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Literature summary for 1.4.1.20 extracted from

  • Kataoka, K.; Takada, H.; Tanizawa, K.; Yoshimura, T.; Esaki, N.; Ohshima, T.; Soda, K.
    Construction and characterization of chimeric enzyme consisting of an amino-terminal domain of phenylalanine dehydrogenase and a carboxy-terminal domain of leucine dehydrogenase (1994), J. Biochem., 116, 931-936.
    View publication on PubMed

Application

Application Comment Organism
analysis the chimeric enzyme has a specific activity of 6% of that of the parental phenylalanine dehydrogenase Thermoactinomyces intermedius

Cloned(Commentary)

Cloned (Comment) Organism
chimeric enzyme consisting of the N-terminal domain of Thermoactinomyces intermedius phenylalanine dehydrogenase, containing the substrate-binding region and the C-terminal domain of leucine dehydrogenase from Bacillus stearothermophilus containing the NAD+-binding region, expression in Escherichia coli Thermoactinomyces intermedius

Protein Variants

Protein Variants Comment Organism
additional information construction of a chimeric enzyme consisting of the N-terminal domain of Thermoactinomyces intermedius phenylalanine dehydrogenase, containing the substrate-binding region and the C-terminal domain of leucine dehydrogenase from Bacillus stearothermophilus containing the NAD+-binding region. The chimeric enzyme has a specific activity of 6% of that of the parental phenylalanine dehydrogenase and shows a broad substrate specificity in the oxidative deamination, like phenylalanine dehydrogenase. However, it acts much more effectively than phenylalanine dehydrogenase on Ile and Val Thermoactinomyces intermedius

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.06
-
L-Leu chimeric enzyme Thermoactinomyces intermedius
0.11
-
L-Phe
-
Thermoactinomyces intermedius
0.12
-
L-Phe chimeric enzyme Thermoactinomyces intermedius
0.17
-
NAD+
-
Thermoactinomyces intermedius

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
72000
-
chimeric enzyme consisting of the N-terminal domain of Thermoactinomyces intermedius phenylalanine dehydrogenase, containing the substrate-binding region and the C-terminal domain of leucine dehydrogenase from Bacillus stearothermophilus containing the NAD+-binding region, gel filtration Thermoactinomyces intermedius

Organism

Organism UniProt Comment Textmining
Thermoactinomyces intermedius
-
-
-

Purification (Commentary)

Purification (Comment) Organism
construction of a chimeric enzyme consisting of the N-terminal domain of Thermoactinomyces intermedius phenylalanine dehydrogenase, containing the substrate-binding region and the C-terminal domain of leucine dehydrogenase from Bacillus stearothermophilus containing the NAD+-binding region Thermoactinomyces intermedius

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-oxo-4-methylpentanoate + NH3 + NADH 16% of the activity with phenylpyruvate Thermoactinomyces intermedius L-Ile + NAD+ + H2O
-
?
2-oxo-4-methylthiobutanoate + NH3 + NADH 55% of the activity with phenylpyruvate Thermoactinomyces intermedius L-Met + NAD+ + H2O
-
?
2-oxobutanoate + NH3 + NADH 5.5% of the activity with phenylpyruvate Thermoactinomyces intermedius 2-aminobutanoate + NAD+ + H2O
-
?
2-oxohexanoate + NH3 + NADH 130% of the activity with phenylpyruvate Thermoactinomyces intermedius 2-aminohexanoate + H2O + NAD+
-
?
2-oxoisohexanoate + NH3 + NADH 47% of the activity with phenylpyruvate Thermoactinomyces intermedius L-Leu + NAD+ + H2O
-
?
2-oxopentanoate + NH3 + NADH 37% of the activity with phenylpyruvate Thermoactinomyces intermedius L-norvaline + NAD+ + H2O
-
?
4-hydroxyphenylpyruvate + NH3 + NADH 80% of the activity with phenylpyruvate Thermoactinomyces intermedius L-Tyr + NAD+ + H2O
-
?
alloisoleucine + H2O + NAD+ 26% of the activity with L-Phe Thermoactinomyces intermedius 3-methyl-2-oxopentanoate + NADH + NH3
-
?
L-Met + H2O + NAD+ 2.2% of the activity with L-Phe Thermoactinomyces intermedius 2-oxo-4-methylthiobutanoate + NH3 + NADH
-
?
L-norleucine + H2O + NAD+ 30% of the activity with L-Phe Thermoactinomyces intermedius 2-oxohexanoic acid + NADH + NH3
-
?
L-norvaline + H2O + NAD+ 28% of the activity with L-Phe Thermoactinomyces intermedius 2-oxopentanoate + NH3 + NADH
-
?
L-Phe + H2O + NAD+
-
Thermoactinomyces intermedius phenylpyruvate + NH3 + NADH
-
r
L-Tyr + H2O + NAD+ 40% of the activity with L-Phe Thermoactinomyces intermedius 4-hydroxyphenylpyruvate + NH3 + NADH
-
?
m-fluoro-DL-phenylalanine + H2O + NAD+ as effective as L-Phe Thermoactinomyces intermedius 3-(3-fluorophenyl)-2-oxopropionate + NADH + NH3
-
?
additional information specific activity of the chimeric enzyme is 6% of that of the parental phenylalanine dehydrogenase and shows a broad substrate specificity in the oxidative deamination, like phenylalanine dehydrogenase. However, it acts much more effectively than phenylalanine dehydrogenase on Ile and Val. The parent enzyme and the chimeric enzyme belong to the pro-S specific dehydrogenase Thermoactinomyces intermedius ?
-
?
o-fluoro-DL-phenylalanine + H2O + NAD+ 65% of the activity with L-Phe Thermoactinomyces intermedius 3-(2-fluorophenyl)-2-oxopropionate + NADH + NH3
-
?
p-amino-L-phenylalanine + H2O + NAD+
-
Thermoactinomyces intermedius ?
-
?
p-fluoro-DL-phenylalanine + H2O + NAD+ 118% of the activity with L-Phe Thermoactinomyces intermedius (4-fluorophenyl)-2-oxopropionate + NADH + NH3
-
?

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
54
-
60 min, pH 7.0-9.5, chimeric enzyme is stable Thermoactinomyces intermedius
58
-
pH 7.0-9.5, 60 min, chimeric enzyme, loss of activity above Thermoactinomyces intermedius
65 70 60 min, stable Thermoactinomyces intermedius

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
10.6
-
reductive amination of phenylpyruvate, chimeric enzyme Thermoactinomyces intermedius
10.7 11 oxidative deamination of L-Phe, chimeric enzyme Thermoactinomyces intermedius

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Thermoactinomyces intermedius
NADH
-
Thermoactinomyces intermedius