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Literature summary for 4.3.1.24 extracted from

  • Zhang, F.; Huang, N.; Zhou, L.; Cui, W.; Liu, Z.; Zhu, L.; Liu, Y.; Zhou, Z.
    Modulating the pH activity profiles of phenylalanine ammonia lyase from Anabaena variabilis by modification of center-near surface residues (2017), Appl. Biochem. Biotechnol., 183, 699-711 .
    View publication on PubMed

Application

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(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli BL21 (DE3) Trichormus variabilis

Protein Variants

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 [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

Organism UniProt Comment Textmining
Trichormus variabilis
-
-
-
Trichormus variabilis FACHB-82
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Trichormus variabilis

Substrates and Products (Substrate)

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 [°C] Temperature Optimum Maximum [°C] Comment Organism
50
-
wild-type and mutant enzymes E75A, and E75Q Trichormus variabilis

Temperature Stability [°C]

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 [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
-
Trichormus variabilis

pH Range

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 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 [mM/s]

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