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Literature summary for 1.1.1.383 extracted from
- General approach to reversing ketol-acid reductoisomerase cofactor dependence from NADPH to NADH (2013), Proc. Natl. Acad. Sci. USA, 110, 10946-10951.
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| A71S/R76D/S78AD/Q110V | mutant engineered for reversal in cofactor specificity for NADH over NADPH | Shewanella sp. |
| A71S/R76D/S78D/Q110V | mutant engineered for reversal in cofactor specificity for NADH over NADPH | Escherichia coli |
| A71S/R76D/S78D/Q110V/D146G/G185R/K433E | mutant engineered for reversal in cofactor specificity for NADH over NADPH | Escherichia coli |
| additional information | recipe for altering the cofactor specificity of the ketol-acid reductoisomerases (KARIs). Combining results for an engineered NADH-dependent variant of Escherichia coli KARI with available KARI crystal structures and a comprehensive KARI-sequence alignment, key cofactor specificity determinants are identified and used to construct KARIs with reversed cofactor preference | Escherichia coli |
| R76D/Ser78D | mutant engineered for reversal in cofactor specificity for NADH over NADPH | Shewanella sp. |
| S61D/S63D | mutant engineered for reversal in cofactor specificity for NADH over NADPH | Slackia exigua |
| S61D/S63D/I95V | mutant engineered for reversal in cofactor specificity for NADH over NADPH | Slackia exigua |
| V48L/R49P/K52E/S53D | mutant engineered for reversal in cofactor specificity for NADH over NADPH | Lactococcus lactis subsp. lactis |
| V48L/R49P/K52L/S53D | mutant engineered for reversal in cofactor specificity for NADH over NADPH | Lactococcus lactis subsp. lactis |
| V48L/R49P/K52L/S53D/E59K/T182S/E320K | mutant engineered for reversal in cofactor specificity for NADH over NADPH | Lactococcus lactis subsp. lactis |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.001 | - |
NADPH | wild-type, pH not specified in the publication, temperature not specified in the publication | Shewanella sp. |  |
| 0.001 | - |
NADPH | wild-type, pH not specified in the publication, temperature not specified in the publication | Slackia exigua | |
| 0.013 | - |
NADPH | wild-type, pH not specified in the publication, temperature not specified in the publication | Lactococcus lactis subsp. lactis | |
| 0.015 | - |
NADH | mutant V48L/R49P/K52L/S53D/E59K/T182S/E320K, pH not specified in the publication, temperature not specified in the publication | Lactococcus lactis subsp. lactis | |
| 0.026 | - |
NADH | mutant A71S/R76D/S78D/Q110V/D146G/G185R/K433E , pH not specified in the publication, temperature not specified in the publication | Escherichia coli | |
| 0.03 | - |
NADH | mutant A71S/R76D/S78D/Q110V, pH not specified in the publication, temperature not specified in the publication | Escherichia coli | |
| 0.041 | - |
NADPH | wild-type, pH not specified in the publication, temperature not specified in the publication | Escherichia coli | |
| 0.045 | - |
NADH | wild-type, pH not specified in the publication, temperature not specified in the publication | Slackia exigua | |
| 0.047 | - |
NADH | mutant S61D/S63D/I95V, pH not specified in the publication, temperature not specified in the publication | Slackia exigua | |
| 0.075 | - |
NADH | mutant A71S/R76D/S78AD/Q110V, pH not specified in the publication, temperature not specified in the publication | Shewanella sp. | |
| 0.09 | - |
NADH | mutant R76D/Ser78D, pH not specified in the publication, temperature not specified in the publication | Shewanella sp. | |
| 0.108 | - |
NADH | mutant V48L/R49P/K52L/S53D, pH not specified in the publication, temperature not specified in the publication | Lactococcus lactis subsp. lactis | |
| 0.113 | - |
NADH | mutant S61D/S63D, pH not specified in the publication, temperature not specified in the publication | Slackia exigua | |
| 0.128 | - |
NADH | mutant V48L/R49P/K52E/S53D, pH not specified in the publication, temperature not specified in the publication | Lactococcus lactis subsp. lactis | |
| 0.285 | - |
NADH | wild-type, pH not specified in the publication, temperature not specified in the publication | Lactococcus lactis subsp. lactis | |
| 0.415 | - |
NADH | wild-type, pH not specified in the publication, temperature not specified in the publication | Shewanella sp. | |
| 0.6 | - |
NADPH | mutant A71S/R76D/S78AD/Q110V, pH not specified in the publication, temperature not specified in the publication | Shewanella sp. | |
| 0.65 | - |
NADPH | mutant A71S/R76D/S78D/Q110V, pH not specified in the publication, temperature not specified in the publication | Escherichia coli | |
| 0.75 | - |
NADPH | mutant V48L/R49P/K52L/S53D/E59K/T182S/E320K, pH not specified in the publication, temperature not specified in the publication | Lactococcus lactis subsp. lactis | |
| 0.88 | - |
NADPH | mutant S61D/S63D, pH not specified in the publication, temperature not specified in the publication | Slackia exigua | |
| 1 | - |
NADPH | mutant R76D/Ser78D, pH not specified in the publication, temperature not specified in the publication | Shewanella sp. | |
| 1 | - |
NADPH | mutant S61D/S63D/I95V, pH not specified in the publication, temperature not specified in the publication | Slackia exigua | |
| 1 | - |
NADPH | mutant V48L/R49P/K52L/S53D, pH not specified in the publication, temperature not specified in the publication | Lactococcus lactis subsp. lactis | |
| 1.075 | - |
NADH | wild-type, pH not specified in the publication, temperature not specified in the publication | Escherichia coli | |
| 1.18 | - |
NADPH | mutant V48L/R49P/K52E/S53D, pH not specified in the publication, temperature not specified in the publication | Lactococcus lactis subsp. lactis | |
| 1.4 | - |
NADPH | mutant A71S/R76D/S78D/Q110V/D146G/G185R/K433E , pH not specified in the publication, temperature not specified in the publication | Escherichia coli | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Escherichia coli | P05793 | - |
- |
| Lactococcus lactis subsp. lactis | Q02138 | - |
- |
| Lactococcus lactis subsp. lactis IL1403 | Q02138 | - |
- |
| Shewanella sp. | A0KS29 | - |
- |
| Shewanella sp. ANA-3 | A0KS29 | - |
- |
| Slackia exigua | D0WGK0 | - |
- |
| Slackia exigua DSM 15923 | D0WGK0 | - |
- |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| (S)-2-acetolactate + NADH + H+ | - |
Escherichia coli | 2,3-dihydroxy-3-methylbutanoate + NAD+ | - |
? | |
| (S)-2-acetolactate + NADH + H+ | - |
Shewanella sp. | 2,3-dihydroxy-3-methylbutanoate + NAD+ | - |
? | |
| (S)-2-acetolactate + NADH + H+ | - |
Slackia exigua | 2,3-dihydroxy-3-methylbutanoate + NAD+ | - |
? | |
| (S)-2-acetolactate + NADH + H+ | - |
Lactococcus lactis subsp. lactis | 2,3-dihydroxy-3-methylbutanoate + NAD+ | - |
? | |
| (S)-2-acetolactate + NADH + H+ | - |
Slackia exigua DSM 15923 | 2,3-dihydroxy-3-methylbutanoate + NAD+ | - |
? | |
| (S)-2-acetolactate + NADH + H+ | - |
Shewanella sp. ANA-3 | 2,3-dihydroxy-3-methylbutanoate + NAD+ | - |
? | |
| (S)-2-acetolactate + NADH + H+ | - |
Lactococcus lactis subsp. lactis IL1403 | 2,3-dihydroxy-3-methylbutanoate + NAD+ | - |
? | |
| (S)-2-acetolactate + NADPH + H+ | - |
Escherichia coli | 2,3-dihydroxy-3-methylbutanoate + NADP+ | - |
? | |
| (S)-2-acetolactate + NADPH + H+ | - |
Shewanella sp. | 2,3-dihydroxy-3-methylbutanoate + NADP+ | - |
? | |
| (S)-2-acetolactate + NADPH + H+ | - |
Slackia exigua | 2,3-dihydroxy-3-methylbutanoate + NADP+ | - |
? | |
| (S)-2-acetolactate + NADPH + H+ | - |
Lactococcus lactis subsp. lactis | 2,3-dihydroxy-3-methylbutanoate + NADP+ | - |
? | |
| (S)-2-acetolactate + NADPH + H+ | - |
Slackia exigua DSM 15923 | 2,3-dihydroxy-3-methylbutanoate + NADP+ | - |
? | |
| (S)-2-acetolactate + NADPH + H+ | - |
Shewanella sp. ANA-3 | 2,3-dihydroxy-3-methylbutanoate + NADP+ | - |
? | |
| (S)-2-acetolactate + NADPH + H+ | - |
Lactococcus lactis subsp. lactis IL1403 | 2,3-dihydroxy-3-methylbutanoate + NADP+ | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| ilvC | - |
Slackia exigua |
| ilvC | - |
Lactococcus lactis subsp. lactis |
| Shewana3_0355 | - |
Shewanella sp. |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.06 | - |
NADPH | mutant V48L/R49P/K52E/S53D, pH not specified in the publication, temperature not specified in the publication | Lactococcus lactis subsp. lactis | |
| 0.08 | - |
NADPH | mutant V48L/R49P/K52L/S53D, pH not specified in the publication, temperature not specified in the publication | Lactococcus lactis subsp. lactis | |
| 0.1 | - |
NADH | wild-type, pH not specified in the publication, temperature not specified in the publication | Lactococcus lactis subsp. lactis | |
| 0.1 | - |
NADPH | mutant R76D/Ser78D, pH not specified in the publication, temperature not specified in the publication | Shewanella sp. | |
| 0.1 | - |
NADPH | mutant S61D/S63D, pH not specified in the publication, temperature not specified in the publication | Slackia exigua | |
| 0.2 | - |
NADPH | mutant A71S/R76D/S78D/Q110V, pH not specified in the publication, temperature not specified in the publication | Escherichia coli | |
| 0.25 | - |
NADPH | mutant S61D/S63D/I95V, pH not specified in the publication, temperature not specified in the publication | Slackia exigua | |
| 0.3 | - |
NADH | wild-type, pH not specified in the publication, temperature not specified in the publication | Escherichia coli | |
| 0.3 | - |
NADPH | mutant A71S/R76D/S78AD/Q110V, pH not specified in the publication, temperature not specified in the publication | Shewanella sp. | |
| 0.35 | - |
NADH | mutant V48L/R49P/K52E/S53D, pH not specified in the publication, temperature not specified in the publication | Lactococcus lactis subsp. lactis | |
| 0.35 | - |
NADPH | mutant V48L/R49P/K52L/S53D/E59K/T182S/E320K, pH not specified in the publication, temperature not specified in the publication | Lactococcus lactis subsp. lactis | |
| 0.4 | - |
NADH | mutant V48L/R49P/K52L/S53D, pH not specified in the publication, temperature not specified in the publication | Lactococcus lactis subsp. lactis | |
| 0.41 | - |
NADH | wild-type, pH not specified in the publication, temperature not specified in the publication | Slackia exigua | |
| 0.54 | - |
NADPH | mutant A71S/R76D/S78D/Q110V/D146G/G185R/K433E , pH not specified in the publication, temperature not specified in the publication | Escherichia coli | |
| 0.8 | - |
NADPH | wild-type, pH not specified in the publication, temperature not specified in the publication | Slackia exigua | |
| 0.8 | - |
NADPH | wild-type, pH not specified in the publication, temperature not specified in the publication | Lactococcus lactis subsp. lactis | |
| 0.97 | - |
NADH | mutant S61D/S63D, pH not specified in the publication, temperature not specified in the publication | Slackia exigua | |
| 1.01 | - |
NADH | mutant S61D/S63D/I95V, pH not specified in the publication, temperature not specified in the publication | Slackia exigua | |
| 1.07 | - |
NADH | mutant V48L/R49P/K52L/S53D/E59K/T182S/E320K, pH not specified in the publication, temperature not specified in the publication | Lactococcus lactis subsp. lactis | |
| 1.1 | - |
NADH | wild-type, pH not specified in the publication, temperature not specified in the publication | Shewanella sp. | |
| 1.3 | - |
NADH | mutant R76D/Ser78D, pH not specified in the publication, temperature not specified in the publication | Shewanella sp. | |
| 2.3 | - |
NADH | mutant A71S/R76D/S78D/Q110V, pH not specified in the publication, temperature not specified in the publication | Escherichia coli | |
| 2.4 | - |
NADH | mutant A71S/R76D/S78AD/Q110V, pH not specified in the publication, temperature not specified in the publication | Shewanella sp. | |
| 3.6 | - |
NADPH | wild-type, pH not specified in the publication, temperature not specified in the publication | Escherichia coli | |
| 4.3 | - |
NADH | mutant A71S/R76D/S78D/Q110V/D146G/G185R/K433E , pH not specified in the publication, temperature not specified in the publication | Escherichia coli | |
| 4.5 | - |
NADPH | wild-type, pH not specified in the publication, temperature not specified in the publication | Shewanella sp. | |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.05 | - |
NADPH | mutant V48L/R49P/K52E/S53D, pH not specified in the publication, temperature not specified in the publication | Lactococcus lactis subsp. lactis | |
| 0.08 | - |
NADPH | mutant V48L/R49P/K52L/S53D, pH not specified in the publication, temperature not specified in the publication | Lactococcus lactis subsp. lactis | |
| 0.11 | - |
NADPH | mutant S61D/S63D, pH not specified in the publication, temperature not specified in the publication | Slackia exigua | |
| 0.17 | - |
NADPH | mutant R76D/Ser78D, pH not specified in the publication, temperature not specified in the publication | Shewanella sp. | |
| 0.25 | - |
NADPH | mutant S61D/S63D/I95V, pH not specified in the publication, temperature not specified in the publication | Slackia exigua | |
| 0.3 | - |
NADH | wild-type, pH not specified in the publication, temperature not specified in the publication | Escherichia coli | |
| 0.4 | - |
NADPH | mutant A71S/R76D/S78D/Q110V, pH not specified in the publication, temperature not specified in the publication | Escherichia coli | |
| 0.4 | - |
NADPH | mutant A71S/R76D/S78D/Q110V/D146G/G185R/K433E , pH not specified in the publication, temperature not specified in the publication | Escherichia coli | |
| 0.43 | - |
NADH | wild-type, pH not specified in the publication, temperature not specified in the publication | Lactococcus lactis subsp. lactis | |
| 0.47 | - |
NADPH | mutant V48L/R49P/K52L/S53D/E59K/T182S/E320K, pH not specified in the publication, temperature not specified in the publication | Lactococcus lactis subsp. lactis | |
| 0.5 | - |
NADPH | mutant A71S/R76D/S78AD/Q110V, pH not specified in the publication, temperature not specified in the publication | Shewanella sp. | |
| 2.6 | - |
NADH | wild-type, pH not specified in the publication, temperature not specified in the publication | Shewanella sp. | |
| 2.7 | - |
NADH | mutant V48L/R49P/K52E/S53D, pH not specified in the publication, temperature not specified in the publication | Lactococcus lactis subsp. lactis | |
| 3.7 | - |
NADH | mutant V48L/R49P/K52L/S53D, pH not specified in the publication, temperature not specified in the publication | Lactococcus lactis subsp. lactis | |
| 9 | - |
NADH | wild-type, pH not specified in the publication, temperature not specified in the publication | Slackia exigua | |
| 9 | - |
NADH | mutant S61D/S63D, pH not specified in the publication, temperature not specified in the publication | Slackia exigua | |
| 14 | - |
NADH | mutant R76D/Ser78D, pH not specified in the publication, temperature not specified in the publication | Shewanella sp. | |
| 22 | - |
NADH | mutant S61D/S63D/I95V, pH not specified in the publication, temperature not specified in the publication | Slackia exigua | |
| 32 | - |
NADH | mutant A71S/R76D/S78AD/Q110V, pH not specified in the publication, temperature not specified in the publication | Shewanella sp. | |
| 65 | - |
NADPH | wild-type, pH not specified in the publication, temperature not specified in the publication | Lactococcus lactis subsp. lactis | |
| 71 | - |
NADH | mutant V48L/R49P/K52L/S53D/E59K/T182S/E320K, pH not specified in the publication, temperature not specified in the publication | Lactococcus lactis subsp. lactis | |
| 74 | - |
NADH | mutant A71S/R76D/S78D/Q110V, pH not specified in the publication, temperature not specified in the publication | Escherichia coli | |
| 88 | - |
NADPH | wild-type, pH not specified in the publication, temperature not specified in the publication | Escherichia coli | |
| 165 | - |
NADH | mutant A71S/R76D/S78D/Q110V/D146G/G185R/K433E , pH not specified in the publication, temperature not specified in the publication | Escherichia coli | |
| 800 | - |
NADPH | wild-type, pH not specified in the publication, temperature not specified in the publication | Slackia exigua | |
| 4500 | - |
NADPH | wild-type, pH not specified in the publication, temperature not specified in the publication | Shewanella sp. | |
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