Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli | Thermus thermophilus |
Crystallization (Comment) | Organism |
---|---|
crystal structure of mutant H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M is determined at 2.4 A | Thermus thermophilus |
Protein Variants | Comment | Organism |
---|---|---|
E57V/R85M/Y86A/M208T/F217Y/V238M/R310M | no (2R,3S)-3-isopropylmalate dehydrogenase activity detected | Thermus thermophilus |
E57V/S72I/R85M/Y86A/F217Y/V238M/R310M | no (2R,3S)-3-isopropylmalate dehydrogenase activity detected | Thermus thermophilus |
E57V/S72I/R85M/Y86A/M208T/F217Y/R310M | in comparison to mutant H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M this mutant shows a significant decrease in kcat ((2R,3S)-3-isopropylmalate), alteration of Km value only moderate | Thermus thermophilus |
E57V/S72I/R85M/Y86A/M208T/F217Y/V238M | in comparison to mutant H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M this mutant shows a significant decrease in kcat ((2R,3S)-3-isopropylmalate), alteration of Km value only moderate | Thermus thermophilus |
E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M | in comparison to mutant H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M this mutant shows a larger kcat ((2R,3S)-3-isopropylmalate) and a larger Km((2R,3S)-3-isopropylmalate) or (NAD+) temperatures giving half of the initial activity is 87.8 °C | Thermus thermophilus |
E57V/S72I/R85M/Y86A/M208T/V238M/R310M | in comparison to mutant H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M this mutant shows a significant decrease in kcat ((2R,3S)-3-isopropylmalate), alteration of Km value only moderate | Thermus thermophilus |
E57V/S72I/R85V/Y86A/M208T/F217Y/V238M/R310M | no (2R,3S)-3-isopropylmalate dehydrogenase activity detected | Thermus thermophilus |
H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M | TtHICDH variant with considerably higher IPMDH activity than double mutant R85V/Y86A, Km ((2R,3S)-3-isopropylmalate) or (NAD+) lower than wild-type HICDH but still considerably higher than wild-type (2R,3S)-3-isopropylmalate dehydrogenase, kcat ((2R,3S)-3-isopropylmalate) much higher than wild-type HICDH but still considerably lower than wild-type (2R,3S)-3-isopropylmalate dehydrogenase, temperatures giving half of the initial activity is 79.9 °C, crystal structure determined at 2.4 A, mutant is a homotetramer | Thermus thermophilus |
R85V/Y86A | TtHICDH variant with high IPMDH activity, Km ((2R,3S)-3-isopropylmalate) or (NAD+) lower than wild-type HICDH but still considerably higher than wild-type (2R,3S)-3-isopropylmalate dehydrogenase, kcat ((2R,3S)-3-isopropylmalate) higher than wild-type HICDH but still considerably lower than wild-type (2R,3S)-3-isopropylmalate dehydrogenase, temperatures giving half of the initial activity is 92.5 °C | Thermus thermophilus |
S72I/R85M/Y86A/M208T/F217Y/V238M/R310M | in comparison to mutant H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M this mutant shows a significant decrease in kcat ((2R,3S)-3-isopropylmalate), alteration of Km value only moderate | Thermus thermophilus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.59 | - |
(2R,3S)-3-isopropylmalate | mutant H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C | Thermus thermophilus | |
0.9 | - |
(2R,3S)-3-isopropylmalate | mutant E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C | Thermus thermophilus | |
0.9 | - |
NAD+ | mutant E57V/S72I/R85M/Y86A/M208T/V238M/R310M, pH 8.0, 37°C | Thermus thermophilus | |
0.9 | - |
(2R,3S)-3-isopropylmalate | mutant E57V/S72I/R85M/Y86A/M208T/V238M/R310M, pH 8.0, 37°C | Thermus thermophilus | |
1 | - |
(2R,3S)-3-isopropylmalate | mutant S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C | Thermus thermophilus | |
1 | - |
(2R,3S)-3-isopropylmalate | wild-type (2R,3S)-3-isopropylmalate dehydrogenase, pH 8.0, 37°C | Thermus thermophilus | |
1 | - |
(2R,3S)-3-isopropylmalate | wild-type HICDH, pH 8.0, 37°C | Thermus thermophilus | |
1.2 | - |
NAD+ | mutant E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C | Thermus thermophilus | |
1.2 | - |
NAD+ | mutant H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C | Thermus thermophilus | |
1.5 | - |
(2R,3S)-3-isopropylmalate | mutant E57V/S72I/R85M/Y86A/M208T/F217Y/V238M, pH 8.0, 37°C | Thermus thermophilus | |
1.6 | - |
NAD+ | mutant E57V/S72I/R85M/Y86A/M208T/F217Y/V238M, pH 8.0, 37°C | Thermus thermophilus | |
1.6 | - |
NAD+ | mutant S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C | Thermus thermophilus | |
1.7 | - |
NAD+ | mutant E57V/S72I/R85M/Y86A/M208T/F217Y/R310M, pH 8.0, 37°C | Thermus thermophilus | |
1.7 | - |
NAD+ | mutant R85V/Y86A, pH 8.0, 37°C | Thermus thermophilus | |
1.9 | - |
NAD+ | wild-type (2R,3S)-3-isopropylmalate dehydrogenase, pH 8.0, 37°C | Thermus thermophilus | |
1.9 | - |
NAD+ | wild-type HICDH, pH 8.0, 37°C | Thermus thermophilus | |
1.9 | - |
(2R,3S)-3-isopropylmalate | mutant E57V/S72I/R85M/Y86A/M208T/F217Y/R310M, pH 8.0, 37°C | Thermus thermophilus | |
2.6 | - |
(2R,3S)-3-isopropylmalate | mutant R85V/Y86A, pH 8.0, 37°C | Thermus thermophilus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Thermus thermophilus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermus thermophilus | Q72IW9 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(2R,3S)-3-isopropylmalate + NAD+ | mutant enzymes with increased activity | Thermus thermophilus | (2S)-2-isopropyl-3-oxosuccinate + NADH + H+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
IPMDH | - |
Thermus thermophilus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Thermus thermophilus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.08 | - |
(2R,3S)-3-isopropylmalate | wild-type HICDH, pH 8.0, 37°C | Thermus thermophilus | |
0.3 | - |
(2R,3S)-3-isopropylmalate | mutant E57V/S72I/R85M/Y86A/M208T/V238M/R310M, pH 8.0, 37°C | Thermus thermophilus | |
0.4 | - |
(2R,3S)-3-isopropylmalate | mutant R85V/Y86A, pH 8.0, 37°C | Thermus thermophilus | |
0.6 | - |
(2R,3S)-3-isopropylmalate | mutant E57V/S72I/R85M/Y86A/M208T/F217Y/V238M, pH 8.0, 37°C | Thermus thermophilus | |
1.6 | - |
(2R,3S)-3-isopropylmalate | mutant S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C | Thermus thermophilus | |
1.8 | - |
(2R,3S)-3-isopropylmalate | mutant E57V/S72I/R85M/Y86A/M208T/F217Y/R310M, pH 8.0, 37°C | Thermus thermophilus | |
2.9 | - |
(2R,3S)-3-isopropylmalate | mutant H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C | Thermus thermophilus | |
5.1 | - |
(2R,3S)-3-isopropylmalate | mutant E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C | Thermus thermophilus | |
5.7 | - |
(2R,3S)-3-isopropylmalate | wild-type (2R,3S)-3-isopropylmalate dehydrogenase, pH 8.0, 37°C | Thermus thermophilus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Thermus thermophilus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | - |
Thermus thermophilus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.12 | - |
NAD+ | wild-type (2R,3S)-3-isopropylmalate dehydrogenase, pH 8.0, 37°C | Thermus thermophilus | |
0.16 | - |
(2R,3S)-3-isopropylmalate | mutant R85V/Y86A, pH 8.0, 37°C | Thermus thermophilus | |
0.18 | - |
(2R,3S)-3-isopropylmalate | wild-type (2R,3S)-3-isopropylmalate dehydrogenase, pH 8.0, 37°C | Thermus thermophilus | |
0.23 | - |
NAD+ | mutant R85V/Y86A, pH 8.0, 37°C | Thermus thermophilus | |
0.25 | - |
NAD+ | mutant H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C | Thermus thermophilus | |
0.33 | - |
NAD+ | mutant E57V/S72I/R85M/Y86A/M208T/V238M/R310M, pH 8.0, 37°C | Thermus thermophilus | |
0.33 | - |
(2R,3S)-3-isopropylmalate | mutant E57V/S72I/R85M/Y86A/M208T/V238M/R310M, pH 8.0, 37°C | Thermus thermophilus | |
0.38 | - |
NAD+ | mutant E57V/S72I/R85M/Y86A/M208T/F217Y/V238M, pH 8.0, 37°C | Thermus thermophilus | |
0.4 | - |
(2R,3S)-3-isopropylmalate | mutant E57V/S72I/R85M/Y86A/M208T/F217Y/V238M, pH 8.0, 37°C | Thermus thermophilus | |
0.41 | - |
NAD+ | wild-type HICDH, pH 8.0, 37°C | Thermus thermophilus | |
0.49 | - |
(2R,3S)-3-isopropylmalate | mutant H15Y/E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C | Thermus thermophilus | |
0.75 | - |
(2R,3S)-3-isopropylmalate | wild-type HICDH, pH 8.0, 37°C | Thermus thermophilus | |
0.94 | - |
(2R,3S)-3-isopropylmalate | mutant E57V/S72I/R85M/Y86A/M208T/F217Y/R310M, pH 8.0, 37°C | Thermus thermophilus | |
1.1 | - |
NAD+ | mutant E57V/S72I/R85M/Y86A/M208T/F217Y/R310M , pH 8.0, 37°C | Thermus thermophilus | |
1.4 | - |
NAD+ | mutant S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C | Thermus thermophilus | |
1.6 | - |
(2R,3S)-3-isopropylmalate | mutant S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C | Thermus thermophilus | |
4.2 | - |
NAD+ | mutant E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C | Thermus thermophilus | |
5.4 | - |
(2R,3S)-3-isopropylmalate | mutant E57V/S72I/R85M/Y86A/M208T/F217Y/V238M/R310M, pH 8.0, 37°C | Thermus thermophilus |