Application | Comment | Organism |
---|---|---|
medicine | the shift of the pH-optimum from pH 8.5 for the wild-type enzyme to pH 7.5 with 30% higher specific activity than that of the wild-type enzyme, the prolonged half-life of the mutant enzyme at 70°C, the higher resistance to a low pH of 3.5 and protease make the mutant enzyme E75L a candidate for oral medicine of phenylketonuria | Trichormus variabilis |
pharmacology | the shift of the pH-optimum from pH 8.5 for the wild-type enzyme to pH 7.5 with 30% higher specific activity than that of the wild-type enzyme, the prolonged half-life of the mutant enzyme at 70°C, the higher resistance to a low pH of 3.5 and protease make the mutant enzyme E75L a candidate for oral medicine of phenylketonuria | Trichormus variabilis |
Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli BL21 (DE3) | Trichormus variabilis |
Protein Variants | Comment | Organism |
---|---|---|
E75A | shift of the pH optimum from pH 8.5 for the wild-type enzyme to pH 7.5 with 35% higher specific activity than that of the wild-type enzyme | Trichormus variabilis |
E75L | shift of the pH optimum from pH 8.5 for the wild-type enzyme to pH 7.5 with 30% higher specific activity than that of the wild-type enzyme. The half-life of the mutant enzyme at 70°C is prolonged to 190 min from 130 min of the wild-type enzyme. The higher resistance to a low pH of 3.5 and protease make the mutant enzyme a candidate for oral medicine of phenylketonuria | Trichormus variabilis |
E75Q | shift of the pH optimum from pH 8.5 for the wild-type enzyme to pH 7.5 with 24% higher specific activity than that of the wild-type enzyme. The half-life of the mutant enzyme at 70°C is prolonged to 180 min from 130 min of the wild-type enzyme | Trichormus variabilis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.077 | - |
L-phenylalanine | 37°C, pH 7.5, wild-type enzyme | Trichormus variabilis | |
0.078 | - |
L-phenylalanine | 37°C, pH 7.5, mutant enzyme E75L | Trichormus variabilis | |
0.099 | - |
L-phenylalanine | 37°C, pH 7.5, mutant enzyme E75A | Trichormus variabilis | |
0.101 | - |
L-phenylalanine | 37°C, pH 7.5, mutant enzyme E75Q | Trichormus variabilis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Trichormus variabilis | - |
- |
- |
Trichormus variabilis FACHB-82 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
- |
Trichormus variabilis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-phenylalanine | - |
Trichormus variabilis | trans-cinnamate + NH3 | - |
? | |
L-phenylalanine | - |
Trichormus variabilis FACHB-82 | trans-cinnamate + NH3 | - |
? |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
50 | - |
wild-type and mutant enzymes E75A, and E75Q | Trichormus variabilis |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
70 | - |
half-life: 130 min (wild-type enzyme), 190 min (mutant enzyme E75L), 180 min (mutant enzyme E75Q) | Trichormus variabilis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.599 | - |
L-phenylalanine | 37°C, pH 7.5, wild-type enzyme | Trichormus variabilis | |
1.84 | - |
L-phenylalanine | 37°C, pH 7.5, mutant enzyme E75L | Trichormus variabilis | |
2.399 | - |
L-phenylalanine | 37°C, pH 7.5, mutant enzyme E75A | Trichormus variabilis | |
2.478 | - |
L-phenylalanine | 37°C, pH 7.5, mutant enzyme E75Q | Trichormus variabilis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | - |
- |
Trichormus variabilis |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
7.3 | 8.5 | the enzyme maintains 70% of its highest activity (at pH 8.5) when pH decreases to 7.3-7.4 | Trichormus variabilis |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
after incubation for 12 h in pH 7.5, mutant enzyme E75Q almost kept its initial activity, while the wild-type enzyme loses 15% initial activity | Trichormus variabilis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
20.77 | - |
L-phenylalanine | 37°C, pH 7.5, wild-type enzyme | Trichormus variabilis | |
23.58 | - |
L-phenylalanine | 37°C, pH 7.5, mutant enzyme E75L | Trichormus variabilis | |
24.26 | - |
L-phenylalanine | 37°C, pH 7.5, mutant enzyme E75A | Trichormus variabilis | |
24.46 | - |
L-phenylalanine | 37°C, pH 7.5, mutant enzyme E75Q | Trichormus variabilis |