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Literature summary extracted from

  • Kataoka, K.; Takada, H.; Yoshimura, T.; Furuyoshi, S.; Esaki, N.; Ohshima, T.; Soda, K.
    Site-directed mutagenesis of a hexapeptide segment involved in substrate recognition of phenylalanine dehydrogenase from Thermoactinomyces intermedius (1993), J. Biochem., 114, 69-75.
    View publication on PubMed

Protein Variants

EC Number Protein Variants Comment Organism
1.4.1.20 F124M/V125S/H126I/A127I/A128Y/R129Q the catalytic efficiencies of the mutant enzyme with aliphatic amino acids and aliphatic keto acids as substrates are 0.5% to 2% of that of the wild-type enzyme. The efficiencies for L-Phe and phenylpyruvate decreases to 0.0008% and 0.035% of that of the wild-type enzyme, respectively. Enzyme exists as monomeric or dimeric form, compared to wild-type enzyme which exists as hexameric enzyme form. Thermostability is lowered by mutation Thermoactinomyces intermedius

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.4.1.20 0.06
-
 phenylpyruvate wild-type enzyme Thermoactinomyces intermedius
1.4.1.20 0.09
-
 L-Leu wild-type enzyme Thermoactinomyces intermedius
1.4.1.20 0.09
-
 L-Ile wild-type enzyme Thermoactinomyces intermedius
1.4.1.20 0.11
-
 L-Phe
-
 Thermoactinomyces intermedius
1.4.1.20 0.22
-
 norleucine wild-type enzyme Thermoactinomyces intermedius
1.4.1.20 0.27
-
 L-ethionine wild-type enzyme Thermoactinomyces intermedius
1.4.1.20 0.34
-
 L-Met wild-type enzyme Thermoactinomyces intermedius
1.4.1.20 0.53
-
 L-norvaline wild-type enzyme Thermoactinomyces intermedius
1.4.1.20 2.1
-
 L-norleucine mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q Thermoactinomyces intermedius
1.4.1.20 2.5
-
 2-Oxohexanoate wild-type enzyme Thermoactinomyces intermedius
1.4.1.20 2.8
-
 2-oxo-4-methyl-thiobutanoate wild-type enzyme Thermoactinomyces intermedius
1.4.1.20 4
-
 L-ethionine mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q Thermoactinomyces intermedius
1.4.1.20 4.7
-
 L-Leu mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q Thermoactinomyces intermedius
1.4.1.20 7.5
-
 L-Met mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q Thermoactinomyces intermedius
1.4.1.20 9.1
-
 2-Oxoisohexanoate wild-type enzyme Thermoactinomyces intermedius
1.4.1.20 10
-
 L-norvaline mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q Thermoactinomyces intermedius
1.4.1.20 10
-
 L-Ile mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q Thermoactinomyces intermedius
1.4.1.20 13
-
 2-oxo-4-methylpentanoate wild-type enzyme Thermoactinomyces intermedius
1.4.1.20 23
-
 phenylpyruvate mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q Thermoactinomyces intermedius
1.4.1.20 23
-
 L-Phe mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q Thermoactinomyces intermedius
1.4.1.20 35
-
 2-oxopentanoate wild-type enzyme Thermoactinomyces intermedius
1.4.1.20 47
-
 2-Oxohexanoate mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q Thermoactinomyces intermedius
1.4.1.20 50
-
 2-oxo-4-methylpentanoate mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q Thermoactinomyces intermedius
1.4.1.20 56
-
 2-Oxoisohexanoate mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q Thermoactinomyces intermedius
1.4.1.20 87
-
 2-oxo-4-methylthiobutanoate mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q Thermoactinomyces intermedius
1.4.1.20 107
-
 2-oxopentanoate mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q Thermoactinomyces intermedius

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.4.1.20 41000
-
 1 * 41000, the mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q exists as a monomer or a dimer, which are in an equilibrium, SDS-PAGE Thermoactinomyces intermedius
1.4.1.20 41000
-
 6 * 41000, wild-type enzyme, SDS-PAGE Thermoactinomyces intermedius
1.4.1.20 55000
-
 mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q, monomeric enzyme form, gel filtration Thermoactinomyces intermedius
1.4.1.20 110000
-
 mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q, dimeric enzyme form, gel filtration Thermoactinomyces intermedius

Organism

EC Number Organism UniProt Comment Textmining
1.4.1.20 Thermoactinomyces intermedius
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.4.1.20 mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q Thermoactinomyces intermedius

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.4.1.20 additional information
-
-
 Thermoactinomyces intermedius

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.4.1.20 2-oxo-4-methylpentanoate + NH3 + NADH
-
 Thermoactinomyces intermedius L-Ile + NAD+ + H2O
-
 ?
1.4.1.20 2-oxo-4-methylthiobutanoate + NH3 + NADH
-
 Thermoactinomyces intermedius L-Met + NAD+ + H2O
-
 ?
1.4.1.20 2-oxohexanoate + NH3 + NADH
-
 Thermoactinomyces intermedius 2-aminohexanoate + H2O + NAD+
-
 ?
1.4.1.20 2-oxoisohexanoate + NH3 + NADH
-
 Thermoactinomyces intermedius L-Leu + NAD+ + H2O
-
 ?
1.4.1.20 2-oxopentanoate + NH3 + NADH
-
 Thermoactinomyces intermedius L-norvaline + NAD+ + H2O
-
 ?
1.4.1.20 L-ethionine + H2O + NAD+
-
 Thermoactinomyces intermedius 4-ethylthio-2-oxobutanoate + NH3 + NADH
-
 ?
1.4.1.20 L-Leu + H2O + NAD+
-
 Thermoactinomyces intermedius 3-methyl-2-oxopentanoic acid + NH3 + NADH
-
 ?
1.4.1.20 L-Met + H2O + NAD+
-
 Thermoactinomyces intermedius 2-oxo-4-methylthiobutanoate + NH3 + NADH
-
 ?
1.4.1.20 L-norleucine + H2O + NAD+
-
 Thermoactinomyces intermedius 2-oxohexanoic acid + NADH + NH3
-
 ?
1.4.1.20 L-Phe + H2O + NAD+
-
 Thermoactinomyces intermedius phenylpyruvate + NH3 + NADH
-
 r
1.4.1.20 L-Val + H2O + NAD+
-
 Thermoactinomyces intermedius 3-methyl-2-oxobutanoate + NH3 + NADH
-
 ?

Subunits

EC Number Subunits Comment Organism
1.4.1.20 dimer 1 * 41000, the mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q exists as a monomer or a dimer, which are in an equilibrium, SDS-PAGE Thermoactinomyces intermedius
1.4.1.20 hexamer 6 * 41000, wild-type enzyme, SDS-PAGE Thermoactinomyces intermedius
1.4.1.20 monomer 1 * 41000, the mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q exists as a monomer or a dimer, which are in an equilibrium, SDS-PAGE Thermoactinomyces intermedius

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.4.1.20 50
-
 60 min, mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q loses 50% of its activity Thermoactinomyces intermedius
1.4.1.20 70
-
 60 min, wild-type enzyme is stable Thermoactinomyces intermedius

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.4.1.20 0.1
-
 L-ethionine mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q Thermoactinomyces intermedius
1.4.1.20 0.15
-
 L-norvaline mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q Thermoactinomyces intermedius
1.4.1.20 0.16
-
 L-Met mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q Thermoactinomyces intermedius
1.4.1.20 0.17
-
 L-norleucine mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q Thermoactinomyces intermedius
1.4.1.20 0.21
-
 L-Leu mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q Thermoactinomyces intermedius
1.4.1.20 0.24
-
 L-Ile wild-type enzyme Thermoactinomyces intermedius
1.4.1.20 0.28
-
 L-Ile mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q Thermoactinomyces intermedius
1.4.1.20 0.59
-
 L-norvaline wild-type enzyme Thermoactinomyces intermedius
1.4.1.20 0.75
-
 L-Phe mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q Thermoactinomyces intermedius
1.4.1.20 0.84
-
 L-Met wild-type enzyme Thermoactinomyces intermedius
1.4.1.20 1.1
-
 L-ethionine wild-type enzyme Thermoactinomyces intermedius
1.4.1.20 1.9
-
 L-norleucine wild-type enzyme Thermoactinomyces intermedius
1.4.1.20 3.6
-
 L-Leu wild-type enzyme Thermoactinomyces intermedius
1.4.1.20 26
-
 2-oxo-4-methylpentanoate wild-type enzyme Thermoactinomyces intermedius
1.4.1.20 37
-
 2-oxo-4-methylthiobutanoate wild-type enzyme Thermoactinomyces intermedius
1.4.1.20 45
-
 L-Phe wild-type enzyme Thermoactinomyces intermedius
1.4.1.20 57
-
 2-Oxoisohexanoate wild-type enzyme Thermoactinomyces intermedius
1.4.1.20 79
-
 2-Oxohexanoate wild-type enzyme Thermoactinomyces intermedius
1.4.1.20 82
-
 2-oxopentanoate wild-type enzyme Thermoactinomyces intermedius
1.4.1.20 91
-
 phenylpyruvate wild-type enzyme Thermoactinomyces intermedius

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.4.1.20 9
-
 reductive amination of wilde-type enzyme and mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q Thermoactinomyces intermedius
1.4.1.20 9.7 10.1 mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q, oxidative deamination Thermoactinomyces intermedius
1.4.1.20 11
-
 wild-type enzyme, oxidative deamination Thermoactinomyces intermedius

Cofactor

EC Number Cofactor Comment Organism Structure
1.4.1.20 NAD+
-
 Thermoactinomyces intermedius
1.4.1.20 NADH
-
 Thermoactinomyces intermedius