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Literature summary for 1.4.1.9 extracted from

  • Kataoka, K.; Tanizawa, K.
    Alteration of substrate specificity of leucine dehydrogenase by site-directed mutagenesis (2003), J. Mol. Catal. B, 23, 299-309.
No PubMed abstract available

Protein Variants

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 [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

Organism UniProt Comment Textmining
Geobacillus stearothermophilus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme Geobacillus stearothermophilus

Substrates and Products (Substrate)

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

Synonyms Comment Organism
LeuDH
-
Geobacillus stearothermophilus

Turnover Number [1/s]

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

Cofactor Comment Organism Structure
NAD+
-
Geobacillus stearothermophilus
NADH
-
Geobacillus stearothermophilus