Cloned (Comment) | Organism |
---|---|
idh2, DNA and amino acid sequence determination and anaysis, phylogenetic analysis, overexpression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain Rosetta (DE3) | Yarrowia lipolytica |
Protein Variants | Comment | Organism |
---|---|---|
R322D | site-directed mutagenesis, the mutant shows an 41fold higher Km for NADP+ than the wild-type enzyme, and it shows NAD+-dependent activity in contrast to the wild-type enzyme | Yarrowia lipolytica |
R322D/H323I | site-directed mutagenesis, inactive mutant | Yarrowia lipolytica |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetic analysis | Yarrowia lipolytica | |
0.031 | - |
NADP+ | pH 7.5, 25°C, recombinant enzyme, with Mg2+ | Yarrowia lipolytica | |
0.058 | - |
NADP+ | pH 7.5, 25°C, recombinant enzyme, with Mg2+ | Yarrowia lipolytica | |
0.059 | - |
isocitrate | pH 7.5, 25°C, recombinant enzyme, with Mg2+ | Yarrowia lipolytica | |
0.12 | - |
isocitrate | pH 7.5, 25°C, recombinant enzyme, with Mn2+ | Yarrowia lipolytica | |
47 | - |
NAD+ | pH 7.5, 25°C, recombinant enzyme mutant R322D | Yarrowia lipolytica |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | activates to 43% of the activity with Mg2+ | Yarrowia lipolytica | |
Li+ | activates to 24% of the activity with Mg2+ | Yarrowia lipolytica | |
Mg2+ | most favorable cofactor | Yarrowia lipolytica | |
Mn2+ | activates to 60% of the activity with Mg2+ | Yarrowia lipolytica | |
additional information | the enzyme is absolutely divalent cation dependent, no or poor activation by Ca2+, Zn2+, K+, Na+, and Rb+ | Yarrowia lipolytica | |
Ni2+ | activates to 10% of the activity with Mg2+ | Yarrowia lipolytica |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
45000 | - |
2 * 45000, recombinant enzyme, SDS-PAGE | Yarrowia lipolytica |
81300 | - |
recombinant enzyme, gel filtration | Yarrowia lipolytica |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
isocitrate + NADP+ | Yarrowia lipolytica | - |
2-oxoglutarate + CO2 + NADPH + H+ | - |
r | |
isocitrate + NADP+ | Yarrowia lipolytica CLIB 122 | - |
2-oxoglutarate + CO2 + NADPH + H+ | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Yarrowia lipolytica | Q6C2Y4 | - |
- |
Yarrowia lipolytica CLIB 122 | Q6C2Y4 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain Rosetta (DE3) b metal affinity chromatography | Yarrowia lipolytica |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
isocitrate + NAD+ | low activity with mutant enzyme R322D, no activity with the wild-type enzyme | Yarrowia lipolytica | 2-oxoglutarate + CO2 + NADH + H+ | - |
r | |
isocitrate + NAD+ | low activity with mutant enzyme R322D, no activity with the wild-type enzyme | Yarrowia lipolytica CLIB 122 | 2-oxoglutarate + CO2 + NADH + H+ | - |
r | |
isocitrate + NADP+ | - |
Yarrowia lipolytica | 2-oxoglutarate + CO2 + NADPH + H+ | - |
r | |
isocitrate + NADP+ | - |
Yarrowia lipolytica CLIB 122 | 2-oxoglutarate + CO2 + NADPH + H+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 45000, recombinant enzyme, SDS-PAGE | Yarrowia lipolytica |
Synonyms | Comment | Organism |
---|---|---|
NADP+-specific isocitrate dehydrogenase | - |
Yarrowia lipolytica |
YlIDP | - |
Yarrowia lipolytica |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
55 | - |
recombinant enzyme | Yarrowia lipolytica |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
45 | 55 | purified recombinant enzyme, stable up to 45°C, enzyme activity drops quickly above this temperature, incubation at 50°C for 20 min causes a 40% loss of activity, while incubation at 55°C almost inactivates the enzyme | Yarrowia lipolytica |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.38 | - |
NAD+ | pH 8.5, 55°C, recombinant mutant R322D | Yarrowia lipolytica | |
4.24 | - |
NADP+ | pH 7.5, 25°C, recombinant mutant R322D, with Mg2+ | Yarrowia lipolytica | |
72 | - |
NADP+ | pH 7.5, 25°C, recombinant wild-type enzyme, with Mg2+ | Yarrowia lipolytica |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | - |
recombinant enzyme | Yarrowia lipolytica |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | enzyme YlIDP is absolutely dependent on NADP+ and shows no NAD-dependent activity | Yarrowia lipolytica | |
NADP+ | - |
Yarrowia lipolytica | |
NADPH | - |
Yarrowia lipolytica |
General Information | Comment | Organism |
---|---|---|
evolution | phylogenetic analysis and evolutionary relationships between YlIDP and IDPs from other yeasts or yeast-related species | Yarrowia lipolytica |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.75 | - |
NADP+ | pH 7.5, 25°C, recombinant mutant R322D, with Mg2+ | Yarrowia lipolytica | |
1220 | - |
NADP+ | pH 7.5, 25°C, recombinant wild-type enzyme, with Mg2+ | Yarrowia lipolytica |