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Literature summary extracted from

  • Meng, X.; Yang, L.; Liu, Y.; Wang, H.; Shen, Y.; Wei, D.
    Identification and rational engineering of a high substrate-tolerant leucine dehydrogenase effective for the synthesis of L-tert-leucine (2021), ChemCatChem, 13, 3340-3349 .
No PubMed abstract available

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.4.1.9 expression in Escherichia coli Roseibium aggregatum

Protein Variants

EC Number Protein Variants Comment Organism
1.4.1.9 D153 N/H191 N 25fold improved affinity for NADH and with 50fold enhanced catalytic efficiency Roseibium aggregatum
1.4.1.9 D153N affinity for NADH is improved, 11.3fold decrease in Km value for NADH Roseibium aggregatum
1.4.1.9 H191N affinity for NADH is improved, 6.0fold decrease in Km value for NADH Roseibium aggregatum
1.4.1.9 P130V affinity for NADH is improved, 8.6fold decrease in Km value for NADH Roseibium aggregatum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.4.1.9 0.02
-
 NADH mutant D153N/H191N, pH 8, 30°C Roseibium aggregatum
1.4.1.9 0.04
-
 NADH mutant D153N, pH 8, 30°C Roseibium aggregatum
1.4.1.9 0.05
-
 NADH mutant P130V, pH 8, 30°C Roseibium aggregatum
1.4.1.9 0.07
-
 NADH mutant H191N, pH 8, 30°C Roseibium aggregatum
1.4.1.9 0.078
-
 NAD+ wild-type, pH 8, 30°C Roseibium aggregatum
1.4.1.9 0.09
-
 trimethylpyruvate mutant H191N, pH 8, 30°C Roseibium aggregatum
1.4.1.9 0.096
-
 trimethylpyruvate wild-type, pH 8, 30°C Roseibium aggregatum
1.4.1.9 0.1
-
 trimethylpyruvate mutant D153N/H191N, pH 8, 30°C Roseibium aggregatum
1.4.1.9 0.12
-
 trimethylpyruvate mutant D153N, pH 8, 30°C Roseibium aggregatum
1.4.1.9 0.3
-
 trimethylpyruvate mutant P130V, pH 8, 30°C Roseibium aggregatum
1.4.1.9 0.43
-
 NADH wild-type, pH 8, 30°C Roseibium aggregatum
1.4.1.9 5.05
-
 L-tert-leucine wild-type, pH 8, 30°C Roseibium aggregatum

Organism

EC Number Organism UniProt Comment Textmining
1.4.1.9 Roseibium aggregatum
-
-
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.4.1.9 556.6
-
 substrate trimethylpyruvate, wild-type, pH 8, 30°C Roseibium aggregatum
1.4.1.9 1410
-
 substrate oxo(phenyl)acetate, wild-type, pH 8, 30°C Roseibium aggregatum
1.4.1.9 1710
-
 substrate oxo(phenyl)acetate, mutant D153N/H191N, pH 8, 30°C Roseibium aggregatum
1.4.1.9 1743
-
 substrate 2-oxopropanoate, wild-type, pH 8, 30°C Roseibium aggregatum
1.4.1.9 2217
-
 substrate 2-oxopropanoate, mutant D153N/H191N, pH 8, 30°C Roseibium aggregatum
1.4.1.9 9046
-
 substrate 4-methyl-2-oxopentanoate, wild-type, pH 8, 30°C Roseibium aggregatum
1.4.1.9 13099
-
 substrate 2-aminopentanoate, wild-type, pH 8, 30°C Roseibium aggregatum
1.4.1.9 15871
-
 substrate 2-oxobutanoate, wild-type, pH 8, 30°C Roseibium aggregatum
1.4.1.9 19609
-
 substrate 4-methyl-2-oxopentanoate, mutant D153N/H191N, pH 8, 30°C Roseibium aggregatum
1.4.1.9 23654
-
 substrate 2-aminopentanoate, mutant D153N/H191N, pH 8, 30°C Roseibium aggregatum
1.4.1.9 31256
-
 substrate 2-oxobutanoate, mutant D153N/H191N, pH 8, 30°C Roseibium aggregatum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.4.1.9 2-oxobutanoate + NH3 + NADH + H+
-
 Roseibium aggregatum L-2-aminobutanoate + H2O + NAD+
-
 ?
1.4.1.9 2-oxopentanoate + NH3 + NADH + H+
-
 Roseibium aggregatum L-2-aminopentanoate + H2O + NAD+
-
 ?
1.4.1.9 2-oxopropanoate + NH3 + NADH + H+
-
 Roseibium aggregatum L-alanine + H2O + NAD+
-
 ?
1.4.1.9 4-methyl-2-oxopentanoate + NH3 + NADH + H+
-
 Roseibium aggregatum L-4-methyl-2-aminopentanoate + H2O + NAD+
-
 ?
1.4.1.9 L-tert-leucine + H2O + NAD+
-
 Roseibium aggregatum trimethylpyruvate + NH3 + NADH + H+
-
 ?
1.4.1.9 oxo(phenyl)acetate + NH3 + NADH + H+
-
 Roseibium aggregatum amino(phenyl)acetate + H2O + NAD+
-
 ?
1.4.1.9 trimethylpyruvate + NH3 + NADH + H+
-
 Roseibium aggregatum L-tert-leucine + H2O + NAD+
-
 ?

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.4.1.9 40
-
-
 Roseibium aggregatum

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.4.1.9 30
-
 half-life 72 h Roseibium aggregatum
1.4.1.9 35
-
 half-life 48 h Roseibium aggregatum
1.4.1.9 40
-
 half-life 20 h Roseibium aggregatum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.4.1.9 33.9
-
 L-tert-leucine wild-type, pH 8, 30°C Roseibium aggregatum
1.4.1.9 347
-
 NADH wild-type, pH 8, 30°C Roseibium aggregatum
1.4.1.9 347
-
 trimethylpyruvate wild-type, pH 8, 30°C Roseibium aggregatum
1.4.1.9 571
-
 NADH mutant D153N, pH 8, 30°C Roseibium aggregatum
1.4.1.9 571
-
 NADH mutant H191N, pH 8, 30°C Roseibium aggregatum
1.4.1.9 617
-
 NADH mutant P130V, pH 8, 30°C Roseibium aggregatum
1.4.1.9 688
-
 NADH mutant D153N/H191N, pH 8, 30°C Roseibium aggregatum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.4.1.9 8
-
-
 Roseibium aggregatum

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.4.1.9 8 10 more than 70% of maximum activity Roseibium aggregatum

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.4.1.9 6.7
-
 L-tert-leucine wild-type, pH 8, 30°C Roseibium aggregatum
1.4.1.9 43
-
 NADH wild-type, pH 8, 30°C Roseibium aggregatum
1.4.1.9 434.6
-
 NAD+ wild-type, pH 8, 30°C Roseibium aggregatum
1.4.1.9 808
-
 NADH wild-type, pH 8, 30°C Roseibium aggregatum
1.4.1.9 2058
-
 trimethylpyruvate mutant D153N, pH 8, 30°C Roseibium aggregatum
1.4.1.9 3618
-
 trimethylpyruvate mutant P130V, pH 8, 30°C Roseibium aggregatum
1.4.1.9 3620
-
 trimethylpyruvate wild-type, pH 8, 30°C Roseibium aggregatum
1.4.1.9 4794
-
 trimethylpyruvate mutant H191N, pH 8, 30°C Roseibium aggregatum
1.4.1.9 7165
-
 trimethylpyruvate mutant D153N/H191N, pH 8, 30°C Roseibium aggregatum
1.4.1.9 7936
-
 NADH mutant H191N, pH 8, 30°C Roseibium aggregatum
1.4.1.9 12348
-
 NADH mutant P130V, pH 8, 30°C Roseibium aggregatum
1.4.1.9 15013
-
 NADH mutant D153N, pH 8, 30°C Roseibium aggregatum
1.4.1.9 40465
-
 NADH mutant D153N/H191N, pH 8, 30°C Roseibium aggregatum