horikoshii
1.1.1.286
0.0771
-
NAD+
70°C, pH 7.8
Pyrococcus horikoshii
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
1.1.1.41
K+
-
Pyrococcus horikoshii
1.1.1.41
Mn2+
-
Pyrococcus horikoshii
1.1.1.87
Mn2+
-
Pyrococcus horikoshii
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.1.1.87
3-isopropylmalate + NAD+
Pyrococcus horikoshii
-
?
-
-
?
1.1.1.87
homoisocitrate + NAD+
Pyrococcus horikoshii
-
?
-
-
?
1.1.1.87
isocitrate + NAD+
Pyrococcus horikoshii
-
?
-
-
?
1.1.1.87
additional information
Pyrococcus horikoshii
the enzyme is trifunctional performing the activities of 3-isopropylmalate, isocitrate, and homoisocitrate dehydrogenase in one pathway
?
-
-
-
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.1.1.41
Pyrococcus horikoshii
-
-
-
1.1.1.87
Pyrococcus horikoshii
A4CYJ9
gene PH1722 or hdh
-
1.1.1.286
Pyrococcus horikoshii
-
expression in Escherichia coli
-
Purification (Commentary)
EC Number
Commentary
Organism
1.1.1.41
Hi-Trap chelating HP column chromatography
Pyrococcus horikoshii
1.1.1.87
recombinnat His-tagged enzyme from Escherichia coli by metal affinity chromatography
Pyrococcus horikoshii
1.1.1.286
recombinant enzyme expressed in Escherichia coli, purification of inclusion bodies requires N-laurylsarcosine
Pyrococcus horikoshii
Renatured (Commentary)
EC Number
Commentary
Organism
1.1.1.41
using N-lauroylsarcosine to 0.3% v/v
Pyrococcus horikoshii
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
additional information
-
no activity on 3-isopropylmalate
Pyrococcus horikoshii
1.1.1.87
additional information
-
-
Pyrococcus horikoshii
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.41
D-homoisocitrate + NAD+
-
660907
Pyrococcus horikoshii
? + NADH
-
-
-
?
1.1.1.41
D-isocitrate + NAD+
-
660907
Pyrococcus horikoshii
2-oxoglutarate + CO2 + NADH
-
-
-
?
1.1.1.87
3-isopropylmalate + NAD+
-
660907
Pyrococcus horikoshii
?
-
-
-
?
1.1.1.87
homoisocitrate + NAD+
-
660907
Pyrococcus horikoshii
?
-
-
-
?
1.1.1.87
isocitrate + NAD+
-
660907
Pyrococcus horikoshii
?
-
-
-
?
1.1.1.87
additional information
the enzyme is trifunctional performing the activities of 3-isopropylmalate, isocitrate, and homoisocitrate dehydrogenase in one pathway
660907
Pyrococcus horikoshii
?
-
-
-
-
1.1.1.286
homoisocitrate + NAD+
-
660907
Pyrococcus horikoshii
? + NADH
-
-
-
?
1.1.1.286
isocitrate + NAD+
about 20times more efficient than homoisocitrate, NAD+ is preferred over NADP+
660907
Pyrococcus horikoshii
2-oxoglutarate + CO2 + NADH
-
-
-
?
1.1.1.286
isocitrate + NADP+
NAD+ is preferred over NADP+
660907
Pyrococcus horikoshii
2-oxoglutarate + CO2 + NADPH
-
-
-
?
1.1.1.286
additional information
no substrate: 3-isopropylmalate
660907
Pyrococcus horikoshii
?
-
-
-
-
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.1.1.87
70
-
assay at
Pyrococcus horikoshii
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.1.1.41
13.7
-
homoisocitrate
6fold His-tagged enzyme, at 70°C in 50 mM N-2-hydroxyethylpiperadine-N0-2-ethanesulfonate-NaOH (pH 7.8), 0.2 M KCl, 0.2 mM MnCl2, 1 mM NAD+
Pyrococcus horikoshii
1.1.1.41
14.8
-
isocitrate
6fold His-tagged enzyme, at 70°C in 50 mM N-2-hydroxyethylpiperadine-N0-2-ethanesulfonate-NaOH (pH 7.8), 0.2 M KCl, 0.2 mM MnCl2, 1 mM NAD+
Pyrococcus horikoshii
1.1.1.41
21.8
-
NAD+
6fold His-tagged enzyme, at 70°C in 50 mM N-2-hydroxyethylpiperadine-N0-2-ethanesulfonate-NaOH (pH 7.8), 0.2 M KCl, 0.2 mM MnCl2
Pyrococcus horikoshii
1.1.1.87
13.7
-
homoisocitrate
pH 7.8, 70°C, recombinant enzyme
Pyrococcus horikoshii
1.1.1.87
14.8
-
isocitrate
pH 7.8, 70°C, recombinant enzyme
Pyrococcus horikoshii
1.1.1.286
13.7
-
homoisocitrate
70°C, pH 7.8
Pyrococcus horikoshii
1.1.1.286
14.8
-
isocitrate
70°C, pH 7.8
Pyrococcus horikoshii
1.1.1.286
21.8
-
NAD+
70°C, pH 7.8
Pyrococcus horikoshii
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.87
7.8
-
assay at
Pyrococcus horikoshii
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.41
NAD+
1 mM
Pyrococcus horikoshii
1.1.1.87
NAD+
-
Pyrococcus horikoshii
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.41
expressed in Escherichia coli strain BL21-CodonPlus (DE3)-RIL
Pyrococcus horikoshii
1.1.1.87
DNA and amino acid sequence determination and analysis, expression of His-tagged enzyme in Escherichia coli
Pyrococcus horikoshii
1.1.1.286
-
Pyrococcus horikoshii
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.41
NAD+
1 mM
Pyrococcus horikoshii
1.1.1.87
NAD+
-
Pyrococcus horikoshii
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.41
0.0164
-
isocitrate
6fold His-tagged enzyme, at 70°C in 50 mM N-2-hydroxyethylpiperadine-N0-2-ethanesulfonate-NaOH (pH 7.8), 0.2 M KCl, 0.2 mM MnCl2, 1mM NAD+
Pyrococcus horikoshii
1.1.1.41
0.0183
-
homoisocitrate
6fold His-tagged enzyme, at 70°C in 50 mM N-2-hydroxyethylpiperadine-N0-2-ethanesulfonate-NaOH (pH 7.8), 0.2 M KCl, 0.2 mM MnCl2, 1mM NAD+
Pyrococcus horikoshii
1.1.1.41
0.0771
-
NAD+
6fold His-tagged enzyme, at 70°C in 50 mM N-2-hydroxyethylpiperadine-N0-2-ethanesulfonate-NaOH (pH 7.8), 0.2 M KCl, 0.2 mM MnCl2
Pyrococcus horikoshii
1.1.1.87
0.0164
-
isocitrate
pH 7.8, 70°C, recombinant enzyme
Pyrococcus horikoshii
1.1.1.87
0.0183
-
homoisocitrate
pH 7.8, 70°C, recombinant enzyme
Pyrococcus horikoshii
1.1.1.286
0.0164
-
isocitrate
70°C, pH 7.8
Pyrococcus horikoshii
1.1.1.286
0.0183
-
homoisocitrate
70°C, pH 7.8
Pyrococcus horikoshii
1.1.1.286
0.0771
-
NAD+
70°C, pH 7.8
Pyrococcus horikoshii
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
1.1.1.41
K+
-
Pyrococcus horikoshii
1.1.1.41
Mn2+
-
Pyrococcus horikoshii
1.1.1.87
Mn2+
-
Pyrococcus horikoshii
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.1.1.87
3-isopropylmalate + NAD+
Pyrococcus horikoshii
-
?
-
-
?
1.1.1.87
homoisocitrate + NAD+
Pyrococcus horikoshii
-
?
-
-
?
1.1.1.87
isocitrate + NAD+
Pyrococcus horikoshii
-
?
-
-
?
1.1.1.87
additional information
Pyrococcus horikoshii
the enzyme is trifunctional performing the activities of 3-isopropylmalate, isocitrate, and homoisocitrate dehydrogenase in one pathway
?
-
-
-
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.41
Hi-Trap chelating HP column chromatography
Pyrococcus horikoshii
1.1.1.87
recombinnat His-tagged enzyme from Escherichia coli by metal affinity chromatography
Pyrococcus horikoshii
1.1.1.286
recombinant enzyme expressed in Escherichia coli, purification of inclusion bodies requires N-laurylsarcosine
Pyrococcus horikoshii
Renatured (Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.41
using N-lauroylsarcosine to 0.3% v/v
Pyrococcus horikoshii
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
additional information
-
no activity on 3-isopropylmalate
Pyrococcus horikoshii
1.1.1.87
additional information
-
-
Pyrococcus horikoshii
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
D-homoisocitrate + NAD+
-
660907
Pyrococcus horikoshii
? + NADH
-
-
-
?
1.1.1.41
D-isocitrate + NAD+
-
660907
Pyrococcus horikoshii
2-oxoglutarate + CO2 + NADH
-
-
-
?
1.1.1.87
3-isopropylmalate + NAD+
-
660907
Pyrococcus horikoshii
?
-
-
-
?
1.1.1.87
homoisocitrate + NAD+
-
660907
Pyrococcus horikoshii
?
-
-
-
?
1.1.1.87
isocitrate + NAD+
-
660907
Pyrococcus horikoshii
?
-
-
-
?
1.1.1.87
additional information
the enzyme is trifunctional performing the activities of 3-isopropylmalate, isocitrate, and homoisocitrate dehydrogenase in one pathway
660907
Pyrococcus horikoshii
?
-
-
-
-
1.1.1.286
homoisocitrate + NAD+
-
660907
Pyrococcus horikoshii
? + NADH
-
-
-
?
1.1.1.286
isocitrate + NAD+
about 20times more efficient than homoisocitrate, NAD+ is preferred over NADP+
660907
Pyrococcus horikoshii
2-oxoglutarate + CO2 + NADH
-
-
-
?
1.1.1.286
isocitrate + NADP+
NAD+ is preferred over NADP+
660907
Pyrococcus horikoshii
2-oxoglutarate + CO2 + NADPH
-
-
-
?
1.1.1.286
additional information
no substrate: 3-isopropylmalate
660907
Pyrococcus horikoshii
?
-
-
-
-
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
1.1.1.87
70
-
assay at
Pyrococcus horikoshii
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
13.7
-
homoisocitrate
6fold His-tagged enzyme, at 70°C in 50 mM N-2-hydroxyethylpiperadine-N0-2-ethanesulfonate-NaOH (pH 7.8), 0.2 M KCl, 0.2 mM MnCl2, 1 mM NAD+
Pyrococcus horikoshii
1.1.1.41
14.8
-
isocitrate
6fold His-tagged enzyme, at 70°C in 50 mM N-2-hydroxyethylpiperadine-N0-2-ethanesulfonate-NaOH (pH 7.8), 0.2 M KCl, 0.2 mM MnCl2, 1 mM NAD+
Pyrococcus horikoshii
1.1.1.41
21.8
-
NAD+
6fold His-tagged enzyme, at 70°C in 50 mM N-2-hydroxyethylpiperadine-N0-2-ethanesulfonate-NaOH (pH 7.8), 0.2 M KCl, 0.2 mM MnCl2
Pyrococcus horikoshii
1.1.1.87
13.7
-
homoisocitrate
pH 7.8, 70°C, recombinant enzyme
Pyrococcus horikoshii
1.1.1.87
14.8
-
isocitrate
pH 7.8, 70°C, recombinant enzyme
Pyrococcus horikoshii
1.1.1.286
13.7
-
homoisocitrate
70°C, pH 7.8
Pyrococcus horikoshii
1.1.1.286
14.8
-
isocitrate
70°C, pH 7.8
Pyrococcus horikoshii
1.1.1.286
21.8
-
NAD+
70°C, pH 7.8
Pyrococcus horikoshii
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.87
7.8
-
assay at
Pyrococcus horikoshii