Protein Variants | Comment | Organism |
---|---|---|
A113G | mutant enzyme with altered substrate specificity. 17.9fold decrease in turnover number for L-Leu, 1.2fold decrease in turnover-number for L-Ile, 13.8fold increase in turnover number of L-norleucine, 1.7fold decrease in turnover-number for L-norvaline, 3fold decrease in turnover number for alpha-keto-isocaproate, 1.2fold decrease in turnover number for alpha-ketocaproate, 1.3fold increase in turnover number for alpha-ketocaproate, 3.6fold decrease in Km-value for L-Leu, 3.3fold increase in Km-value for L-Ile, 1.1fold decrease in Km-value for L-norleucine, 3.5fold increase in Km-value for L-norvaline, 1.9fold increase in Km-value for alpha-keto-isocaproate, 2.5fold increase in Km-value for alpha-keto-beta-methylvalerate, 2.4fold decrease in Km-value for alpha-ketocaproate, 1.2fold increase in Km-value for NAD+, 1.2fold increase in Km-value for NADH as compared to wild-type enzyme. L-Ethionine and L-Phe are not substrates of the wild-type enzyme but are deaminated by mutant enzyme. Phenylpyruvate is not a substrate of the wild-type enzyme, but is aminated by mutant enzyme | Geobacillus stearothermophilus |
A113G/V291L | mutant enzyme with altered substrate specificity. 67.6fold decrease in turnover number for L-Leu, 20fold decrease in turnover-number for L-Ile, 2.2fold decrease in turnover number of L-norleucine, 44.8fold decrease in turnover-number for L-norvaline, 9.7fold decrease in turnover number for alpha-keto-isocaproate, 7.6fold decrease in turnover number for alpha-ketocaproate, 4.6fold decrease in turnover number for alpha-ketocaproate, 6.9fold increase in Km-value for L-Leu, 13.8fold increase in Km-value for L-Ile, 5.5fold increase in Km-value for L-norleucine, 9fold increase in Km-value for L-norvaline, 34fold increase in Km-value for alpha-keto-isocaproate, 18.2fold increase in Km-value for alpha-keto-beta-methylvalerate, 6fold increase in Km-value for alpha-ketocaproate, 4.4fold increase in Km-value for NAD+, 2fold decrease in Km-value for NADH as compared to wild-type enzyme. L-Ethionine and L-Phe are not substrates of the wild-type enzyme but are deaminated by mutant enzyme. Phenylpyruvate is not a substrate of the wild-type enzyme, but is aminated by mutant enzyme | Geobacillus stearothermophilus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.017 | - |
NADH | mutant enzyme A113A/V291L | Geobacillus stearothermophilus | |
0.035 | - |
NADH | wild-type enzyme | Geobacillus stearothermophilus | |
0.042 | - |
NADH | mutant enzyme A113A | Geobacillus stearothermophilus | |
0.063 | - |
NAD+ | wild-type enzyme | Geobacillus stearothermophilus | |
0.076 | - |
NAD+ | mutant enzyme A113A | Geobacillus stearothermophilus | |
0.28 | - |
NAD+ | mutant enzyme A113A/V291L | Geobacillus stearothermophilus | |
0.88 | - |
alpha-keto-isocaproate | wild-type enzyme | Geobacillus stearothermophilus | |
1.4 | - |
L-Leu | mutant enzyme A113A | Geobacillus stearothermophilus | |
1.7 | - |
alpha-keto-isocaproate | mutant enzyme A113A | Geobacillus stearothermophilus | |
2 | - |
2-Oxohexanoate | mutant enzyme A113A | Geobacillus stearothermophilus | |
2.4 | - |
L-Ile | wild-type enzyme | Geobacillus stearothermophilus | |
3.8 | - |
alpha-keto-beta-methylvalerate | wild-type enzyme | Geobacillus stearothermophilus | |
4.1 | - |
L-norleucine | mutant enzyme A113A | Geobacillus stearothermophilus | |
4.4 | - |
L-norleucine | wild-type enzyme | Geobacillus stearothermophilus | |
4.7 | - |
2-Oxohexanoate | wild-type enzyme | Geobacillus stearothermophilus | |
5.1 | - |
L-Leu | wild-type enzyme | Geobacillus stearothermophilus | |
7.1 | - |
phenylpyruvate | mutant enzyme A113A | Geobacillus stearothermophilus | |
7.8 | - |
L-norvaline | wild-type enzyme | Geobacillus stearothermophilus | |
7.8 | - |
L-Ile | mutant enzyme A113A | Geobacillus stearothermophilus | |
9.4 | - |
alpha-keto-beta-methylvalerate | mutant enzyme A113A | Geobacillus stearothermophilus | |
9.9 | - |
phenylpyruvate | mutant enzyme A113A/V291L | Geobacillus stearothermophilus | |
24 | - |
L-norleucine | mutant enzyme A113A/V291L | Geobacillus stearothermophilus | |
27 | - |
L-norvaline | mutant enzyme A113A | Geobacillus stearothermophilus | |
28 | - |
2-Oxohexanoate | mutant enzyme A113A/V291L | Geobacillus stearothermophilus | |
30 | - |
alpha-keto-isocaproate | mutant enzyme A113A/V291L | Geobacillus stearothermophilus | |
31 | - |
L-Phe | mutant enzyme A113A | Geobacillus stearothermophilus | |
33 | - |
L-Ile | mutant enzyme A113A/V291L | Geobacillus stearothermophilus | |
35 | - |
L-Leu | mutant enzyme A113A/V291L | Geobacillus stearothermophilus | |
66 | - |
L-Phe | mutant enzyme A113A/V291L | Geobacillus stearothermophilus | |
69 | - |
alpha-keto-beta-methylvalerate | mutant enzyme A113A/V291L | Geobacillus stearothermophilus | |
70 | - |
L-norvaline | mutant enzyme A113A/V291L | Geobacillus stearothermophilus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Geobacillus stearothermophilus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant enzyme | Geobacillus stearothermophilus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-keto-beta-methylvalerate + NH3 + NADH | as active as 2-ketoisocaproate, wild-type enzyme | Geobacillus stearothermophilus | ? + H2O + NAD+ | - |
? | |
2-keto-gamma-methylthiobutanoate + NH3 + NADH | 15% of the activity with 2-ketoisocaproate, wild-type enzyme | Geobacillus stearothermophilus | L-Met + H2O + NAD+ | - |
? | |
2-ketobutyrate + NH3 + NADH | 47% of the activity with 2-ketoisocaproate, wild-type enzyme | Geobacillus stearothermophilus | L-2-aminobutyrate + H2O + NAD+ | - |
? | |
2-ketoisocaproate + NH3 + NADH | - |
Geobacillus stearothermophilus | L-Ile + H2O + NAD+ | - |
? | |
2-ketovalerate + NH3 + NADH | 86% of the activity with 2-ketoisocaproate, wild-type enzyme | Geobacillus stearothermophilus | L-norvaline + H2O + NAD+ | - |
? | |
2-oxohexanoate + NH3 + NADH + H+ | - |
Geobacillus stearothermophilus | L-norleucine + H2O + NAD+ | - |
r | |
L-Ile + H2O + NAD+ | 54% of the activity with L-Leu, wild-type enzyme | Geobacillus stearothermophilus | 3-methyl-2-oxopentanoate + NH3 + NADH | - |
? | |
L-Leu + H2O + NAD+ | - |
Geobacillus stearothermophilus | 4-methyl-2-oxopentanoate + NH3 + NADH | - |
? | |
L-Met + H2O + NAD+ | 0.7% the activity with L-Leu, wild-type enzyme | Geobacillus stearothermophilus | 4-methylthio-2-oxobutyrate + NH3 + NADH + H+ | - |
? | |
L-norleucine + H2O + NAD+ | 14% the activity with L-Leu, wild-type enzyme | Geobacillus stearothermophilus | ? + NH3 + NADH | - |
? | |
L-norvaline + H2O + NAD+ | 56% the activity with L-Leu, wild-type enzyme | Geobacillus stearothermophilus | 2-ketovalerate + NH3 + NADH | - |
? | |
L-Phe + H2O + NAD+ | no activity of wild-type enzyme activity with mutant enzymes A113G, A113G/V291L | Geobacillus stearothermophilus | phenylpyruvate + NH3 + NADH | - |
? | |
L-Val + H2O + NAD+ | 39% of the activity with L-Leu, wild-type enzyme | Geobacillus stearothermophilus | 3-methyl-2-oxobutanoate + NH3 + NADH | - |
? | |
phenylpyruvate + NH3 + NADH | 15% of the activity with 2-ketoisocaproate, wild-type enzyme | Geobacillus stearothermophilus | L-Phe + H2O + NAD+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
LeuDH | - |
Geobacillus stearothermophilus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.29 | - |
L-norvaline | mutant enzyme A113A/V291L | Geobacillus stearothermophilus | |
0.6 | - |
L-norleucine | mutant enzyme A113A/V291L | Geobacillus stearothermophilus | |
0.74 | - |
L-Leu | mutant enzyme A113A/V291L | Geobacillus stearothermophilus | |
0.8 | - |
L-Phe | mutant enzyme A113A | Geobacillus stearothermophilus | |
1.3 | - |
L-norleucine | wild-type enzyme | Geobacillus stearothermophilus | |
1.4 | - |
L-Phe | mutant enzyme A113A/V291L | Geobacillus stearothermophilus | |
1.4 | - |
L-Ile | mutant enzyme A113A/V291L | Geobacillus stearothermophilus | |
2 | 8 | L-Ile | wild-type enzyme | Geobacillus stearothermophilus | |
2.8 | - |
L-Leu | mutant enzyme A113A | Geobacillus stearothermophilus | |
7.7 | - |
L-norvaline | mutant enzyme A113A | Geobacillus stearothermophilus | |
9.9 | - |
phenylpyruvate | mutant enzyme A113A/V291L | Geobacillus stearothermophilus | |
13 | - |
L-norvaline | wild-type enzyme | Geobacillus stearothermophilus | |
18 | - |
L-norleucine | mutant enzyme A113A | Geobacillus stearothermophilus | |
22 | - |
phenylpyruvate | mutant enzyme A113A | Geobacillus stearothermophilus | |
23 | - |
L-Ile | mutant enzyme A113A | Geobacillus stearothermophilus | |
24 | - |
2-Oxohexanoate | mutant enzyme A113A/V291L | Geobacillus stearothermophilus | |
29 | - |
alpha-keto-isocaproate | mutant enzyme A113A/V291L | Geobacillus stearothermophilus | |
37 | - |
alpha-keto-beta-methylvalerate | mutant enzyme A113A/V291L | Geobacillus stearothermophilus | |
50 | - |
L-Leu | wild-type enzyme | Geobacillus stearothermophilus | |
94 | - |
alpha-keto-isocaproate | mutant enzyme A113A | Geobacillus stearothermophilus | |
110 | - |
2-Oxohexanoate | wild-type enzyme | Geobacillus stearothermophilus | |
140 | - |
2-Oxohexanoate | mutant enzyme A113A | Geobacillus stearothermophilus | |
235 | - |
alpha-keto-beta-methylvalerate | mutant enzyme A113A | Geobacillus stearothermophilus | |
280 | - |
alpha-keto-beta-methylvalerate | wild-type enzyme | Geobacillus stearothermophilus | |
280 | - |
alpha-keto-isocaproate | wild-type enzyme | Geobacillus stearothermophilus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Geobacillus stearothermophilus | |
NADH | - |
Geobacillus stearothermophilus |