Cloned (Comment) | Organism |
---|---|
gene KT71_12905, DNA and amino acid sequence determination and analysis, sequence comparisons,, phylogenetic analysis and tree, recombinant expression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain Rosetta (DE3) | Congregibacter litoralis |
Protein Variants | Comment | Organism |
---|---|---|
D487R/L488H | site-directed mutagenesis, the coenzyme specificity of a the ClIDH mutant is altered compared to the wild-type enzyme, and the preference of the mutant for NADP+ is approximately 24fold higher than that for NAD+, suggesting that ClIDH is an NAD+-specific ancestral enzyme in the type II IDH subgroup | Congregibacter litoralis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.157 | - |
NADP+ | mutant D487R/L488H, with Mn2+, pH 7.5, 35°C | Congregibacter litoralis | |
0.263 | - |
NAD+ | wild-type enzyme, with Mg2+, pH 7.5, 35°C | Congregibacter litoralis | |
0.309 | - |
NAD+ | wild-type enzyme, with Mn2+, pH 7.5, 35°C | Congregibacter litoralis | |
0.664 | - |
NAD+ | mutant D487R/L488H, with Mn2+, pH 7.5, 35°C | Congregibacter litoralis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | activates, 38.3% activity compared to Mn2+ | Congregibacter litoralis | |
Mg2+ | activates, 45.7% activity compared to Mn2+ | Congregibacter litoralis | |
Mn2+ | most effectivly activating cation | Congregibacter litoralis | |
additional information | enzyme ClIDH is dependent on divalent cations, the most effective being Mn2+. Poor activation by Ca2+, Zn2+, Ni2+, K+, Na+, Rb+, and Li+. No activation by Cu2+ | Congregibacter litoralis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
64000 | - |
- |
Congregibacter litoralis |
390000 | - |
recombinant His6-tagged enzyme, gel filtration and analytical ultracentrifugation | Congregibacter litoralis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
isocitrate + NAD+ | Congregibacter litoralis | - |
2-oxoglutarate + CO2 + NADH + H+ | - |
? | |
isocitrate + NAD+ | Congregibacter litoralis KT71 | - |
2-oxoglutarate + CO2 + NADH + H+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Congregibacter litoralis | A4ADB8 | - |
- |
Congregibacter litoralis KT71 | A4ADB8 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain Rosetta (DE3) by metal affinity chromatography | Congregibacter litoralis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
isocitrate + NAD+ | - |
Congregibacter litoralis | 2-oxoglutarate + CO2 + NADH + H+ | - |
? | |
isocitrate + NAD+ | - |
Congregibacter litoralis KT71 | 2-oxoglutarate + CO2 + NADH + H+ | - |
? | |
isocitrate + NADP+ | activity with enzyme mutant D487R/L488H, not the wild-type enzyme | Congregibacter litoralis | 2-oxoglutarate + CO2 + NADPH + H+ | - |
? | |
isocitrate + NADP+ | activity with enzyme mutant D487R/L488H, not the wild-type enzyme | Congregibacter litoralis KT71 | 2-oxoglutarate + CO2 + NADPH + H+ | - |
? |
Subunits | Comment | Organism |
---|---|---|
homohexamer | 6 * 64000, recombinant His6-tagged enzyme, SDS-PAGE | Congregibacter litoralis |
Synonyms | Comment | Organism |
---|---|---|
type II NAD+-specific isocitrate dehydrogenase | - |
Congregibacter litoralis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
35 | - |
- |
Congregibacter litoralis |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
20 | 32 | activity range, profile overview | Congregibacter litoralis |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
33 | - |
purified recombinant enzyme, 20 min, loss of 50% activity | Congregibacter litoralis |
38 | - |
purified recombinant enzyme, 20 min, loss of over 90% activity | Congregibacter litoralis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
9.7 | - |
NAD+ | mutant D487R/L488H, with Mn2+, pH 7.5, 35°C | Congregibacter litoralis | |
36.7 | - |
NAD+ | wild-type enzyme, with Mg2+, pH 7.5, 35°C | Congregibacter litoralis | |
55.1 | - |
NADP+ | mutant D487R/L488H, with Mn2+, pH 7.5, 35°C | Congregibacter litoralis | |
84.7 | - |
NAD+ | wild-type enzyme, with Mn2+, pH 7.5, 35°C | Congregibacter litoralis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
- |
Congregibacter litoralis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | the recombinant enzyme is NAD+-specific and shows no detectable activity with NADP+ | Congregibacter litoralis |
General Information | Comment | Organism |
---|---|---|
evolution | phylogenetic analyses divide the IDH protein family into two subgroups: types I and II. Based on cofactor usage, IDHs are either NAD+-specific (NAD-IDH) or NADP+-specific (NADP-IDH). NADP-IDH evolved from NAD-IDH. Type I IDHs include NAD-IDHs and NADP-IDHs. Type II NAD-IDHs is identified from the marine bacterium Congregibacter litoralis KT71, i.e ClIDH. Evolutionary relationships between 151 IDHs from different organisms, overview | Congregibacter litoralis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
15 | - |
NAD+ | mutant D487R/L488H, with Mn2+, pH 7.5, 35°C | Congregibacter litoralis | |
140 | - |
NAD+ | wild-type enzyme, with Mg2+, pH 7.5, 35°C | Congregibacter litoralis | |
270 | - |
NAD+ | wild-type enzyme, with Mn2+, pH 7.5, 35°C | Congregibacter litoralis | |
350 | - |
NADP+ | mutant D487R/L488H, with Mn2+, pH 7.5, 35°C | Congregibacter litoralis |