1.1.1.286
purified recombinant His6-tagged enzyme in apoform or in complex with substrates isocitrate, homoisocitrate, and 3-isopropylmalate, hanging drop vapour diffusion method, for the apoenzyme: mixing of 0.001 ml of 10 mg/ml protein in 20 mM Tris-HCl, pH 8.0, and 150 mM NaCl, with 0.001 ml of reservoir solution containing 100 mM imidazole-HCl, pH 6.5, and 2.3 M NaCl, equilibration against 1 m of reservoir solution, at 20°C, for the ternary complex with homoisocitrate and Mn2+: mixing of 0.0015 ml of 10 mg/ml protein in 20 mM Tris-HCl, pH 8.0, 150 mM NaCl, 10% glycerol, 1 mM MnSO4, and 5 mM homoisocitrate, with 0.0015 ml of reservoir solution containing 100 mM HEPES-NaOH, pH 7.5, 200 mM NaCl, and 10% v/v 2-propanol, at 20°C, for the ternary complex with homoisocitrate and Mn2+: mixing of 0.0015 ml of 5 mg/ml protein in 20 mM Tris-HCl, pH 8.0, 150 mM NaCl, 10% glycerol, 1 mM MnSO4, and 5 mM isocitrate, with 0.0015 ml of reservoir solution containing 100 mM TrisHCl, pH 7.5, 200 mM NaCl, and 30% v/v 2-methyl-2,4-pentanediol, at 20°C, and for the ternary complex with 3-isopropylmalate and Mn2+: mixing of 0.001 ml of 15 mg/ml protein in 20 mM Tris-HCl, pH 8.0, 150 mM NaCl, 10% glycerol, 1 mM MnSO4, and 5 mM 3-isopropylmalate, with 0.001 ml of reservoir solution containing 100 mM HEPES-NaOH, pH 7.5, 200 mM NaCl, and 36% v/v 2-methyl-2,4-pentanediol, at 20°C, X-ray diffraction structure determination and analysis at 1.7 A resolution for the apoenzyme, and at 2.50-2.64 A resolution for the complexed enzyme
Thermococcus kodakarensis
1.1.1.286
I82N
site-directed mutagenesis, the mutant shows reduced activity and altered substrate specificity compared to the wild-type enzyme
Thermococcus kodakarensis
1.1.1.286
L81P
site-directed mutagenesis, the mutant shows reduced activity and altered substrate specificity compared to the wild-type enzyme
Thermococcus kodakarensis
1.1.1.286
L83R
site-directed mutagenesis, the mutant shows reduced activity and altered substrate specificity compared to the wild-type enzyme
Thermococcus kodakarensis
1.1.1.286
additional information
generation of a gene-disruption TK0280 deletion mutan strain, functional complementation by expression of the gene icdh from Thermus thermophilus strain Hb27, but not by genes hicdh and leuB from this organism
Thermococcus kodakarensis
1.1.1.286
S80A
site-directed mutagenesis, the mutant shows reduced activity and altered substrate specificity compared to the wild-type enzyme
Thermococcus kodakarensis
1.1.1.286
T71V
site-directed mutagenesis, the mutant shows reduced activity and altered substrate specificity compared to the wild-type enzyme
Thermococcus kodakarensis
1.1.1.286
additional information
additional information
Michaelis-Menten kinetics
Thermococcus kodakarensis
1.1.1.286
0.0073
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme
Thermococcus kodakarensis
1.1.1.286
0.014
isocitrate
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme
Thermococcus kodakarensis
1.1.1.286
0.036
NAD+
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme, with homoisocitrate
Thermococcus kodakarensis
1.1.1.286
0.037
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant S80A
Thermococcus kodakarensis
1.1.1.286
0.042
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant L81P
Thermococcus kodakarensis
1.1.1.286
0.05
3-isopropylmalate
pH 8.0, 60°C, recombinant His6-tagged mutant S80A
Thermococcus kodakarensis
1.1.1.286
0.1
NAD+
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme, with isocitrate
Thermococcus kodakarensis
1.1.1.286
0.16
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant L83R, with homoisocitrate
Thermococcus kodakarensis
1.1.1.286
0.17
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant L83R
Thermococcus kodakarensis
1.1.1.286
0.25
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant T71V, with isocitrate
Thermococcus kodakarensis
1.1.1.286
0.3
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant S80A
Thermococcus kodakarensis
1.1.1.286
0.31
3-isopropylmalate
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme
Thermococcus kodakarensis
1.1.1.286
0.32
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant L81P, with homoisocitrate
Thermococcus kodakarensis
1.1.1.286
0.46
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant I82N
Thermococcus kodakarensis
1.1.1.286
0.5
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant T71V
Thermococcus kodakarensis
1.1.1.286
0.58
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant L83R
Thermococcus kodakarensis
1.1.1.286
0.6
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant L81P, with isocitrate
Thermococcus kodakarensis
1.1.1.286
0.62
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant L83R, with isocitrate
Thermococcus kodakarensis
1.1.1.286
0.64
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant I82N, with homoisocitrate
Thermococcus kodakarensis
1.1.1.286
0.68
3-isopropylmalate
pH 8.0, 60°C, recombinant His6-tagged mutant L81P
Thermococcus kodakarensis
1.1.1.286
0.71
3-isopropylmalate
pH 8.0, 60°C, recombinant His6-tagged mutant T71V
Thermococcus kodakarensis
1.1.1.286
0.76
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant S80A, with isocitrate
Thermococcus kodakarensis
1.1.1.286
0.92
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant S80A, with homoisocitrate
Thermococcus kodakarensis
1.1.1.286
0.94
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant T71V
Thermococcus kodakarensis
1.1.1.286
1
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant T71V, with 3-isopropylmalate
Thermococcus kodakarensis
1.1.1.286
1.2
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant L81P
Thermococcus kodakarensis
1.1.1.286
1.2
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant S80A, with 3-isopropylmalate
Thermococcus kodakarensis
1.1.1.286
1.3
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant L81P, with 3-isopropylmalate
Thermococcus kodakarensis
1.1.1.286
1.5
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant T71V, with homoisocitrate
Thermococcus kodakarensis
1.1.1.286
1.5
NAD+
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme, with 3-isopropylmalate
Thermococcus kodakarensis
1.1.1.286
1.8
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant I82N, with isocitrate
Thermococcus kodakarensis
1.1.1.286
2.2
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant I82N
Thermococcus kodakarensis
1.1.1.286
3-isopropylmalate + NAD+
740021
Thermococcus kodakarensis
2-oxoisocaproate + CO2 + NADH + H+
?
1.1.1.286
3-isopropylmalate + NAD+
740021
Thermococcus kodakarensis KRD1
2-oxoisocaproate + CO2 + NADH + H+
?
1.1.1.286
homoisocitrate + NAD+
740021
Thermococcus kodakarensis
2-oxoadipate + CO2 + NADH + H+
?
1.1.1.286
homoisocitrate + NAD+
740021
Thermococcus kodakarensis KRD1
2-oxoadipate + CO2 + NADH + H+
?
1.1.1.286
isocitrate + NAD+
740021
Thermococcus kodakarensis
2-oxoglutarate + CO2 + NADH
?
1.1.1.286
isocitrate + NAD+
740021
Thermococcus kodakarensis KRD1
2-oxoglutarate + CO2 + NADH
?
1.1.1.286
additional information
residues Thr71 and Ser80 play important roles in the recognition of homoisocitrate and isocitrate while the hydrophobic region consisting of Ile82 and Leu83 is responsible for the recognition of 3-isopropylmalate. Importance of a water-mediated hydrogen bond network for the stabilization of the beta3-alpha4 loop, including the Thr71 residue, with respect to the promiscuity of the substrate specificity of TK0280
740021
Thermococcus kodakarensis
?
?
1.1.1.286
additional information
residues Thr71 and Ser80 play important roles in the recognition of homoisocitrate and isocitrate while the hydrophobic region consisting of Ile82 and Leu83 is responsible for the recognition of 3-isopropylmalate. Importance of a water-mediated hydrogen bond network for the stabilization of the beta3-alpha4 loop, including the Thr71 residue, with respect to the promiscuity of the substrate specificity of TK0280
740021
Thermococcus kodakarensis KRD1
?
?
1.1.1.286
0.22
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant T71V
Thermococcus kodakarensis
1.1.1.286
0.71
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant S80A
Thermococcus kodakarensis
1.1.1.286
1
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant I82N
Thermococcus kodakarensis
1.1.1.286
1.1
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant L81P
Thermococcus kodakarensis
1.1.1.286
1.5
3-isopropylmalate
pH 8.0, 60°C, recombinant His6-tagged mutant L81P
Thermococcus kodakarensis
1.1.1.286
2.3
3-isopropylmalate
pH 8.0, 60°C, recombinant His6-tagged mutant S80A
Thermococcus kodakarensis
1.1.1.286
4.8
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant T71V
Thermococcus kodakarensis
1.1.1.286
4.8
3-isopropylmalate
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme
Thermococcus kodakarensis
1.1.1.286
5
isocitrate
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme
Thermococcus kodakarensis
1.1.1.286
6.6
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant L83R
Thermococcus kodakarensis
1.1.1.286
6.9
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant S80A
Thermococcus kodakarensis
1.1.1.286
9.4
3-isopropylmalate
pH 8.0, 60°C, recombinant His6-tagged mutant T71V
Thermococcus kodakarensis
1.1.1.286
13
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant L81P
Thermococcus kodakarensis
1.1.1.286
17
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme
Thermococcus kodakarensis
1.1.1.286
43
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant L83R
Thermococcus kodakarensis
1.1.1.286
45
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant I82N
Thermococcus kodakarensis
1.1.1.286
purified recombinant His6-tagged enzyme in apoform or in complex with substrates isocitrate, homoisocitrate, and 3-isopropylmalate, hanging drop vapour diffusion method, for the apoenzyme: mixing of 0.001 ml of 10 mg/ml protein in 20 mM Tris-HCl, pH 8.0, and 150 mM NaCl, with 0.001 ml of reservoir solution containing 100 mM imidazole-HCl, pH 6.5, and 2.3 M NaCl, equilibration against 1 m of reservoir solution, at 20°C, for the ternary complex with homoisocitrate and Mn2+: mixing of 0.0015 ml of 10 mg/ml protein in 20 mM Tris-HCl, pH 8.0, 150 mM NaCl, 10% glycerol, 1 mM MnSO4, and 5 mM homoisocitrate, with 0.0015 ml of reservoir solution containing 100 mM HEPES-NaOH, pH 7.5, 200 mM NaCl, and 10% v/v 2-propanol, at 20°C, for the ternary complex with homoisocitrate and Mn2+: mixing of 0.0015 ml of 5 mg/ml protein in 20 mM Tris-HCl, pH 8.0, 150 mM NaCl, 10% glycerol, 1 mM MnSO4, and 5 mM isocitrate, with 0.0015 ml of reservoir solution containing 100 mM TrisHCl, pH 7.5, 200 mM NaCl, and 30% v/v 2-methyl-2,4-pentanediol, at 20°C, and for the ternary complex with 3-isopropylmalate and Mn2+: mixing of 0.001 ml of 15 mg/ml protein in 20 mM Tris-HCl, pH 8.0, 150 mM NaCl, 10% glycerol, 1 mM MnSO4, and 5 mM 3-isopropylmalate, with 0.001 ml of reservoir solution containing 100 mM HEPES-NaOH, pH 7.5, 200 mM NaCl, and 36% v/v 2-methyl-2,4-pentanediol, at 20°C, X-ray diffraction structure determination and analysis at 1.7 A resolution for the apoenzyme, and at 2.50-2.64 A resolution for the complexed enzyme
Thermococcus kodakarensis
1.1.1.286
I82N
site-directed mutagenesis, the mutant shows reduced activity and altered substrate specificity compared to the wild-type enzyme
Thermococcus kodakarensis
1.1.1.286
L81P
site-directed mutagenesis, the mutant shows reduced activity and altered substrate specificity compared to the wild-type enzyme
Thermococcus kodakarensis
1.1.1.286
L83R
site-directed mutagenesis, the mutant shows reduced activity and altered substrate specificity compared to the wild-type enzyme
Thermococcus kodakarensis
1.1.1.286
additional information
generation of a gene-disruption TK0280 deletion mutan strain, functional complementation by expression of the gene icdh from Thermus thermophilus strain Hb27, but not by genes hicdh and leuB from this organism
Thermococcus kodakarensis
1.1.1.286
S80A
site-directed mutagenesis, the mutant shows reduced activity and altered substrate specificity compared to the wild-type enzyme
Thermococcus kodakarensis
1.1.1.286
T71V
site-directed mutagenesis, the mutant shows reduced activity and altered substrate specificity compared to the wild-type enzyme
Thermococcus kodakarensis
1.1.1.286
additional information
additional information
Michaelis-Menten kinetics
Thermococcus kodakarensis
1.1.1.286
0.0073
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme
Thermococcus kodakarensis
1.1.1.286
0.014
isocitrate
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme
Thermococcus kodakarensis
1.1.1.286
0.036
NAD+
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme, with homoisocitrate
Thermococcus kodakarensis
1.1.1.286
0.037
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant S80A
Thermococcus kodakarensis
1.1.1.286
0.042
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant L81P
Thermococcus kodakarensis
1.1.1.286
0.05
3-isopropylmalate
pH 8.0, 60°C, recombinant His6-tagged mutant S80A
Thermococcus kodakarensis
1.1.1.286
0.1
NAD+
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme, with isocitrate
Thermococcus kodakarensis
1.1.1.286
0.16
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant L83R, with homoisocitrate
Thermococcus kodakarensis
1.1.1.286
0.17
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant L83R
Thermococcus kodakarensis
1.1.1.286
0.25
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant T71V, with isocitrate
Thermococcus kodakarensis
1.1.1.286
0.3
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant S80A
Thermococcus kodakarensis
1.1.1.286
0.31
3-isopropylmalate
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme
Thermococcus kodakarensis
1.1.1.286
0.32
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant L81P, with homoisocitrate
Thermococcus kodakarensis
1.1.1.286
0.46
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant I82N
Thermococcus kodakarensis
1.1.1.286
0.5
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant T71V
Thermococcus kodakarensis
1.1.1.286
0.58
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant L83R
Thermococcus kodakarensis
1.1.1.286
0.6
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant L81P, with isocitrate
Thermococcus kodakarensis
1.1.1.286
0.62
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant L83R, with isocitrate
Thermococcus kodakarensis
1.1.1.286
0.64
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant I82N, with homoisocitrate
Thermococcus kodakarensis
1.1.1.286
0.68
3-isopropylmalate
pH 8.0, 60°C, recombinant His6-tagged mutant L81P
Thermococcus kodakarensis
1.1.1.286
0.71
3-isopropylmalate
pH 8.0, 60°C, recombinant His6-tagged mutant T71V
Thermococcus kodakarensis
1.1.1.286
0.76
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant S80A, with isocitrate
Thermococcus kodakarensis
1.1.1.286
0.92
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant S80A, with homoisocitrate
Thermococcus kodakarensis
1.1.1.286
0.94
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant T71V
Thermococcus kodakarensis
1.1.1.286
1
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant T71V, with 3-isopropylmalate
Thermococcus kodakarensis
1.1.1.286
1.2
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant L81P
Thermococcus kodakarensis
1.1.1.286
1.2
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant S80A, with 3-isopropylmalate
Thermococcus kodakarensis
1.1.1.286
1.3
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant L81P, with 3-isopropylmalate
Thermococcus kodakarensis
1.1.1.286
1.5
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant T71V, with homoisocitrate
Thermococcus kodakarensis
1.1.1.286
1.5
NAD+
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme, with 3-isopropylmalate
Thermococcus kodakarensis
1.1.1.286
1.8
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant I82N, with isocitrate
Thermococcus kodakarensis
1.1.1.286
2.2
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant I82N
Thermococcus kodakarensis
1.1.1.286
3-isopropylmalate + NAD+
740021
Thermococcus kodakarensis
2-oxoisocaproate + CO2 + NADH + H+
?
1.1.1.286
3-isopropylmalate + NAD+
740021
Thermococcus kodakarensis KRD1
2-oxoisocaproate + CO2 + NADH + H+
?
1.1.1.286
homoisocitrate + NAD+
740021
Thermococcus kodakarensis
2-oxoadipate + CO2 + NADH + H+
?
1.1.1.286
homoisocitrate + NAD+
740021
Thermococcus kodakarensis KRD1
2-oxoadipate + CO2 + NADH + H+
?
1.1.1.286
isocitrate + NAD+
740021
Thermococcus kodakarensis
2-oxoglutarate + CO2 + NADH
?
1.1.1.286
isocitrate + NAD+
740021
Thermococcus kodakarensis KRD1
2-oxoglutarate + CO2 + NADH
?
1.1.1.286
additional information
residues Thr71 and Ser80 play important roles in the recognition of homoisocitrate and isocitrate while the hydrophobic region consisting of Ile82 and Leu83 is responsible for the recognition of 3-isopropylmalate. Importance of a water-mediated hydrogen bond network for the stabilization of the beta3-alpha4 loop, including the Thr71 residue, with respect to the promiscuity of the substrate specificity of TK0280
740021
Thermococcus kodakarensis
?
?
1.1.1.286
additional information
residues Thr71 and Ser80 play important roles in the recognition of homoisocitrate and isocitrate while the hydrophobic region consisting of Ile82 and Leu83 is responsible for the recognition of 3-isopropylmalate. Importance of a water-mediated hydrogen bond network for the stabilization of the beta3-alpha4 loop, including the Thr71 residue, with respect to the promiscuity of the substrate specificity of TK0280
740021
Thermococcus kodakarensis KRD1
?
?
1.1.1.286
0.22
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant T71V
Thermococcus kodakarensis
1.1.1.286
0.71
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant S80A
Thermococcus kodakarensis
1.1.1.286
1
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant I82N
Thermococcus kodakarensis
1.1.1.286
1.1
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant L81P
Thermococcus kodakarensis
1.1.1.286
1.5
3-isopropylmalate
pH 8.0, 60°C, recombinant His6-tagged mutant L81P
Thermococcus kodakarensis
1.1.1.286
2.3
3-isopropylmalate
pH 8.0, 60°C, recombinant His6-tagged mutant S80A
Thermococcus kodakarensis
1.1.1.286
4.8
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant T71V
Thermococcus kodakarensis
1.1.1.286
4.8
3-isopropylmalate
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme
Thermococcus kodakarensis
1.1.1.286
5
isocitrate
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme
Thermococcus kodakarensis
1.1.1.286
6.6
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant L83R
Thermococcus kodakarensis
1.1.1.286
6.9
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant S80A
Thermococcus kodakarensis
1.1.1.286
9.4
3-isopropylmalate
pH 8.0, 60°C, recombinant His6-tagged mutant T71V
Thermococcus kodakarensis
1.1.1.286
13
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant L81P
Thermococcus kodakarensis
1.1.1.286
17
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme
Thermococcus kodakarensis
1.1.1.286
43
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant L83R
Thermococcus kodakarensis
1.1.1.286
45
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant I82N
Thermococcus kodakarensis
1.1.1.286
0.44
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant T71V
Thermococcus kodakarensis
1.1.1.286
0.45
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant I82N
Thermococcus kodakarensis
1.1.1.286
0.56
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant I82N, with isocitrate
Thermococcus kodakarensis
1.1.1.286
0.88
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant T71V, with isocitrate
Thermococcus kodakarensis
1.1.1.286
0.92
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant L81P
Thermococcus kodakarensis
1.1.1.286
0.93
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant S80A, with isocitrate
Thermococcus kodakarensis
1.1.1.286
1.2
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant L81P, with 3-isopropylmalate
Thermococcus kodakarensis
1.1.1.286
1.8
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant L81P, with isocitrate
Thermococcus kodakarensis
1.1.1.286
1.9
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant S80A, with 3-isopropylmalate
Thermococcus kodakarensis
1.1.1.286
2.2
3-isopropylmalate
pH 8.0, 60°C, recombinant His6-tagged mutant L81P
Thermococcus kodakarensis
1.1.1.286
2.4
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant S80A
Thermococcus kodakarensis
1.1.1.286
3.2
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant T71V, with homoisocitrate
Thermococcus kodakarensis
1.1.1.286
3.2
NAD+
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme, with 3-isopropylmalate
Thermococcus kodakarensis
1.1.1.286
5.3
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant T71V
Thermococcus kodakarensis
1.1.1.286
7.5
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant S80A, with homoisocitrate
Thermococcus kodakarensis
1.1.1.286
9.4
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant T71V, with 3-isopropylmalate
Thermococcus kodakarensis
1.1.1.286
11
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant L83R
Thermococcus kodakarensis
1.1.1.286
11
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant L83R, with isocitrate
Thermococcus kodakarensis
1.1.1.286
13
3-isopropylmalate
pH 8.0, 60°C, recombinant His6-tagged mutant T71V
Thermococcus kodakarensis
1.1.1.286
15
3-isopropylmalate
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme
Thermococcus kodakarensis
1.1.1.286
41
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant L81P, with homoisocitrate
Thermococcus kodakarensis
1.1.1.286
46
3-isopropylmalate
pH 8.0, 60°C, recombinant His6-tagged mutant S80A
Thermococcus kodakarensis
1.1.1.286
50
NAD+
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme, with isocitrate
Thermococcus kodakarensis
1.1.1.286
70
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant I82N, with homoisocitrate
Thermococcus kodakarensis
1.1.1.286
98
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant I82N
Thermococcus kodakarensis
1.1.1.286
190
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant S80A
Thermococcus kodakarensis
1.1.1.286
250
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant L83R
Thermococcus kodakarensis
1.1.1.286
270
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant L83R, with homoisocitrate
Thermococcus kodakarensis
1.1.1.286
310
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant L81P
Thermococcus kodakarensis
1.1.1.286
360
isocitrate
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme
Thermococcus kodakarensis
1.1.1.286
470
NAD+
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme, with homoisocitrate
Thermococcus kodakarensis
1.1.1.286
2300
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme
Thermococcus kodakarensis
1.1.1.286
0.44
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant T71V
Thermococcus kodakarensis
1.1.1.286
0.45
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant I82N
Thermococcus kodakarensis
1.1.1.286
0.56
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant I82N, with isocitrate
Thermococcus kodakarensis
1.1.1.286
0.88
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant T71V, with isocitrate
Thermococcus kodakarensis
1.1.1.286
0.92
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant L81P
Thermococcus kodakarensis
1.1.1.286
0.93
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant S80A, with isocitrate
Thermococcus kodakarensis
1.1.1.286
1.2
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant L81P, with 3-isopropylmalate
Thermococcus kodakarensis
1.1.1.286
1.8
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant L81P, with isocitrate
Thermococcus kodakarensis
1.1.1.286
1.9
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant S80A, with 3-isopropylmalate
Thermococcus kodakarensis
1.1.1.286
2.2
3-isopropylmalate
pH 8.0, 60°C, recombinant His6-tagged mutant L81P
Thermococcus kodakarensis
1.1.1.286
2.4
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant S80A
Thermococcus kodakarensis
1.1.1.286
3.2
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant T71V, with homoisocitrate
Thermococcus kodakarensis
1.1.1.286
3.2
NAD+
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme, with 3-isopropylmalate
Thermococcus kodakarensis
1.1.1.286
5.3
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant T71V
Thermococcus kodakarensis
1.1.1.286
7.5
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant S80A, with homoisocitrate
Thermococcus kodakarensis
1.1.1.286
9.4
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant T71V, with 3-isopropylmalate
Thermococcus kodakarensis
1.1.1.286
11
isocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant L83R
Thermococcus kodakarensis
1.1.1.286
11
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant L83R, with isocitrate
Thermococcus kodakarensis
1.1.1.286
13
3-isopropylmalate
pH 8.0, 60°C, recombinant His6-tagged mutant T71V
Thermococcus kodakarensis
1.1.1.286
15
3-isopropylmalate
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme
Thermococcus kodakarensis
1.1.1.286
41
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant L81P, with homoisocitrate
Thermococcus kodakarensis
1.1.1.286
46
3-isopropylmalate
pH 8.0, 60°C, recombinant His6-tagged mutant S80A
Thermococcus kodakarensis
1.1.1.286
50
NAD+
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme, with isocitrate
Thermococcus kodakarensis
1.1.1.286
70
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant I82N, with homoisocitrate
Thermococcus kodakarensis
1.1.1.286
98
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant I82N
Thermococcus kodakarensis
1.1.1.286
190
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant S80A
Thermococcus kodakarensis
1.1.1.286
250
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant L83R
Thermococcus kodakarensis
1.1.1.286
270
NAD+
pH 8.0, 60°C, recombinant His6-tagged mutant L83R, with homoisocitrate
Thermococcus kodakarensis
1.1.1.286
310
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged mutant L81P
Thermococcus kodakarensis
1.1.1.286
360
isocitrate
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme
Thermococcus kodakarensis
1.1.1.286
470
NAD+
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme, with homoisocitrate
Thermococcus kodakarensis
1.1.1.286
2300
homoisocitrate
pH 8.0, 60°C, recombinant His6-tagged wild-type enzyme
Thermococcus kodakarensis