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Literature summary for 1.4.1.9 extracted from

  • Zhou, J.; Wang, Y.; Chen, J.; Xu, M.; Yang, T.; Zheng, J.; Zhang, X.; Rao, Z.
    Rational engineering of Bacillus cereus leucine dehydrogenase towards alpha-keto acid reduction for improving unnatural amino acid production (2019), Biotechnol. J., 14, e1800253 .
    View publication on PubMed
Search for more literature for 1.4.1.9

Protein Variants

Protein Variants Comment Organism
E116V 123% of wild-type specific activity. Variant is associated with the presence of more hydrophobic tunnels that allow easy substrate diffusion Bacillus cereus
T45M an additional hydrogen bond in beta5 fold is formed, which results in a more rigid beta5 fold leading to better stability Bacillus cereus
T45M/E116V mutant displays higher productivities, melting temperature and half-lives of thermal inactivation are increased compared to wild type Bacillus cereus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.07
-
 NADH mutant E116V, pH 9.5, 35°C Bacillus cereus
0.09
-
 NADH mutant T45M/E116V, pH 9.5, 35°C Bacillus cereus
0.17
-
 NADH wild-type, pH 9.5, 35°C Bacillus cereus
0.22
-
 NADH mutant T45M, pH 9.5, 35°C Bacillus cereus
0.38
-
 4-methyl-2-oxopentanoate mutant E116V, pH 9.5, 35°C Bacillus cereus
0.41
-
 2-oxopentanoate mutant T45M/E116V, pH 9.5, 35°C Bacillus cereus
0.705
-
 2-oxobutanoate mutant E116V, pH 9.5, 35°C Bacillus cereus
0.73
-
 2-oxo-3-methylbutanoate mutant E116V, pH 9.5, 35°C Bacillus cereus
0.75
-
 oxo(phenyl)acetate mutant E116V, pH 9.5, 35°C Bacillus cereus
0.89
-
 4-methyl-2-oxopentanoate mutant T45M/E116V, pH 9.5, 35°C Bacillus cereus
0.93
-
 2-oxo-3-methylbutanoate mutant T45M/E116V, pH 9.5, 35°C Bacillus cereus
1.12
-
 oxo(phenyl)acetate mutant T45M/E116V, pH 9.5, 35°C Bacillus cereus
1.14
-
 2-oxopentanoate mutant E116V, pH 9.5, 35°C Bacillus cereus
1.15
-
 trimethylpyruvate mutant T45M, pH 9.5, 35°C Bacillus cereus
1.41
-
 4-methyl-2-oxopentanoate wild-type, pH 9.5, 35°C Bacillus cereus
1.43
-
 2-oxopentanoate wild-type, pH 9.5, 35°C Bacillus cereus
1.7
-
 2-oxobutanoate mutant T45M, pH 9.5, 35°C Bacillus cereus
1.71
-
 trimethylpyruvate mutant E116V, pH 9.5, 35°C Bacillus cereus
1.72
-
 4-methyl-2-oxopentanoate mutant T45M, pH 9.5, 35°C Bacillus cereus
1.89
-
 2-oxobutanoate wild-type, pH 9.5, 35°C Bacillus cereus
1.99
-
 2-oxopentanoate mutant T45M, pH 9.5, 35°C Bacillus cereus
1.99
-
 oxo(phenyl)acetate mutant T45M, pH 9.5, 35°C Bacillus cereus
2.47
-
 2-oxo-3-methylbutanoate wild-type, pH 9.5, 35°C Bacillus cereus
2.54
-
 2-oxo-3-methylbutanoate mutant T45M, pH 9.5, 35°C Bacillus cereus
2.65
-
 2-oxobutanoate mutant T45M/E116V, pH 9.5, 35°C Bacillus cereus
2.65
-
 trimethylpyruvate mutant T45M/E116V, pH 9.5, 35°C Bacillus cereus
3.7 5 trimethylpyruvate wild-type, pH 9.5, 35°C Bacillus cereus
9.38
-
 oxo(phenyl)acetate wild-type, pH 9.5, 35°C Bacillus cereus

Organism

Organism UniProt Comment Textmining
Bacillus cereus P0A392
-
-
Bacillus cereus DSM 31 P0A392
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-oxo-3-methylbutanoate + NH3 + NADH + H+
-
 Bacillus cereus L-2-amino-3-methylbutanoate + H2O + NAD+
-
 ?
2-oxo-3-methylbutanoate + NH3 + NADH + H+
-
 Bacillus cereus DSM 31 L-2-amino-3-methylbutanoate + H2O + NAD+
-
 ?
2-oxobutanoate + NH3 + NADH + H+
-
 Bacillus cereus L-2-aminobutanoate + H2O + NAD+
-
 ?
2-oxobutanoate + NH3 + NADH + H+
-
 Bacillus cereus DSM 31 L-2-aminobutanoate + H2O + NAD+
-
 ?
2-oxopentanoate + NH3 + NADH + H+
-
 Bacillus cereus L-2-aminopentanoate + H2O + NAD+
-
 ?
2-oxopentanoate + NH3 + NADH + H+
-
 Bacillus cereus DSM 31 L-2-aminopentanoate + H2O + NAD+
-
 ?
4-methyl-2-oxopentanoate + NH3 + NADH + H+
-
 Bacillus cereus L-4-methyl-2-aminopentanoate + H2O + NAD+
-
 ?
4-methyl-2-oxopentanoate + NH3 + NADH + H+
-
 Bacillus cereus DSM 31 L-4-methyl-2-aminopentanoate + H2O + NAD+
-
 ?
oxo(phenyl)acetate + NH3 + NADH + H+
-
 Bacillus cereus amino(phenyl)acetate + H2O + NAD+
-
 ?
trimethylpyruvate + NH3 + NADH + H+
-
 Bacillus cereus L-tert-leucine + H2O + NAD+
-
 ?
trimethylpyruvate + NH3 + NADH + H+
-
 Bacillus cereus DSM 31 L-tert-leucine + H2O + NAD+
-
 ?

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
48.4
-
 melting temperature of wild-type Bacillus cereus
58
-
 melting temperature of mutant T45M Bacillus cereus
62.2
-
 melting temperature of mutant E116V Bacillus cereus
62.8
-
 melting temperature of mutant T45M/E116V Bacillus cereus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.006
-
 trimethylpyruvate mutant T45M, pH 9.5, 35°C Bacillus cereus
0.016
-
 trimethylpyruvate wild-type, pH 9.5, 35°C Bacillus cereus
0.033
-
 trimethylpyruvate mutant E116V, pH 9.5, 35°C Bacillus cereus
0.036
-
 trimethylpyruvate mutant T45M/E116V, pH 9.5, 35°C Bacillus cereus
0.06
-
 oxo(phenyl)acetate mutant T45M, pH 9.5, 35°C Bacillus cereus
0.101
-
 oxo(phenyl)acetate mutant E116V, pH 9.5, 35°C Bacillus cereus
0.129
-
 2-oxopentanoate mutant T45M/E116V, pH 9.5, 35°C Bacillus cereus
0.149
-
 2-oxobutanoate wild-type, pH 9.5, 35°C Bacillus cereus
0.168
-
 2-oxobutanoate mutant E116V, pH 9.5, 35°C Bacillus cereus
0.174
-
 2-oxobutanoate mutant T45M, pH 9.5, 35°C Bacillus cereus
0.182
-
 oxo(phenyl)acetate wild-type, pH 9.5, 35°C Bacillus cereus
0.214
-
 oxo(phenyl)acetate mutant T45M/E116V, pH 9.5, 35°C Bacillus cereus
0.284
-
 2-oxobutanoate mutant T45M/E116V, pH 9.5, 35°C Bacillus cereus
0.286
-
 NADH mutant T45M/E116V, pH 9.5, 35°C Bacillus cereus
0.332
-
 2-oxopentanoate wild-type, pH 9.5, 35°C Bacillus cereus
0.382
-
 2-oxo-3-methylbutanoate mutant T45M/E116V, pH 9.5, 35°C Bacillus cereus
0.384
-
 2-oxopentanoate mutant E116V, pH 9.5, 35°C Bacillus cereus
0.385
-
 4-methyl-2-oxopentanoate mutant T45M/E116V, pH 9.5, 35°C Bacillus cereus
0.467
-
 4-methyl-2-oxopentanoate wild-type, pH 9.5, 35°C Bacillus cereus
0.478
-
 2-oxo-3-methylbutanoate wild-type, pH 9.5, 35°C Bacillus cereus
0.532
-
 4-methyl-2-oxopentanoate mutant T45M, pH 9.5, 35°C Bacillus cereus
0.543
-
 4-methyl-2-oxopentanoate mutant E116V, pH 9.5, 35°C Bacillus cereus
0.544
-
 NADH wild-type, pH 9.5, 35°C Bacillus cereus
0.552
-
 NADH mutant T45M, pH 9.5, 35°C Bacillus cereus
0.588
-
 2-oxopentanoate mutant T45M, pH 9.5, 35°C Bacillus cereus
0.659
-
 2-oxo-3-methylbutanoate mutant E116V, pH 9.5, 35°C Bacillus cereus
0.702
-
 2-oxo-3-methylbutanoate mutant T45M, pH 9.5, 35°C Bacillus cereus
1.3
-
 NADH mutant E116V, pH 9.5, 35°C Bacillus cereus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.004
-
 trimethylpyruvate wild-type, pH 9.5, 35°C Bacillus cereus
0.005
-
 trimethylpyruvate mutant T45M, pH 9.5, 35°C Bacillus cereus
0.014
-
 trimethylpyruvate mutant T45M/E116V, pH 9.5, 35°C Bacillus cereus
0.019
-
 trimethylpyruvate mutant E116V, pH 9.5, 35°C Bacillus cereus
0.019
-
 oxo(phenyl)acetate wild-type, pH 9.5, 35°C Bacillus cereus
0.03
-
 oxo(phenyl)acetate mutant T45M, pH 9.5, 35°C Bacillus cereus
0.079
-
 2-oxobutanoate wild-type, pH 9.5, 35°C Bacillus cereus
0.102
-
 2-oxobutanoate mutant T45M, pH 9.5, 35°C Bacillus cereus
0.107
-
 2-oxobutanoate mutant T45M/E116V, pH 9.5, 35°C Bacillus cereus
0.134
-
 oxo(phenyl)acetate mutant E116V, pH 9.5, 35°C Bacillus cereus
0.191
-
 oxo(phenyl)acetate mutant T45M/E116V, pH 9.5, 35°C Bacillus cereus
0.194
-
 2-oxo-3-methylbutanoate wild-type, pH 9.5, 35°C Bacillus cereus
0.232
-
 2-oxopentanoate wild-type, pH 9.5, 35°C Bacillus cereus
0.238
-
 2-oxobutanoate mutant E116V, pH 9.5, 35°C Bacillus cereus
0.276
-
 2-oxo-3-methylbutanoate mutant T45M, pH 9.5, 35°C Bacillus cereus
0.295
-
 2-oxopentanoate mutant T45M, pH 9.5, 35°C Bacillus cereus
0.309
-
 4-methyl-2-oxopentanoate mutant T45M, pH 9.5, 35°C Bacillus cereus
0.315
-
 2-oxopentanoate mutant T45M/E116V, pH 9.5, 35°C Bacillus cereus
0.332
-
 4-methyl-2-oxopentanoate wild-type, pH 9.5, 35°C Bacillus cereus
0.337
-
 2-oxopentanoate mutant E116V, pH 9.5, 35°C Bacillus cereus
0.413
-
 2-oxo-3-methylbutanoate mutant T45M/E116V, pH 9.5, 35°C Bacillus cereus
0.433
-
 4-methyl-2-oxopentanoate mutant T45M/E116V, pH 9.5, 35°C Bacillus cereus
0.902
-
 2-oxo-3-methylbutanoate mutant E116V, pH 9.5, 35°C Bacillus cereus
1.44
-
 4-methyl-2-oxopentanoate mutant E116V, pH 9.5, 35°C Bacillus cereus
2.46
-
 NADH mutant T45M, pH 9.5, 35°C Bacillus cereus
3.18
-
 NADH mutant T45M/E116V, pH 9.5, 35°C Bacillus cereus
3.18
-
 NADH wild-type, pH 9.5, 35°C Bacillus cereus
18.7
-
 NADH mutant E116V, pH 9.5, 35°C Bacillus cereus