Protein Variants | Comment | Organism |
---|---|---|
K80A | markedly reduced activity in oxidative deamination, nearly 90% of the wild-type activity in reductive amination. Km-value for 2-oxoisohexanoate is 11fold higher than that of the wild-type enzyme, Km-value for L-Leu is lower than that of the wild-type enzyme | Geobacillus stearothermophilus |
K80Q | markedly reduced activity in oxidative deamination. Km-value for 2-oxoisohexanoate is 28fold higher than that of the wild-type enzyme, Km-value for L-Leu is about 3times larger than that of the wild-type enzyme | Geobacillus stearothermophilus |
K80R | markedly reduced activity in oxidative deamination, 0.6% of the wild-type activity in reductive amination, Km-value for L-Leu is lower than that of the wild-type enzyme | Geobacillus stearothermophilus |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
4-methyl-2-pentanone | competitive inhibition of wild-type enzyme, noncompetitive inhibition of mutant enzyme K80A | Geobacillus stearothermophilus | |
4-methylpentanoate | - |
Geobacillus stearothermophilus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.022 | - |
NADH | mutant enzyme K80A | Geobacillus stearothermophilus | |
0.034 | - |
NAD+ | mutant enzyme K80Q | Geobacillus stearothermophilus | |
0.035 | - |
NADH | wild-type enzyme | Geobacillus stearothermophilus | |
0.037 | - |
NADH | mutant enzyme K80R | Geobacillus stearothermophilus | |
0.045 | - |
NADH | mutant enzyme K80Q | Geobacillus stearothermophilus | |
0.063 | - |
NAD+ | wild-type enzyme | Geobacillus stearothermophilus | |
0.074 | - |
NAD+ | mutant enzyme K80R | Geobacillus stearothermophilus | |
0.14 | - |
NAD+ | mutant enzyme K80A | Geobacillus stearothermophilus | |
0.88 | - |
2-Oxoisohexanoate | wild-type enzyme | Geobacillus stearothermophilus | |
0.99 | - |
2-Oxoisohexanoate | mutant enzyme K80R | Geobacillus stearothermophilus | |
2.8 | - |
NAD+ | mutant enzyme K80R | Geobacillus stearothermophilus | |
3.7 | - |
L-Leu | mutant enzyme K80A | Geobacillus stearothermophilus | |
5.1 | - |
L-Leu | wild-type enzyme | Geobacillus stearothermophilus | |
9.8 | - |
2-Oxoisohexanoate | mutant enzyme K80A | Geobacillus stearothermophilus | |
17 | - |
NAD+ | mutant enzyme K80Q | Geobacillus stearothermophilus | |
25 | - |
2-Oxoisohexanoate | mutant enzyme K80Q | Geobacillus stearothermophilus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Geobacillus stearothermophilus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
- |
Geobacillus stearothermophilus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-Leu + H2O + NAD+ | - |
Geobacillus stearothermophilus | 4-methyl-2-oxopentanoate + NH3 + NADH | - |
r |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
75 | - |
unfolding temperature of mutant enzyme K80A | Geobacillus stearothermophilus |
79 | - |
unfolding temperature of mutant enzyme K80R and K80Q | Geobacillus stearothermophilus |
80 | - |
unfolding temperature of the wild-type enzyme | Geobacillus stearothermophilus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | turnover numbers of wild-type enzyme and mutant enzymes | Geobacillus stearothermophilus |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
7 | 11.2 | 25°C, 30 min, wild-type enzyme and mutant enzymes K80A, K80R and K80Q | Geobacillus stearothermophilus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Geobacillus stearothermophilus | |
NADH | - |
Geobacillus stearothermophilus |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
25 | - |
4-methylpentanoate | wild-type enzyme | Geobacillus stearothermophilus | |
33 | - |
4-methylpentanoate | mutant enzyme K80A | Geobacillus stearothermophilus | |
140 | - |
4-methyl-2-pentanone | wild-type enzyme | Geobacillus stearothermophilus |