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Literature summary for 1.1.1.85 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
Search for more literature for 1.1.1.85

Cloned(Commentary)

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

Protein Variants

Protein Variants Comment Organism
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
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
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
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
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
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
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
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
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
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
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
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
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]

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

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+
-
 Thermus thermophilus

Organism

Organism UniProt Comment Textmining
Thermus thermophilus Q5SIY4 gene leuB
-

Purification (Commentary)

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

Substrates and Products (Substrate)

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

Synonyms

Synonyms Comment Organism
IPMDH
-
 Thermus thermophilus

Temperature Optimum [°C]

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

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
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]

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

pH Optimum

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

Cofactor

Cofactor Comment Organism Structure
NAD+
-
 Thermus thermophilus