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 | Comment | Organism | Structure |
---|---|---|---|
NADPH | - |
Escherichia coli |
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 | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Escherichia coli |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Purification (Comment) | Organism |
---|---|
wild-type and mutant enzyme | Escherichia coli |
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 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 | Comment | Organism | Structure |
---|---|---|---|
NADPH | - |
Escherichia coli |
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 |