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Literature summary for 1.1.1.86 extracted from

  • Rane, M.J.; Calvo, K.C.
    Reversal of the nucleotide specificity of ketol acid reductoisomerase by site-directed mutagenesis identifies the NADPH binding site (1997), Arch. Biochem. Biophys., 338, 83-89.
    View publication on PubMed

Protein Variants

Protein Variants Comment Organism
K69L Km-value for NADP+ in the reaction with 2,3-dihydroxy-3-methylbutanoate is 2.1fold higher than the Km-value of the wild-type enzyme, the turnover-number for the reaction with NADP+ and 2,3-dihydroxy-3-methylbutanoate is lower by a factor 163 compared to wild-type enzyme. Km-value for NADPH in the reaction with acetolactate is comparable to that of the wild-type emzyme, the turnover-number for the reaction with NADP+ and 2,3-dihydroxy-3-methylbutanoate is 1.8fold higher than that of the wild-type enzyme Escherichia coli
K75Q Km-value for NADP+ in the reaction with 2,3-dihydroxy-3-methylbutanoate is 2.7fold higher than the Km-value of the wild-type enzyme, the turnover-number for the reaction with NADP+ and 2,3-dihydroxy-3-methylbutanoate is lower by a factor 77.5 compared to wild-type enzyme. Km-value for NADPH in the reaction with acetolactate is lower by a factor 2.9 compared to the Km-value of the wild-type enzyme, the turnover-number for the reaction with NADP+ and 2,3-dihydroxy-3-methylbutanoate is lower by a factor 12.9 compared to wild-type enzyme Escherichia coli
R68D/K69L/K75V/R76D turnover-number for reaction with NADH and acetolactate is 48fold higher compared to wild-type enzyme, turnover-number for reaction with NADPH and acetolactate is lower by factor 3.7 compared to wild-type enzyme, Km-value for NADH in the reaction with NADH and acetolactate is lower by a factor 10.8 compared to wild-type enzyme, Km-value for NADH in the reaction with NADPH and acetolactate is 30fold higher compared to wild-type enzyme Escherichia coli
R68Q Km-value for NADP+ in the reaction with 2,3-dihydroxy-3-methylbutanoate is 6.9fold higher than the Km-value of the wild-type enzyme, the turnover-number for the reaction with NADP+ and 2,3-dihydroxy-3-methylbutanoate is lower by a factor 345 compared to wild-type enzyme. Km-value for NADPH in the reaction with acetolactate is 3.4fold higher thahn that of the wild-type enzyme, the turnover-number for the reaction with NADP+ and 2,3-dihydroxy-3-methylbutanoate is lower by a factor 18 compared to wild-type enzyme Escherichia coli
R76D turnover-number for reaction with NADH and acetolactate is 48fold higher compared to wild-type enzyme, turnover-number for reaction with NADPH and acetolactate is lower by factor 4 compared to wild-type enzyme, Km-value for NADH in the reaction with NADH and acetolactate is lower by a factor 2.5 compared to wild-type enzyme, Km-value for NADPH in the reaction with NADPH and acetolactate is 55fold higher compared to wild-type enzyme Escherichia coli
R76Q Km-value for NADP+ in the reaction with 2,3-dihydroxy-3-methylbutanoate is 17.1fold higher than the Km-value of the wild-type enzyme, the turnover-number for the reaction with NADP+ and 2,3-dihydroxy-3-methylbutanoate is lower by a factor 258 compared to wild-type enzyme. Km-value for NADPH in the reaction with acetolactate is 5fold higher than that of the wild-type enzyme, the turnover-number for the reaction with NADP+ and 2,3-dihydroxy-3-methylbutanoate is lower by a factor 19.5 compared to wild-type enzyme Escherichia coli
R76Q/R68A turnover-number for reaction with NADH and acetolactate is 20fold higher compared to wild-type enzyme, turnover-number for reaction with NADPH and acetolactate is lower by factor 28 compared to wild-type enzyme, Km-value for NADH in the reaction with NADH and acetolactate is comparable to that of wild-type enzyme, Km-value for NADPH in the reaction with NADPH and acetolactate is 22fold higher compared to wild-type enzyme Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
NADPH
-
Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0025
-
NADPH pH 8.0, 22°C, reaction with acetolactate, mutant enzyme K75Q Escherichia coli
0.007
-
NADPH pH 8.0, 22°C, wild type enzyme Escherichia coli
0.0076
-
NADPH pH 8.0, 22°C, reaction with acetolactate, mutant enzyme K69L Escherichia coli
0.0089
-
NADP+ pH 8.0, 22°C, reaction with 2,3-dihydroxy-3-methylbutanoate, mutant enzyme K69L Escherichia coli
0.0112
-
NADP+ pH 8.0, 22°C, reaction with 2,3-dihydroxy-3-methylbutanoate, mutant enzyme K75Q Escherichia coli
0.019
-
NADH pH 8.0, 22°C, mutant enzyme R68D/K69L/K75V/R76D Escherichia coli
0.019
-
NADH mutant enzyme R68D/K69L/K75V/R76D Escherichia coli
0.0245
-
NADPH pH 8.0, 22°C, reaction with 2-acetolactate, mutant enzyme R68QL Escherichia coli
0.029
-
NADP+ pH 8.0, 22°C, reaction with 2,3-dihydroxy-3-methylbutanoate, mutant enzyme R68Q Escherichia coli
0.0365
-
NADPH pH 8.0, 22°C, reaction with 2-acetolactate, mutant enzyme R76Q Escherichia coli
0.072
-
NADP+ pH 8.0, 22°C, reaction with 2,3-dihydroxy-3-methylbutanoate, mutant enzyme R76Q Escherichia coli
0.082
-
NADH pH 8, 22°C, mutant enzyme R76D Escherichia coli
0.082
-
NADH mutant enzyme R76D Escherichia coli
0.16
-
NADPH pH 8, 22°C, mutant enzyme R76Q/R68A Escherichia coli
0.193
-
NADH pH 8, 22°C, mutant enzyme R76Q/R68A Escherichia coli
0.193
-
NADH mutant enzyme R76Q/R68A Escherichia coli
0.207
-
NADH wild type enzyme Escherichia coli
0.207
-
NADH pH 8.0, 22°C, wild type enzyme Escherichia coli
0.222
-
NADPH pH 8, 22°C, mutant enzyme R68D/K69L/K75V/R76D Escherichia coli
0.401
-
NADPH pH 8, 22°C, mutant enzyme R76D Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2-acetolactate + NADPH Escherichia coli enzyme of branched chain amino acid synthesis 2,3-dihydroxy-3-methylbutanoate + NADP+
-
r

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-

Purification (Commentary)

Purification (Comment) Organism
wild-type and mutant enzyme Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-acetolactate + NADH + H+
-
Escherichia coli 2,3-dihydroxy-3-methylbutanoate + NAD+ + H+
-
r
2-acetolactate + NADPH enzyme of branched chain amino acid synthesis Escherichia coli 2,3-dihydroxy-3-methylbutanoate + NADP+
-
r
NADPH + 2-acetolactate
-
Escherichia coli NADP+ + 3-hydroxy-3-methyl-2-oxobutyrate
-
?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0000933
-
NADPH pH 8.0, 22°C, reaction with acetolactate, mutant enzyme K75Q Escherichia coli
0.00015
-
NADP+ pH 8.0, 22°C, reaction with 2,3-dihydroxy-3-methylbutanoate, mutant enzyme R68Q Escherichia coli
0.0002
-
NADP+ pH 8.0, 22°C, reaction with 2,3-dihydroxy-3-methylbutanoate, mutant enzyme R76Q Escherichia coli
0.000317
-
NADP+ pH 8.0, 22°C, reaction with 2,3-dihydroxy-3-methylbutanoate, mutant enzyme K69L Escherichia coli
0.000667
-
NADP+ pH 8.0, 22°C, reaction with 2,3-dihydroxy-3-methylbutanoate, mutant enzyme K75Q Escherichia coli
0.00183
-
NADH wild type enzyme Escherichia coli
0.00433
-
NADPH pH 8, 22°C, mutant enzyme R76Q/R68A Escherichia coli
0.00617
-
NADPH pH 8.0, 22°C, reaction with acetolactate, mutant enzyme R76Q Escherichia coli
0.00667
-
NADPH pH 8.0, 22°C, reaction with acetolactate, mutant enzyme R68Q Escherichia coli
0.03
-
NADPH pH 8, 22°C, mutant enzyme R76D Escherichia coli
0.0328
-
NADPH pH 8.0, 22°C, mutant enzyme R68D/K69L/K75V/R76D Escherichia coli
0.0367
-
NADH pH 8, 22°C, mutant enzyme R76Q/R68A Escherichia coli
0.0883
-
NADH pH 8, 22°C, mutant enzyme R68D/K69L/K75V/R76D and mutant enzyme R76D Escherichia coli
0.12
-
NADPH pH 8, 22°C, wild type enzyme Escherichia coli
0.212
-
NADPH pH 8.0, 22°C, reaction with acetolactate, mutant enzyme K69L Escherichia coli

Cofactor

Cofactor Comment Organism Structure
NADPH
-
Escherichia coli

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.0012
-
NADPH pH 8.0, 22°C, wild-type enzyme Escherichia coli
0.0018
-
NADPH pH 8.0, 22°C, mutant enzyme K69L Escherichia coli
0.0039
-
NADPH pH 8.0, 22°C, mutant enzyme K75Q Escherichia coli
0.0085
-
NADPH pH 8.0, 22°C, mutant enzyme R68Q Escherichia coli
0.0226
-
NADPH pH 8.0, 22°C, mutant enzyme K76Q Escherichia coli