EC Number | Cloned (Comment) | Organism |
---|---|---|
1.1.1.42 | expressed in Escherichia coli Rosetta (DE3) cells | Methanococcoides methylutens |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
1.1.1.42 | H590L/R601D | the mutant with strongly decreased activity wit NADP+ compared to the wild type enzyme displays a 0.183fold preference for NAD+ over NADP+ | Methanococcoides methylutens |
1.1.1.42 | H590L/R601D/R650S | the mutant with severely decreased activity with NADP+ compared to the wild type enzyme displays a 5.93fold preference for NAD+ over NADP+ | Methanococcoides methylutens |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
1.1.1.42 | Ca2+ | in the presence of Mn2+, Ca2+ strongly inhibits the activity of the enzyme (28.52% residual activity) | Methanococcoides methylutens | |
1.1.1.42 | Zn2+ | in the presence of Mn2+, Zn2+ strongly inhibits the activity of the enzyme (42.59% residual activity) | Methanococcoides methylutens |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.1.1.42 | 0.0168 | - |
NADP+ | wild type enzyme, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens | |
1.1.1.42 | 0.4868 | - |
NADP+ | mutant enzyme H590L/R601D, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens | |
1.1.1.42 | 2.62 | - |
NAD+ | mutant enzyme H590L/R601D, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens | |
1.1.1.42 | 3.474 | - |
NAD+ | mutant enzyme H590L/R601D/R650S, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens | |
1.1.1.42 | 10.457 | - |
NADP+ | mutant enzyme H590L/R601D/R650S, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
1.1.1.42 | Mg2+ | Mg2+ can partially replace the activation of Mn2+ (40.3%). 2 mM used in assay conditions | Methanococcoides methylutens | |
1.1.1.42 | Mn2+ | Mn2+ is the most favored cation, 2 mM used in assay conditions | Methanococcoides methylutens | |
1.1.1.42 | additional information | Ni2+, Co2+, Na+, K+ and Li+ have almost no effect on the activity of the enzyme when assayed in the presence of Mn2+ | Methanococcoides methylutens |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.1.1.42 | isocitrate + NADP+ | Methanococcoides methylutens | - |
2-oxoglutarate + CO2 + NADPH + H+ | - |
? | |
1.1.1.42 | isocitrate + NADP+ | Methanococcoides methylutens MM1 | - |
2-oxoglutarate + CO2 + NADPH + H+ | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.1.1.42 | Methanococcoides methylutens | A0A0E3SSZ6 | - |
- |
1.1.1.42 | Methanococcoides methylutens MM1 | A0A0E3SSZ6 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
1.1.1.42 | Co2+ affinity resin column chromatography | Methanococcoides methylutens |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.1.1.42 | isocitrate + NAD+ | the reaction is catalyzed by mutant enzymes H590L/R601D/R650S and H590L/R601D | Methanococcoides methylutens | 2-oxoglutarate + CO2 + NADH + H+ | - |
? | |
1.1.1.42 | isocitrate + NAD+ | the reaction is catalyzed by mutant enzymes H590L/R601D/R650S and H590L/R601D | Methanococcoides methylutens MM1 | 2-oxoglutarate + CO2 + NADH + H+ | - |
? | |
1.1.1.42 | isocitrate + NADP+ | - |
Methanococcoides methylutens | 2-oxoglutarate + CO2 + NADPH + H+ | - |
? | |
1.1.1.42 | isocitrate + NADP+ | - |
Methanococcoides methylutens MM1 | 2-oxoglutarate + CO2 + NADPH + H+ | - |
? | |
1.1.1.42 | additional information | the enzyme is highly specific to NADP+ and cannot utilize NAD+ | Methanococcoides methylutens | ? | - |
- |
|
1.1.1.42 | additional information | the enzyme is highly specific to NADP+ and cannot utilize NAD+ | Methanococcoides methylutens MM1 | ? | - |
- |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
1.1.1.42 | monomer | 1 * 80000, SDS-PAGE | Methanococcoides methylutens |
1.1.1.42 | monomer | 1 * 81000, calculated from amino acid sequence | Methanococcoides methylutens |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.1.1.42 | IDH | - |
Methanococcoides methylutens |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.1.1.42 | 45 | - |
with Mn2+ | Methanococcoides methylutens |
1.1.1.42 | 50 | - |
with Mg2+ | Methanococcoides methylutens |
EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.1.1.42 | 35 | 39 | the enzyme is stable below 35°C. The enzyme sustains 50% activity after incubation at 39°C for 20 min with Mn2+ and 38°C with Mg2+ | Methanococcoides methylutens |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.1.1.42 | 52.3 | - |
NAD+ | mutant enzyme H590L/R601D, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens | |
1.1.1.42 | 53.1 | - |
NADP+ | mutant enzyme H590L/R601D, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens | |
1.1.1.42 | 55.5 | - |
NADP+ | wild type enzyme, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens | |
1.1.1.42 | 57.9 | - |
NADP+ | mutant enzyme H590L/R601D/R650S, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens | |
1.1.1.42 | 113.9 | - |
NAD+ | mutant enzyme H590L/R601D/R650S, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
1.1.1.42 | 8.2 | - |
with Mn2+ | Methanococcoides methylutens |
1.1.1.42 | 8.5 | - |
with Mg2+ | Methanococcoides methylutens |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
1.1.1.42 | NADP+ | the enzyme is highly specific for NADP+ | Methanococcoides methylutens |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.1.1.42 | 0.00553 | - |
NADP+ | mutant enzyme H590L/R601D/R650S, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens | |
1.1.1.42 | 0.01996 | - |
NAD+ | mutant enzyme H590L/R601D, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens | |
1.1.1.42 | 0.03278 | - |
NAD+ | mutant enzyme H590L/R601D/R650S, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens | |
1.1.1.42 | 0.109 | - |
NADP+ | mutant enzyme H590L/R601D, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens | |
1.1.1.42 | 3.303 | - |
NADP+ | wild type enzyme, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens |