Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli Rosetta (DE3) cells | Methanococcoides methylutens |
Protein Variants | Comment | Organism |
---|---|---|
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 |
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 |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | in the presence of Mn2+, Ca2+ strongly inhibits the activity of the enzyme (28.52% residual activity) | Methanococcoides methylutens | |
Zn2+ | in the presence of Mn2+, Zn2+ strongly inhibits the activity of the enzyme (42.59% residual activity) | Methanococcoides methylutens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0168 | - |
NADP+ | wild type enzyme, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens | |
0.4868 | - |
NADP+ | mutant enzyme H590L/R601D, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens | |
2.62 | - |
NAD+ | mutant enzyme H590L/R601D, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens | |
3.474 | - |
NAD+ | mutant enzyme H590L/R601D/R650S, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens | |
10.457 | - |
NADP+ | mutant enzyme H590L/R601D/R650S, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | Mg2+ can partially replace the activation of Mn2+ (40.3%). 2 mM used in assay conditions | Methanococcoides methylutens | |
Mn2+ | Mn2+ is the most favored cation, 2 mM used in assay conditions | Methanococcoides methylutens | |
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 |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
isocitrate + NADP+ | Methanococcoides methylutens | - |
2-oxoglutarate + CO2 + NADPH + H+ | - |
? | |
isocitrate + NADP+ | Methanococcoides methylutens MM1 | - |
2-oxoglutarate + CO2 + NADPH + H+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Methanococcoides methylutens | A0A0E3SSZ6 | - |
- |
Methanococcoides methylutens MM1 | A0A0E3SSZ6 | - |
- |
Purification (Comment) | Organism |
---|---|
Co2+ affinity resin column chromatography | Methanococcoides methylutens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
isocitrate + NAD+ | the reaction is catalyzed by mutant enzymes H590L/R601D/R650S and H590L/R601D | Methanococcoides methylutens | 2-oxoglutarate + CO2 + NADH + H+ | - |
? | |
isocitrate + NAD+ | the reaction is catalyzed by mutant enzymes H590L/R601D/R650S and H590L/R601D | Methanococcoides methylutens MM1 | 2-oxoglutarate + CO2 + NADH + H+ | - |
? | |
isocitrate + NADP+ | - |
Methanococcoides methylutens | 2-oxoglutarate + CO2 + NADPH + H+ | - |
? | |
isocitrate + NADP+ | - |
Methanococcoides methylutens MM1 | 2-oxoglutarate + CO2 + NADPH + H+ | - |
? | |
additional information | the enzyme is highly specific to NADP+ and cannot utilize NAD+ | Methanococcoides methylutens | ? | - |
- |
|
additional information | the enzyme is highly specific to NADP+ and cannot utilize NAD+ | Methanococcoides methylutens MM1 | ? | - |
- |
Subunits | Comment | Organism |
---|---|---|
monomer | 1 * 80000, SDS-PAGE | Methanococcoides methylutens |
monomer | 1 * 81000, calculated from amino acid sequence | Methanococcoides methylutens |
Synonyms | Comment | Organism |
---|---|---|
IDH | - |
Methanococcoides methylutens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
45 | - |
with Mn2+ | Methanococcoides methylutens |
50 | - |
with Mg2+ | Methanococcoides methylutens |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
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 |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
52.3 | - |
NAD+ | mutant enzyme H590L/R601D, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens | |
53.1 | - |
NADP+ | mutant enzyme H590L/R601D, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens | |
55.5 | - |
NADP+ | wild type enzyme, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens | |
57.9 | - |
NADP+ | mutant enzyme H590L/R601D/R650S, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens | |
113.9 | - |
NAD+ | mutant enzyme H590L/R601D/R650S, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.2 | - |
with Mn2+ | Methanococcoides methylutens |
8.5 | - |
with Mg2+ | Methanococcoides methylutens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | the enzyme is highly specific for NADP+ | Methanococcoides methylutens |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00553 | - |
NADP+ | mutant enzyme H590L/R601D/R650S, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens | |
0.01996 | - |
NAD+ | mutant enzyme H590L/R601D, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens | |
0.03278 | - |
NAD+ | mutant enzyme H590L/R601D/R650S, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens | |
0.109 | - |
NADP+ | mutant enzyme H590L/R601D, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens | |
3.303 | - |
NADP+ | wild type enzyme, in the presence of Mn2+, at pH 8.2 and 45°C | Methanococcoides methylutens |