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 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 | 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 | 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 | Comment | Organism |
---|---|---|
ilvC | - |
Slackia exigua |
ilvC | - |
Lactococcus lactis subsp. lactis |
Shewana3_0355 | - |
Shewanella sp. |
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 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. |