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Literature summary for 1.4.1.9 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
Search for more literature for 1.4.1.9

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Roseibium aggregatum

Protein Variants

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

KM Value [mM]

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

Organism

Organism UniProt Comment Textmining
Roseibium aggregatum
-
-
-

Specific Activity [micromol/min/mg]

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

Substrates and Products (Substrate)

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

Temperature Optimum [°C]

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

Temperature Stability [°C]

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

Turnover Number [1/s]

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

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
-
 Roseibium aggregatum

pH Range

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

kcat/KM [mM/s]

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