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Literature summary extracted from

  • Watanabe, K.; Yamagishi, A.
    The effects of multiple ancestral residues on the Thermus thermophilus 3-isopropylmalate dehydrogenase (2006), FEBS Lett., 580, 3867-3871.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.85 gene leuB, expression of wild-type and mutants in Escherichia coli strain MA153 Thermus thermophilus

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.85 A335E site-directed mutagenesis, exchange of a residue for that of ancestral mutants, the mutant shows increased thermal stability compared to the wild-type enzyme, the mutant shows increased thermal stability compared to the wild-type enzyme Thermus thermophilus
1.1.1.85 D184H site-directed mutagenesis, exchange of a residue for that of ancestral mutants, the mutant shows increased catalytic efficiency with NAD+ compared to the wild-type enzyme Thermus thermophilus
1.1.1.85 F53L site-directed mutagenesis, exchange of a residue for that of ancestral mutants, the mutant shows increased catalytic efficiency with NAD+ compared to the wild-type enzyme Thermus thermophilus
1.1.1.85 H179K site-directed mutagenesis, exchange of a residue for that of ancestral mutants, the mutant shows increased catalytic efficiency with NAD+ compared to the wild-type enzyme Thermus thermophilus
1.1.1.85 L134N site-directed mutagenesis, exchange of a residue for that of ancestral mutants, the mutant shows increased thermal stability compared to the wild-type enzyme, the mutant shows highly increased thermal stability compared to the wild-type enzyme Thermus thermophilus
1.1.1.85 L134N/V181T/P324T/A335E site-directed mutagenesis, exchange of residues for those of ancestral mutants, the mutant shows increased thermal stability compared to the wild-type enzyme, the mutant shows increased thermal stability compared to the wild-type enzyme Thermus thermophilus
1.1.1.85 P324T site-directed mutagenesis, exchange of a residue for that of ancestral mutants, the mutant shows increased thermal stability compared to the wild-type enzyme, the mutant shows increased thermal stability compared to the wild-type enzyme Thermus thermophilus
1.1.1.85 P56E site-directed mutagenesis, exchange of a residue for that of ancestral mutants, the mutant shows increased catalytic efficiency with NAD+ compared to the wild-type enzyme Thermus thermophilus
1.1.1.85 R58L site-directed mutagenesis, exchange of a residue for that of ancestral mutants, the mutant shows increased catalytic efficiency with NAD+ compared to the wild-type enzyme Thermus thermophilus
1.1.1.85 S261N site-directed mutagenesis, exchange of a residue for that of ancestral mutants, the mutant shows slightly increased catalytic efficiency with NAD+ compared to the wild-type enzyme Thermus thermophilus
1.1.1.85 V181T site-directed mutagenesis, exchange of a residue for that of ancestral mutants, the mutant shows decreased thermal stability compared to the wild-type enzyme, the mutant shows decreased thermal stability compared to the wild-type enzyme Thermus thermophilus
1.1.1.85 V181T/P324T/A335E site-directed mutagenesis, exchange of residues for those of ancestral mutants, the mutant highly shows increased thermal stability compared to the wild-type enzyme, the mutant shows decreased thermal stability compared to the wild-type enzyme Thermus thermophilus
1.1.1.85 V61I site-directed mutagenesis, exchange of a residue for that of ancestral mutants, the mutant shows slightly increased catalytic efficiency with NAD+ compared to the wild-type enzyme Thermus thermophilus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.85 0.0061
-
 threo-D,L-isopropylmalate pH 7.6, 70°C, recombinant wild-type enzyme Thermus thermophilus
1.1.1.85 0.0063
-
 threo-D,L-isopropylmalate pH 7.6, 70°C, recombinant mutant V181T/P324T/A335E Thermus thermophilus
1.1.1.85 0.007
-
 threo-D,L-isopropylmalate pH 7.6, 70°C, recombinant mutant L134N Thermus thermophilus
1.1.1.85 0.0161
-
 threo-D,L-isopropylmalate pH 7.6, 70°C, recombinant mutant L134N/V181T/P324T/A335E Thermus thermophilus
1.1.1.85 0.172
-
 NAD+ pH 7.6, 70°C, recombinant mutant L134N Thermus thermophilus
1.1.1.85 0.255
-
 NAD+ pH 7.6, 70°C, recombinant mutant L134N/V181T/P324T/A335E Thermus thermophilus
1.1.1.85 0.336
-
 NAD+ pH 7.6, 70°C, recombinant mutant H197K Thermus thermophilus
1.1.1.85 0.359
-
 NAD+ pH 7.6, 70°C, recombinant mutant A335E Thermus thermophilus
1.1.1.85 0.371
-
 NAD+ pH 7.6, 70°C, recombinant mutant P324T Thermus thermophilus
1.1.1.85 0.401
-
 NAD+ pH 7.6, 70°C, recombinant mutant R58L Thermus thermophilus
1.1.1.85 0.428
-
 NAD+ pH 7.6, 70°C, recombinant mutant P56E Thermus thermophilus
1.1.1.85 0.468
-
 NAD+ pH 7.6, 70°C, recombinant mutant F53L Thermus thermophilus
1.1.1.85 0.514
-
 NAD+ pH 7.6, 70°C, recombinant wild-type enzyme Thermus thermophilus
1.1.1.85 0.52
-
 NAD+ pH 7.6, 70°C, recombinant mutant V61I Thermus thermophilus
1.1.1.85 0.567
-
 NAD+ pH 7.6, 70°C, recombinant mutant V181T/P324T/A335E Thermus thermophilus
1.1.1.85 0.625
-
 NAD+ pH 7.6, 70°C, recombinant mutant S261N Thermus thermophilus
1.1.1.85 0.642
-
 NAD+ pH 7.6, 70°C, recombinant mutant V181T Thermus thermophilus
1.1.1.85 0.673
-
 NAD+ pH 7.6, 70°C, recombinant mutant D184H Thermus thermophilus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.85 Mg2+
-
 Thermus thermophilus

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.85 Thermus thermophilus Q5SIY4 gene leuB
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.85 recombinant wild-type and mutants from Escherichia coli strain MA153 by anion exchange and adsorption chromatography Thermus thermophilus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.85 threo-DL-isopropylmalate + NAD+
-
 Thermus thermophilus 2-isopropyl-3-oxosuccinate + NADH
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.1.1.85 IPMDH
-
 Thermus thermophilus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.85 70
-
 assay at Thermus thermophilus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.85 50
-
 thermal stability curves of wild-type and mutant enzymes at pH 7.6 in presence of 0.5 mM EDTA, overview Thermus thermophilus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.85 10.2
-
 NAD+ pH 7.6, 70°C, recombinant mutant V181T Thermus thermophilus
1.1.1.85 11.2
-
 NAD+ pH 7.6, 70°C, recombinant mutant D184H Thermus thermophilus
1.1.1.85 13.2
-
 NAD+ pH 7.6, 70°C, recombinant mutant V181T/P324T/A335E Thermus thermophilus
1.1.1.85 13.4
-
 NAD+ pH 7.6, 70°C, recombinant wild-type enzyme Thermus thermophilus
1.1.1.85 13.8
-
 NAD+ pH 7.6, 70°C, recombinant mutant P324T Thermus thermophilus
1.1.1.85 14.6
-
 NAD+ pH 7.6, 70°C, recombinant mutant P56E Thermus thermophilus
1.1.1.85 15
-
 NAD+ pH 7.6, 70°C, recombinant mutants F53L and V61I Thermus thermophilus
1.1.1.85 17.7
-
 NAD+ pH 7.6, 70°C, recombinant mutant R58L Thermus thermophilus
1.1.1.85 18.2
-
 NAD+ pH 7.6, 70°C, recombinant mutants S261N and A335E Thermus thermophilus
1.1.1.85 21.3
-
 NAD+ pH 7.6, 70°C, recombinant mutant H197K Thermus thermophilus
1.1.1.85 25.2
-
 NAD+ pH 7.6, 70°C, recombinant mutant L134N/V181T/P324T/A335E Thermus thermophilus
1.1.1.85 29.9
-
 NAD+ pH 7.6, 70°C, recombinant mutant L134N Thermus thermophilus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.85 7.6
-
 assay at Thermus thermophilus

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.85 NAD+
-
 Thermus thermophilus