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Literature summary for 1.1.1.85 extracted from
- Enhancement of latent 3-isopropylmalate dehydrogenase activity of promiscuous homoisocitrate dehydrogenase by directed evolution (2010), Biochem. J., 431, 401-410.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expressed in Escherichia coli | Thermus thermophilus |
Crystallization (Commentary)
| 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
| 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 [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
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mg2+ | - |
Thermus thermophilus | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Thermus thermophilus | Q72IW9 | - |
- |
Substrates and Products (Substrate)
| 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
| Synonyms | Comment | Organism |
|---|---|---|
| IPMDH | - |
Thermus thermophilus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
assay at | Thermus thermophilus |
Turnover Number [1/s]
| 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
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8 | - |
assay at | Thermus thermophilus |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| NADH | - |
Thermus thermophilus | |
kcat/KM [mM/s]
| 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 | |
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