Cloned (Comment) | Organism |
---|---|
- |
Pseudomonas straminea |
Protein Variants | Comment | Organism |
---|---|---|
C124S | kcat/Km for 4-oxalomesaconate is 1.9fold higher than wild-type value, mutant enzyme is less stable than wild-type enzyme at 4°C in 20 mM potassium phosphate buffer containing 1 mM DTT, mutant enzyme is less stable than wild-type enzyme at 4°C in 20 mM potassium phosphate buffer containing 1 mM DTT | Pseudomonas straminea |
C131S | kcat/Km for 4-oxalomesaconate is 26% higher than wild-type value, mutant enzyme is less stable than wild-type enzyme at 4°C in 20 mM potassium phosphate buffer containing 1 mM DTT | Pseudomonas straminea |
C183S | kcat/Km for 4-oxalomesaconate is 1.8fold higher than wild-type value, mutant enzyme is less stable than wild-type enzyme at 4°C in 20 mM potassium phosphate buffer containing 1 mM DTT | Pseudomonas straminea |
C186S | kcat/Km for 4-oxalomesaconate is 16% higher than wild-type value, mutant enzyme is more stable than wild-type enzyme at 4°C in 20 mM potassium phosphate buffer containing 1 mM DTT | Pseudomonas straminea |
C205S | kcat/Km for 4-oxalomesaconate is 29% higher than wild-type value, mutant enzyme is less stable than wild-type enzyme at 4°C in 20 mM potassium phosphate buffer containing 1 mM DTT | Pseudomonas straminea |
C260S | kcat/Km for 4-oxalomesaconate is 18% higher than wild-type value, mutant enzyme is less stable than wild-type enzyme at 4°C in 20 mM potassium phosphate buffer containing 1 mM DTT | Pseudomonas straminea |
C92S | kcat/Km for 4-oxalomesaconate is 18% higher than wild-type value, mutant enzyme is less stable than wild-type enzyme at 4°C in 20 mM potassium phosphate buffer containing 1 mM DTT | Pseudomonas straminea |
C96S | kcat/Km for 4-oxalomesaconate is 3.9fold lower than wild-type value, mutant enzyme is less stable than wild-type enzyme at 4°C in 20 mM potassium phosphate buffer containing 1 mM DTT | Pseudomonas straminea |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
5,5'-dithiobis(2-nitrobenzoic acid) | inhibition is primarily due to modification of Cys186. DTNB-treated C186S remains fully active | Pseudomonas straminea | |
HgCl2 | 1 mM, all the mutant enzymes lose activity with incubation (C92S, C96S, C124S, C131S, C183S, C186S, C205S, C260S) | Pseudomonas straminea | |
Zn2+ | - |
Pseudomonas straminea |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0029 | - |
4-oxalomesaconate | 25°C, pH 8.0, mutant enzyme C92S | Pseudomonas straminea | |
0.00618 | - |
4-oxalomesaconate | 25°C, pH 8.0, mutant enzyme C205S | Pseudomonas straminea | |
0.00733 | - |
4-oxalomesaconate | 25°C, pH 8.0, mutant enzyme C260S | Pseudomonas straminea | |
0.00794 | - |
4-oxalomesaconate | 25°C, pH 8.0, mutant enzyme C124S | Pseudomonas straminea | |
0.00838 | - |
4-oxalomesaconate | 25°C, pH 8.0, mutant enzyme C131S | Pseudomonas straminea | |
0.00851 | - |
4-oxalomesaconate | 25°C, pH 8.0, mutant enzyme C183S | Pseudomonas straminea | |
0.00994 | - |
4-oxalomesaconate | 25°C, pH 8.0, mutant enzyme C186S | Pseudomonas straminea | |
0.0107 | - |
4-oxalomesaconate | 25°C, pH 8.0, wild-type enzyme | Pseudomonas straminea | |
0.087 | - |
4-oxalomesaconate | 25°C, pH 8.0, mutant enzyme C96S | Pseudomonas straminea |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Zn2+ | PocOMH possesses one Zn2+ per subunit. It strongly binds to the enzyme protein | Pseudomonas straminea |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
38000 | - |
2 * 38000, SDS-PAGE | Pseudomonas straminea |
72000 | - |
gel filtration | Pseudomonas straminea |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas straminea | - |
- |
- |
Pseudomonas straminea NGJ1 | - |
- |
- |
Oxidation Stability | Organism |
---|---|
the enzyme is unstable in absence of reducing reagents | Pseudomonas straminea |
Purification (Comment) | Organism |
---|---|
recombinant | Pseudomonas straminea |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
113 | - |
- |
Pseudomonas straminea |
Storage Stability | Organism |
---|---|
-25°C, 20 mM potassium phosphate buffer containing 1 mM DTT, stable for at least 2 months | Pseudomonas straminea |
4°C, 20 mM potassium phosphate buffer containing 1 mM DTT, wild-type enzyme loses 20% of initial activity after 1 d | Pseudomonas straminea |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-oxalomesaconate | - |
Pseudomonas straminea | ? | - |
? | |
4-oxalomesaconate | - |
Pseudomonas straminea NGJ1 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 38000, SDS-PAGE | Pseudomonas straminea |
Synonyms | Comment | Organism |
---|---|---|
4-oxalomesaconate hydratase | - |
Pseudomonas straminea |
PocOMH | - |
Pseudomonas straminea |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.2 | - |
4-oxalomesaconate | 25°C, pH 8.0, mutant enzyme C131S | Pseudomonas straminea | |
2.3 | - |
4-oxalomesaconate | 25°C, pH 8.0, wild-type enzyme | Pseudomonas straminea | |
24.1 | - |
4-oxalomesaconate | 25°C, pH 8.0, mutant enzyme C92S | Pseudomonas straminea | |
33.9 | - |
4-oxalomesaconate | 25°C, pH 8.0, mutant enzyme C183S | Pseudomonas straminea | |
56 | - |
4-oxalomesaconate | 25°C, pH 8.0, mutant enzyme C205S | Pseudomonas straminea | |
60.8 | - |
4-oxalomesaconate | 25°C, pH 8.0, mutant enzyme C260S | Pseudomonas straminea | |
74.1 | - |
4-oxalomesaconate | 25°C, pH 8.0, mutant enzyme C131S | Pseudomonas straminea | |
75.4 | - |
4-oxalomesaconate | 25°C, pH 8.0, wild-type enzyme | Pseudomonas straminea | |
81.3 | - |
4-oxalomesaconate | 25°C, pH 8.0, mutant enzyme C186S | Pseudomonas straminea | |
104 | - |
4-oxalomesaconate | 25°C, pH 8.0, mutant enzyme C124S | Pseudomonas straminea | |
159 | - |
4-oxalomesaconate | 25°C, pH 8.0, mutant enzyme C96S | Pseudomonas straminea |