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Literature summary extracted from

  • Suzuki, T.; Yasugi, M.; Arisaka, F.; Yamagishi, A.; Oshima, T.
    Adaptation of a thermophilic enzyme, 3-isopropylmalate dehydrogenase, to low temperatures (2001), Protein Eng., 14, 85-91.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.85 expression of wild-type and mutant enzymes in Escherichia coli Thermus thermophilus

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.85 A31G/G43A/A709G optimum temperature is shifted to lower temperatures compared to wild-type enzyme, activity is less temperature dependent than that of the wild-type, resulting in higher activity at temperatures below 60°C, improved turnover-numbers at 40°Cm increase in Km-values, slight loss of thermal stability Thermus thermophilus
1.1.1.85 C275T similar pH-dependence to that of the wild-type enzyme, improved binding affinities for both D-3-isopropylmalate and NAD+ result in an improved catalytic efficiency, mutant enzyme retains thermal stability Thermus thermophilus
1.1.1.85 G376A optimum temperature is shifted to lower temperatures compared to wild-type enzyme, activity is less temperature dependent than that of the wild-type, resulting in higher activity at temperatures below 60°C, improved turnover-numbers at 40°C, increase in Km-values, mutant enzyme retains thermal stability Thermus thermophilus
1.1.1.85 G43A optimum temperature is shifted to lower temperatures compared to wild-type enzyme, activity is less temperature dependent than that of the wild-type, resulting in higher activity at temperatures below 60°C, improved turnover-numbers at 40°C, increase ion Km-values, slight loss of thermal stability Thermus thermophilus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.85 0.001
-
 isopropylmalate pH 8.0, 40°C, mutant enzyme C275T Thermus thermophilus
1.1.1.85 0.0012
-
 isopropylmalate pH 8.0, 40°C, mutant enzyme G376A Thermus thermophilus
1.1.1.85 0.00125
-
 isopropylmalate pH 8.0, 40°C, wild-type enzyme Thermus thermophilus
1.1.1.85 0.0013
-
 isopropylmalate pH 8.0, 40°C, mutant enzyme A31G/G43A/A709G Thermus thermophilus
1.1.1.85 0.0024
-
 isopropylmalate pH 8.0, 40°C, mutant enzyme G43A Thermus thermophilus
1.1.1.85 0.0078
-
 isopropylmalate pH 8.0, 70°C, mutant enzyme C275T Thermus thermophilus
1.1.1.85 0.0089
-
 isopropylmalate pH 8.0, 70°C, wild-type enzyme Thermus thermophilus
1.1.1.85 0.0096
-
 isopropylmalate pH 8.0, 70°C, mutant enzyme A31G/G43A/A709G Thermus thermophilus
1.1.1.85 0.01
-
 isopropylmalate pH 8.0, 70°C, mutant enzyme G376A Thermus thermophilus
1.1.1.85 0.011
-
 isopropylmalate pH 8.0, 70°C, mutant enzyme G43A Thermus thermophilus
1.1.1.85 0.0149
-
 NAD+ pH 8.0, 40°C, reaction with 3-D-isopropylmalate, mutant enzyme C275T Thermus thermophilus
1.1.1.85 0.0258
-
 NAD+ pH 8.0, 40°C, isopropylmalate dehydrogenase wild-type Thermus thermophilus
1.1.1.85 0.157
-
 NAD+ pH 8.0, 40°C, mutant enzyme A31G/G43A/A709G Thermus thermophilus
1.1.1.85 0.16
-
 NAD+ pH 8.0, 40°C, mutant enzyme G43A Thermus thermophilus
1.1.1.85 0.225
-
 NAD+ pH 8.0, 40°C, isopropylmalate dehydrogenase cold-adapted mutant T1155 Thermus thermophilus
1.1.1.85 0.341
-
 NAD+ pH 8.0, 70°C, isopropylmalate wild-type Thermus thermophilus
1.1.1.85 0.82
-
 NAD+ at 70°C, mutant enzyme G43A Thermus thermophilus
1.1.1.85 0.917
-
 NAD+ at 70°C, mutant enzyme A31G/G43A/A709G Thermus thermophilus
1.1.1.85 1.01
-
 NAD+ at 40°C, isopropylmalate dehydrogenase cold-adapted mutant P215 Thermus thermophilus
1.1.1.85 3.56
-
 NAD+ 70°C, isopropylmalate dehydrogenase cold-adapted mutant P215 Thermus thermophilus

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.85 Thermus thermophilus
-
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.85 (2R,3S)-3-isopropylmalate + NAD+
-
 Thermus thermophilus 2-oxoisocaproate + NADH + H+ + CO2
-
 ?
1.1.1.85 (2R,3S)-3-isopropylmalate + NAD+
-
 Thermus thermophilus (2S)-2-isopropyl-3-oxosuccinate + NADH + H+
-
 r

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.85 82
-
 10 min, 50% inactivation, mutant enzyme A31G/G43A/A709G Thermus thermophilus
1.1.1.85 85
-
 10 min, 50% inactivation, mutant enzyme G43A Thermus thermophilus
1.1.1.85 87
-
 10 min, 50% inactivation, wild-type enzyme, mutant enzyme C275T and mutant enzyme G376A 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 2.36
-
 NAD+ pH 8.0, 40°C, wild-type enzyme Thermus thermophilus
1.1.1.85 2.36
-
 (2R,3S)-3-isopropylmalate pH 8.0, 40°C, wild-type enzyme Thermus thermophilus
1.1.1.85 2.41
-
 NAD+ pH 8.0, 40°C, mutant enzyme C275T Thermus thermophilus
1.1.1.85 2.41
-
 (2R,3S)-3-isopropylmalate pH 8.0, 40°C, mutant enzyme C275T Thermus thermophilus
1.1.1.85 7.2
-
 NAD+ pH 8.0, 40°C, mutant enzyme A31G/G43A/A709G Thermus thermophilus
1.1.1.85 7.2
-
 (2R,3S)-3-isopropylmalate pH 8.0, 40°C, mutant enzyme A31G/G43A/A709G Thermus thermophilus
1.1.1.85 10.7
-
 NAD+ pH 8.0, 40°C, mutant enzyme G43A Thermus thermophilus
1.1.1.85 10.7
-
 (2R,3S)-3-isopropylmalate pH 8.0, 40°C, mutant enzyme G43A Thermus thermophilus
1.1.1.85 17.7
-
 NAD+ pH 8.0, 40°C, mutant enzyme G376A Thermus thermophilus
1.1.1.85 17.7
-
 (2R,3S)-3-isopropylmalate pH 8.0, 40°C, mutant enzyme G376A Thermus thermophilus
1.1.1.85 44.7
-
 NAD+ pH 8.0, 70°C, mutant enzyme C275T Thermus thermophilus
1.1.1.85 44.7
-
 (2R,3S)-3-isopropylmalate pH 8.0, 70°C, mutant enzyme C275T Thermus thermophilus
1.1.1.85 45.8
-
 NAD+ pH 8.0, 70°C, wild-type enzyme Thermus thermophilus
1.1.1.85 45.8
-
 (2R,3S)-3-isopropylmalate pH 8.0, 70°C, wild-type enzyme Thermus thermophilus
1.1.1.85 50.1
-
 NAD+ pH 8.0, 70°C, mutant enzyme A31G/G43A/A709G Thermus thermophilus
1.1.1.85 50.1
-
 (2R,3S)-3-isopropylmalate pH 8.0, 70°C, mutant enzyme A31G/G43A/A709G Thermus thermophilus
1.1.1.85 63.6
-
 NAD+ pH 8.0, 70°C, mutant enzyme G43A Thermus thermophilus
1.1.1.85 63.6
-
 (2R,3S)-3-isopropylmalate pH 8.0, 70°C, mutant enzyme G43A Thermus thermophilus
1.1.1.85 100
-
 NAD+ pH 8.0, 70°C, mutant enzyme G376A Thermus thermophilus
1.1.1.85 100
-
 (2R,3S)-3-isopropylmalate pH 8.0, 70°C, mutant enzyme G376A Thermus thermophilus

Cofactor

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