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Literature summary for 1.1.1.42 extracted from
- Analysis of amino acid residues involved in the thermal properties of isocitrate dehydrogenases from a psychrophilic bacterium, Colwellia maris, and a psychrotrophic bacterium, Pseudomonas psychrophila (2020), J. Biosci. Bioeng., 129, 284-290 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expressed in enzyme-defective Escherichia coli strain DEK2004 | Colwellia maris |
| expressed in enzyme-defective Escherichia coli strain DEK2004 | Pseudomonas psychrophila |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| A741S | the mutant exhibits higher specific activity and thermostability than the wild type enzyme | Colwellia maris |
| E538L | the mutant exhibits higher thermostability than wild type enzyme | Colwellia maris |
| E596L | the mutant exhibits higher specific activity and thermostability than the wild type enzyme | Colwellia maris |
| L536E | the mutant is more thermolabile than wild type enzyme | Pseudomonas psychrophila |
| L594E | the mutant is more thermolabile than wild type enzyme and has a lower specific activity at temperatures over 45°C | Pseudomonas psychrophila |
| S739A | the mutant is more thermolabile than wild type enzyme | Pseudomonas psychrophila |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.01558 | - |
isocitrate | mutant enzyme L594E, at pH 8.0 and 20°C | Pseudomonas psychrophila |  |
| 0.02008 | - |
isocitrate | wild type enzyme, at pH 8.0 and 20°C | Pseudomonas psychrophila | |
| 0.04666 | - |
isocitrate | mutant enzyme E596L, at pH 8.0 and 20°C | Colwellia maris | |
| 0.06 | - |
isocitrate | wild type enzyme, at pH 8.0 and 20°C | Colwellia maris | |
| 0.06742 | - |
isocitrate | mutant enzyme A741S, at pH 8.0 and 20°C | Colwellia maris | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mg2+ | 2 mM used in assay conditions | Colwellia maris | |
| Mg2+ | 2 mM used in assay conditions | Pseudomonas psychrophila | |
| Mn2+ | 0.67 mM used in assay conditions | Colwellia maris | |
| Mn2+ | 0.67 mM used in assay conditions | Pseudomonas psychrophila | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| isocitrate + NADP+ | Colwellia maris | - |
2-oxoglutarate + CO2 + NADPH + H+ | - |
? | |
| isocitrate + NADP+ | Pseudomonas psychrophila | - |
2-oxoglutarate + CO2 + NADPH + H+ | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Colwellia maris | - |
- |
- |
| Pseudomonas psychrophila | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| Ni-NTA agarose column chromatography | Colwellia maris |
| Ni-NTA agarose column chromatography | Pseudomonas psychrophila |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| isocitrate + NADP+ | - |
Colwellia maris | 2-oxoglutarate + CO2 + NADPH + H+ | - |
? | |
| isocitrate + NADP+ | - |
Pseudomonas psychrophila | 2-oxoglutarate + CO2 + NADPH + H+ | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| monomer | 1 * 80000, His-tagged enzyme, SDS-PAGE | Colwellia maris |
| monomer | 1 * 80000, His-tagged enzyme, SDS-PAGE | Pseudomonas psychrophila |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| IDH | - |
Colwellia maris |
| IDH | - |
Pseudomonas psychrophila |
| NADP+-dependent isocitrate dehydrogenase | - |
Colwellia maris |
| NADP+-dependent isocitrate dehydrogenase | - |
Pseudomonas psychrophila |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | - |
- |
Colwellia maris |
| 50 | - |
- |
Pseudomonas psychrophila |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | 50 | after incubation at 25°C and 30°C, the enzyme activity loses 31% and 69% of ist activity, respectively, and no activity is detected after incubation at 50°C | Colwellia maris |
| 40 | 70 | after incubation at temperatures up to 40°C, the wild type enzyme maintains its activity, but its residual activity is decreased by incubation at over 45°C and is lost after incubation at 70°C | Pseudomonas psychrophila |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 69.88 | - |
isocitrate | wild type enzyme, at pH 8.0 and 20°C | Colwellia maris | |
| 82.47 | - |
isocitrate | mutant enzyme A741S, at pH 8.0 and 20°C | Colwellia maris | |
| 89.04 | - |
isocitrate | mutant enzyme E596L, at pH 8.0 and 20°C | Colwellia maris | |
| 136.85 | - |
isocitrate | wild type enzyme, at pH 8.0 and 20°C | Pseudomonas psychrophila | |
| 158.91 | - |
isocitrate | mutant enzyme L594E, at pH 8.0 and 20°C | Pseudomonas psychrophila | |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| NADP+ | - |
Colwellia maris | |
| NADP+ | - |
Pseudomonas psychrophila | |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 1164 | - |
isocitrate | wild type enzyme, at pH 8.0 and 20°C | Colwellia maris | |
| 1227 | - |
isocitrate | mutant enzyme A741S, at pH 8.0 and 20°C | Colwellia maris | |
| 1914 | - |
isocitrate | mutant enzyme E596L, at pH 8.0 and 20°C | Colwellia maris | |
| 6810 | - |
isocitrate | wild type enzyme, at pH 8.0 and 20°C | Pseudomonas psychrophila | |
| 101970 | - |
isocitrate | mutant enzyme L594E, at pH 8.0 and 20°C | Pseudomonas psychrophila | |
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