Inhibitors | Comment | Organism | Structure |
---|---|---|---|
K2SO4 | 250 mM K2SO4, pH 7.5, 20% inhibition | Cupriavidus necator | |
KCl | 400 mM KCl, pH 8.2, 20% inhibition | Cupriavidus necator |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mn2+ | 1 mM, 25% activation | Cupriavidus necator |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
100000 | - |
in presence of Leu, gel filtration | Cupriavidus necator |
114500 | - |
in absence of Leu, gel filtration | Cupriavidus necator |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-oxo-3-methylbutanoate + acetyl-CoA + H2O | Cupriavidus necator | first enzyme in biosynthesis of L-Leu | ? | - |
? | |
2-oxo-3-methylbutanoate + acetyl-CoA + H2O | Cupriavidus necator H 16 | first enzyme in biosynthesis of L-Leu | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Cupriavidus necator | - |
- |
- |
Cupriavidus necator | - |
Val-Ile double auxotrophic mutant of strain H 16 | - |
Cupriavidus necator H 16 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
- |
Cupriavidus necator |
Renatured (Comment) | Organism |
---|---|
irreversible denaturation after 5 min at 60°C | Cupriavidus necator |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
additional information | highest specific activity in cells harvested in the late exponential phase | Cupriavidus necator | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
12.05 | - |
- |
Cupriavidus necator |
Storage Stability | Organism |
---|---|
30°C, pH 7.5, stable for more than 10 days | Cupriavidus necator |
freezing at -20°C and thawing, about 10% loss of activity. Storage at -20°C, 100 mM potassium phosphate, 0.2 mM 2-oxo-3-methylbutanoate, pH 7.5 or pH 8.0, no further loss of activity after 5 months | Cupriavidus necator |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-oxo-3-methylbutanoate + acetyl-CoA + H2O | - |
Cupriavidus necator | 3-hydroxy-4-methyl-3-carboxypentanoate + CoA | - |
? | |
2-oxo-3-methylbutanoate + acetyl-CoA + H2O | - |
Cupriavidus necator H 16 | 3-hydroxy-4-methyl-3-carboxypentanoate + CoA | - |
? | |
2-oxo-3-methylbutanoate + acetyl-CoA + H2O | first enzyme in biosynthesis of L-Leu | Cupriavidus necator | ? | - |
? | |
2-oxo-3-methylbutanoate + acetyl-CoA + H2O | first enzyme in biosynthesis of L-Leu | Cupriavidus necator H 16 | ? | - |
? |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
44 | - |
- |
Cupriavidus necator |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
additional information | - |
cold lability can be overcome by stabilizing the enzyme by buffer of increased ionic strength | Cupriavidus necator |
additional information | - |
stabilization at low temperature with KCl | Cupriavidus necator |
60 | - |
5 min, in absence of substrates irreversible denaturation | Cupriavidus necator |
65 | - |
5 min, in presence of 2-oxo-3-methylbutanoate and acetyl-CoA, stable | Cupriavidus necator |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | 8.5 | Tris-HCl buffer | Cupriavidus necator |
8.2 | - |
phosphate buffer | Cupriavidus necator |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
7 | - |
3 h, about 15% of maximal activity | Cupriavidus necator |
7.2 | 8.2 | 3 h, stable | Cupriavidus necator |
9 | - |
3 h, about 50% loss of activity | Cupriavidus necator |
9.5 | - |
3 h, about 30% of maximal activity | Cupriavidus necator |