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 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 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 | UniProt | Comment | Textmining |
---|---|---|---|
Thermoactinomyces intermedius | - |
- |
- |
Purification (Comment) | Organism |
---|---|
mutant enzyme F124M/V125S/H126I/A127I/A128Y/R129Q | Thermoactinomyces intermedius |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
- |
Thermoactinomyces intermedius |
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 | 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 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 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 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 | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Thermoactinomyces intermedius | |
NADH | - |
Thermoactinomyces intermedius |