Cloned (Comment) | Organism |
---|---|
- |
Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
D217E | less than 4% reductoisomerase activity in comparison to wild-type enzyme | Escherichia coli |
D217N | less than 4% reductoisomerase activity in comparison to wild-type enzyme | Escherichia coli |
E213D | 75% reductoisomerase activity in comparison of wild-type enzyme | Escherichia coli |
E213Q | less than 4% reductoisomerase activity in comparison of wild-type enzyme | Escherichia coli |
E221D | less than 4% reductoisomerase activity in comparison to wild-type enzyme | Escherichia coli |
E221Q | less than 4% reductoisomerase activity in comparison to wild-type enzyme | Escherichia coli |
E389D | less than 4% reductoisomerase activity in comparison to wild-type enzyme | Escherichia coli |
E389Q | less than 4% reductoisomerase activity in comparison to wild-type enzyme | Escherichia coli |
E393D | less than 4% reductoisomerase activity in comparison to wild-type enzyme | Escherichia coli |
E393Q | the mutant is insoluble, a soluble form is obtained only after denaturation | Escherichia coli |
H132K | less than 4% reductoisomerase activity in comparison of wild-type enzyme | Escherichia coli |
H132Q | less than 4% reductoisomerase activity in comparison of wild-type enzyme | Escherichia coli |
K155E | less than 4% reductoisomerase activity in comparison of wild-type enzyme | Escherichia coli |
K155Q | less than 4% reductoisomerase activity in comparison of wild-type enzyme | Escherichia coli |
K155R | less than 4% reductoisomerase activity in comparison of wild-type enzyme | Escherichia coli |
S414A | less than 4% reductoisomerase activity in comparison to wild-type enzyme | Escherichia coli |
S414T | less than 4% reductoisomerase activity in comparison to wild-type enzyme | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00016 | - |
NADPH | pH 8.0, 37°C, mutant E213D | Escherichia coli | |
0.00253 | - |
NADPH | pH 8.0, 37°C, wild-type enzyme | Escherichia coli | |
0.0031 | - |
NADPH | pH 8.0, 37°C, mutant H132K | Escherichia coli | |
0.0048 | - |
NADPH | pH 8.0, 37°C, mutant E393D | Escherichia coli | |
0.005 | - |
NADPH | pH 8.0, 37°C, mutant D217N | Escherichia coli | |
0.005 | - |
NADPH | pH 8.0, 37°C, mutant S414T | Escherichia coli | |
0.0073 | - |
NADPH | pH 8.0, 37°C, mutant K155R | Escherichia coli | |
0.00804 | - |
NADPH | pH 8.0, 37°C, mutant K155E | Escherichia coli | |
0.0084 | - |
NADPH | pH 8.0, 37°C, mutant S414A | Escherichia coli | |
0.0093 | - |
NADPH | pH 8.0, 37°C, mutant K155Q | Escherichia coli | |
0.02 | - |
NADPH | pH 8.0, 37°C, mutant E221D | Escherichia coli | |
0.02 | - |
NADPH | pH 8.0, 37°C, mutant E221Q | Escherichia coli | |
0.023 | - |
NADPH | pH 8.0, 37°C, mutant E389D | Escherichia coli | |
0.069 | - |
NADPH | pH 8.0, 37°C, mutant H132Q | Escherichia coli | |
0.08 | - |
NADPH | pH 8.0, 37°C, mutant D217E | Escherichia coli | |
0.17 | - |
2-ketopantoate | pH 8.0, 37°C, wild-type enzyme | Escherichia coli | |
0.21 | - |
3-hydroxy-2-ketobutyrate | pH 8.0, 37°C, wild-type enzyme | Escherichia coli | |
0.25 | - |
2-acetolactate | pH 8.0, 37°C, wild-type enzyme | Escherichia coli | |
0.27 | - |
3-hydroxy-3-methyl-2-ketobutyrate | pH 8.0, 37°C, wild-type enzyme | Escherichia coli | |
0.334 | - |
3-hydroxypyruvate | pH 8.0, 37°C, mutant S414A | Escherichia coli | |
0.356 | - |
2-acetolactate | pH 8.0, 37°C, mutant E221D | Escherichia coli | |
0.414 | - |
2-acetolactate | pH 8.0, 37°C, mutant S414T | Escherichia coli | |
0.441 | - |
3-hydroxypyruvate | pH 8.0, 37°C, mutant E213Q | Escherichia coli | |
0.588 | - |
3-hydroxypyruvate | pH 8.0, 37°C, mutant E393Q | Escherichia coli | |
0.711 | - |
2-acetolactate | pH 8.0, 37°C, mutant S414A | Escherichia coli | |
0.818 | - |
3-hydroxypyruvate | pH 8.0, 37°C, mutant H132K | Escherichia coli | |
0.922 | - |
2-acetolactate | pH 8.0, 37°C, mutant H213D | Escherichia coli | |
0.929 | - |
2-acetolactate | pH 8.0, 37°C, mutant H132Q | Escherichia coli | |
1.101 | - |
3-hydroxypyruvate | pH 8.0, 37°C, mutant S414T | Escherichia coli | |
1.218 | - |
2-acetolactate | pH 8.0, 37°C, mutant H155R | Escherichia coli | |
1.37 | - |
3-hydroxypyruvate | pH 8.0, 37°C, mutant E221D | Escherichia coli | |
1.54 | - |
pyruvate | pH 8.0, 37°C, wild-type enzyme | Escherichia coli | |
2.028 | - |
2-acetolactate | pH 8.0, 37°C, mutant E289D | Escherichia coli | |
2.66 | - |
3-hydroxypyruvate | pH 8.0, 37°C, mutant K155E | Escherichia coli | |
2.96 | - |
3-hydroxypyruvate | pH 8.0, 37°C, wild-type enzyme | Escherichia coli | |
3.15 | - |
2-ketovalerate | pH 8.0, 37°C, wild-type enzyme | Escherichia coli | |
3.32 | - |
3-hydroxypyruvate | pH 8.0, 37°C, mutant E393D | Escherichia coli | |
3.67 | - |
3-hydroxypyruvate | pH 8.0, 37°C, mutant E213D | Escherichia coli | |
4.56 | - |
2-Ketobutyrate | pH 8.0, 37°C, wild-type enzyme | Escherichia coli | |
6.91 | - |
2-ketoisovalerate | pH 8.0, 37°C, wild-type enzyme | Escherichia coli | |
7.43 | - |
3-hydroxypyruvate | pH 8.0, 37°C, mutant H132Q | Escherichia coli | |
7.64 | - |
3-hydroxypyruvate | pH 8.0, 37°C, mutant D217N | Escherichia coli | |
8.5 | - |
3-hydroxypyruvate | pH 8.0, 37°C, mutant E389D | Escherichia coli | |
8.88 | - |
3-hydroxypyruvate | pH 8.0, 37°C, mutant E389Q | Escherichia coli | |
13.6 | - |
3-hydroxypyruvate | pH 8.0, 37°C, mutant K155R | Escherichia coli | |
15.3 | - |
3-hydroxypyruvate | pH 8.0, 37°C, mutant K155Q | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | the reaction requires a divalent metal ion | Escherichia coli | |
Mg2+ | the reaction requires a divalent metal ion | Escherichia coli | |
Mn2+ | the reaction requires a divalent metal ion | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Escherichia coli | the enzyme is involved in the biosynthesis of the branched-chain amino acids | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Purification (Comment) | Organism |
---|---|
of wild-type and mutants | Escherichia coli |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
(2R)-2,3-dihydroxy-3-methylbutanoate + NADP+ = (2S)-2-hydroxy-2-methyl-3-oxobutanoate + NADPH + H+ | bifunctional enzyme that catalyses two different reactions at a common active site, an isomerization consisting of an alkyl migration, followed by an NADPH-dependent reduction of a-ketoacid | Escherichia coli |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
the specific activity with pyruvate is 1% and with 2-ketovalerate, 2-ketopantoate and 2-ketobutyrate is 8% of that of 2-acetolactate, comparison of activities of wild-type and mutant enzymes | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-acetolactate + NADPH + H+ | - |
Escherichia coli | 2,3-dihydroxy-3-methylbutanoate + NADP+ | - |
r | |
2-ketobutyrate + NADPH + H+ | - |
Escherichia coli | ? | - |
r | |
2-ketoisovalerate + NADPH + H+ | - |
Escherichia coli | ? | - |
r | |
2-ketopantoate + NADPH + H+ | - |
Escherichia coli | ? | - |
r | |
2-ketovalerate + NADPH + H+ | - |
Escherichia coli | ? | - |
r | |
3-hydroxy-2-ketobutyrate + NADPH + H+ | - |
Escherichia coli | ? | - |
r | |
3-hydroxy-3-methyl-2-ketobutyrate + NADP+ | - |
Escherichia coli | ? | - |
r | |
3-hydroxypyruvate + NADPH + H+ | - |
Escherichia coli | ? | - |
r | |
additional information | the enzyme is involved in the biosynthesis of the branched-chain amino acids | Escherichia coli | ? | - |
? | |
pyruvate + NADPH + H+ | - |
Escherichia coli | ? | - |
r |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.021 | - |
pyruvate | pH 8.0, 37°C | Escherichia coli | |
0.05 | - |
2-ketovalerate | pH 8.0, 37°C | Escherichia coli | |
0.167 | - |
2-Ketobutyrate | pH 8.0, 37°C | Escherichia coli | |
0.182 | - |
2-ketoisovalerate | pH 8.0, 37°C | Escherichia coli | |
0.194 | - |
2-ketopantoate | pH 8.0, 37°C | Escherichia coli | |
0.594 | - |
3-hydroxy-2-ketobutyrate | pH 8.0, 37°C | Escherichia coli | |
2.231 | - |
2-acetolactate | pH 8.0, 37°C | Escherichia coli | |
3.511 | - |
3-hydroxy-3-methyl-2-ketobutyrate | pH 8.0, 37°C | Escherichia coli | |
5.376 | - |
3-hydroxypyruvate | pH 8.0, 37°C | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | - |
Escherichia coli | |
NADPH | - |
Escherichia coli |