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Literature summary for 1.1.1.87 extracted from
- Identification of a novel trifunctional homoisocitrate dehydrogenase and modulation of the broad substrate specificity through site-directed mutagenesis (2005), Biochem. Biophys. Res. Commun., 336, 596-602.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| DNA and amino acid sequence determination and analysis, overexpression in Escherichia coli strain OM17 | Deinococcus radiodurans |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| A80del | site-directed mutagenesis, the mutant shows altered substrate specificity preferring isocitrate to homoisocitrate, it is unable to oxidize 3-isopropylmalate, the specificity is similar to the enzyme from Thermus thermophilus | Deinococcus radiodurans |
| additional information | modulation of the broad substrate specificity of the trifunctional enzyme through site-directed mutagenesis, overview | Deinococcus radiodurans |
| R87T | site-directed mutagenesis, the mutant oxidizes homoisocitrate, but not isocitrate and 3-isopropylmalate | Deinococcus radiodurans |
| R87V | site-directed mutagenesis, the mutant oxidizes homoisocitrate, but not isocitrate and 3-isopropylmalate | Deinococcus radiodurans |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.211 | - |
homoisocitrate | pH 7.8, 25°C, recombinant wild-type enzyme | Deinococcus radiodurans |  |
| 0.291 | - |
isocitrate | pH 7.8, 25°C, recombinant wild-type enzyme | Deinococcus radiodurans | |
| 0.465 | - |
isocitrate | pH 7.8, 25°C, recombinant mutant A80del | Deinococcus radiodurans | |
| 0.819 | - |
homoisocitrate | pH 7.8, 25°C, recombinant mutant A80del | Deinococcus radiodurans | |
| 1 | - |
homoisocitrate | pH 7.8, 25°C, recombinant mutant R87V | Deinococcus radiodurans | |
| 1.33 | - |
3-isopropylmalate | pH 7.8, 25°C, recombinant wild-type enzyme | Deinococcus radiodurans | |
| 1.5 | - |
homoisocitrate | pH 7.8, 25°C, recombinant mutant R87T | Deinococcus radiodurans | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| K+ | required | Deinococcus radiodurans | |
| Mn2+ | required | Deinococcus radiodurans | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 40000 | - |
4 * 40000, recombinant enzyme, SDS-PAGE | Deinococcus radiodurans |
| 154000 | - |
recombinant enzyme, gel filtration | Deinococcus radiodurans |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Deinococcus radiodurans | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant enzyme from Escherichia coli strain OM17 by anion exchange and hydrophobic interaction chromatography, followed by another step of anion exchange chromatography | Deinococcus radiodurans |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| additional information | - |
- |
Deinococcus radiodurans |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 3-isopropylmalate + NAD+ | - |
Deinococcus radiodurans | ? | - |
? | |
| homoisocitrate + NAD+ | 1.5fold preferred to isocitrate | Deinococcus radiodurans | ? | - |
? | |
| isocitrate + NAD+ | - |
Deinococcus radiodurans | ? | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| tetramer | 4 * 40000, recombinant enzyme, SDS-PAGE | Deinococcus radiodurans |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| HICDH | - |
Deinococcus radiodurans |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 25 | - |
assay at | Deinococcus radiodurans |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.371 | - |
3-isopropylmalate | pH 7.8, 25°C, recombinant wild-type enzyme | Deinococcus radiodurans | |
| 5.46 | - |
homoisocitrate | pH 7.8, 25°C, recombinant mutant A80del | Deinococcus radiodurans | |
| 10.9 | - |
isocitrate | pH 7.8, 25°C, recombinant mutant A80del | Deinococcus radiodurans | |
| 20.8 | - |
homoisocitrate | pH 7.8, 25°C, recombinant mutant R87V | Deinococcus radiodurans | |
| 27.8 | - |
homoisocitrate | pH 7.8, 25°C, recombinant mutant R87T | Deinococcus radiodurans | |
| 42.8 | - |
isocitrate | pH 7.8, 25°C, recombinant wild-type enzyme | Deinococcus radiodurans | |
| 46.2 | - |
homoisocitrate | pH 7.8, 25°C, recombinant wild-type enzyme | Deinococcus radiodurans | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.8 | - |
assay at | Deinococcus radiodurans |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| NAD+ | - |
Deinococcus radiodurans | |
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