Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3)pLysS cells | Pseudomonas putida |
Protein Variants | Comment | Organism |
---|---|---|
A460Y | the mutant shows significantly improved catalytic activity toward pyruvate | Pseudomonas putida |
T377L | the mutant shows significantly improved catalytic activity toward pyruvate | Pseudomonas putida |
T377L/A460Y | the double mutant shows significantly improved catalytic activity toward pyruvate (11000fold improvement in the ratio between pyruvate and benzoylformate utilization) | Pseudomonas putida |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
(R)-mandelate | substrate analogue inhibitor | Pseudomonas putida |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.04 | - |
benzylformate | mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
0.1 | - |
2-ketopentanoate | mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
0.15 | - |
2-ketohexanoate | mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
0.27 | - |
benzylformate | wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
0.28 | - |
2-ketopentanoate | mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
0.28 | - |
2-ketohexanoate | mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
0.42 | - |
2-ketobutanoate | mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
0.45 | - |
benzylformate | mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
1.1 | - |
2-ketopentanoate | mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
1.3 | - |
2-ketobutanoate | mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
1.3 | - |
2-ketohexanoate | mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
1.5 | - |
benzylformate | mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
2 | - |
pyruvate | mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
4.1 | - |
2-ketohexanoate | wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
4.3 | - |
2-ketobutanoate | mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
4.8 | - |
pyruvate | mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
6 | - |
2-ketopentanoate | wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
7.5 | - |
2-ketobutanoate | wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
21 | - |
pyruvate | mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas putida | P20906 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Pseudomonas putida |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-ketohexanoate | - |
Pseudomonas putida | valeraldehyde + CO2 | - |
? | |
2-ketopentanoate | - |
Pseudomonas putida | butyraldehyde + CO2 | - |
? | |
2-oxobutanoate | - |
Pseudomonas putida | propionaldehyde + CO2 | - |
? | |
benzylformate | - |
Pseudomonas putida | benzaldehyde + CO2 | - |
? | |
pyruvate | pyruvate is no substrate for the wild type enzyme | Pseudomonas putida | acetaldehyde + CO2 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
BFDC | - |
Pseudomonas putida |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.81 | - |
2-ketopentanoate | mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
1.1 | - |
2-ketohexanoate | mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
1.2 | - |
2-ketohexanoate | mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
1.4 | - |
pyruvate | mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
1.7 | - |
benzylformate | mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
2 | - |
pyruvate | mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
3.3 | - |
2-ketopentanoate | mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
3.6 | - |
pyruvate | mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
4 | - |
2-ketobutanoate | wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
4.4 | - |
2-ketohexanoate | mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
5.3 | - |
2-ketohexanoate | wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
5.8 | - |
2-ketobutanoate | mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
6 | - |
2-ketobutanoate | mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
6.2 | - |
2-ketopentanoate | mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
7.5 | - |
2-ketobutanoate | mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
11 | - |
2-ketopentanoate | wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
25 | - |
benzylformate | mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
110 | - |
benzylformate | mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
320 | - |
benzylformate | wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
thiamine diphosphate | - |
Pseudomonas putida |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.012 | - |
pyruvate | wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
0.065 | - |
pyruvate | mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
0.42 | - |
pyruvate | mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
0.5 | - |
2-ketobutanoate | wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
0.86 | - |
2-ketohexanoate | mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
1.3 | - |
2-ketohexanoate | wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
1.7 | - |
2-ketobutanoate | mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
1.8 | - |
pyruvate | mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
1.8 | - |
2-ketopentanoate | wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
3 | - |
2-ketopentanoate | mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
4.6 | - |
2-ketobutanoate | mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
8 | - |
2-ketohexanoate | mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
8.1 | - |
2-ketopentanoate | mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
14 | - |
2-ketobutanoate | mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
16 | - |
2-ketohexanoate | mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
16 | - |
benzylformate | mutant enzyme T377L/A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
22 | - |
2-ketopentanoate | mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
43 | - |
benzylformate | mutant enzyme A460Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
244 | - |
benzylformate | mutant enzyme T377L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida | |
1180 | - |
benzylformate | wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C | Pseudomonas putida |