Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.4.1.20 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
Search for more literature for 1.4.1.20

Protein Variants

Protein Variants Comment Organism
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]

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

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
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
41000
-
 6 * 41000, wild-type enzyme, SDS-PAGE Thermoactinomyces intermedius
55000
-
 mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q, monomeric enzyme form, gel filtration Thermoactinomyces intermedius
110000
-
 mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q, dimeric enzyme form, gel filtration Thermoactinomyces intermedius

Organism

Organism UniProt Comment Textmining
Thermoactinomyces intermedius
-
-
-

Purification (Commentary)

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

Specific Activity [micromol/min/mg]

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

Substrates and Products (Substrate)

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

Subunits

Subunits Comment Organism
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
hexamer 6 * 41000, wild-type enzyme, SDS-PAGE Thermoactinomyces intermedius
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]

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

Turnover Number [1/s]

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

pH Optimum

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

Cofactor

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