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Literature summary for 1.4.1.9 extracted from

  • Zhou, F.; Mu, X.; Nie, Y.; Xu, Y.
    Enhanced catalytic efficiency and coenzyme affinity of leucine dehydrogenase by comprehensive screening strategy for L-tert-leucine synthesis (2021), Appl. Microbiol. Biotechnol., 105, 3625-3634 .
    View publication on PubMed
Search for more literature for 1.4.1.9

Application

Application Comment Organism
synthesis production of L-tert-leucine. A coupled reaction comprising LeuDH with glucose dehydrogenase of Bacillus amyloliquefaciens results in substrate inhibition at high trimethylpyruvate concentrations (0.5 M), which is overcome by batch-feeding of the substrate. The total turnover number and specific space-time conversion of 0.57 M substrate increases to 11400 and 22.8 mmol per h and l and g, respectively Bacillus cereus

Cloned(Commentary)

Cloned (Comment) Organism
-
 Bacillus cereus

Protein Variants

Protein Variants Comment Organism
D126E 2fold increase in kcat/Km value for trimethylpyruvate Bacillus cereus
E116V 3fold increase in kcat/Km value for trimethylpyruvate Bacillus cereus
E24A/D126E affinity for trimethylpyruvate and NADH is increased by 7.7- and 2.8fold, respectively Bacillus cereus
E24V/E116V 5fold increase in kcat/Km value for trimethylpyruvate Bacillus cereus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.1
-
 NADH mutant E24V/E116V, pH 8.5, 30°C Bacillus cereus
0.154
-
 NADH mutant E24A/D126E, pH 8.5, 30°C Bacillus cereus
0.25
-
 NADH mutant E116V, pH 8.5, 30°C Bacillus cereus
0.27
-
 NADH mutant D126E, pH 8.5, 30°C Bacillus cereus
0.28
-
 NADH wild-type, pH 8.5, 30°C Bacillus cereus
7.9
-
 trimethylpyruvate mutant E24V/E116V, pH 8.5, 30°C Bacillus cereus
10.7
-
 trimethylpyruvate mutant E116V, pH 8.5, 30°C Bacillus cereus
17.6
-
 trimethylpyruvate mutant E24A/D126E, pH 8.5, 30°C Bacillus cereus
23.2
-
 trimethylpyruvate mutant D126E, pH 8.5, 30°C Bacillus cereus
60.8
-
 trimethylpyruvate wild-type, pH 8.5, 30°C Bacillus cereus

Organism

Organism UniProt Comment Textmining
Bacillus cereus A0A068FCF1
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
trimethylpyruvate + NH3 + NADH + H+
-
 Bacillus cereus L-tert-leucine + H2O + NAD+
-
 ?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
33.7
-
 NADH wild-type, pH 8.5, 30°C Bacillus cereus
35.1
-
 NADH mutant E24V/E116V, pH 8.5, 30°C Bacillus cereus
42.6
-
 NADH mutant E24A/D126E, pH 8.5, 30°C Bacillus cereus
50.9
-
 NADH mutant D126E, pH 8.5, 30°C Bacillus cereus
60.5
-
 NADH mutant E116V, pH 8.5, 30°C Bacillus cereus
60.5
-
 trimethylpyruvate mutant E116V, pH 8.5, 30°C Bacillus cereus
66.2
-
 trimethylpyruvate mutant E24V/E116V, pH 8.5, 30°C Bacillus cereus
77.4
-
 trimethylpyruvate mutant D126E, pH 8.5, 30°C Bacillus cereus
93.3
-
 trimethylpyruvate mutant E24A/D126E, pH 8.5, 30°C Bacillus cereus
94.1
-
 trimethylpyruvate wild-type, pH 8.5, 30°C Bacillus cereus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.55
-
 trimethylpyruvate wild-type, pH 8.5, 30°C Bacillus cereus
3.34
-
 trimethylpyruvate mutant D126E, pH 8.5, 30°C Bacillus cereus
5.31
-
 trimethylpyruvate mutant E24A/D126E, pH 8.5, 30°C Bacillus cereus
5.63
-
 trimethylpyruvate mutant E116V, pH 8.5, 30°C Bacillus cereus
8.36
-
 trimethylpyruvate mutant E24V/E116V, pH 8.5, 30°C Bacillus cereus
118.5
-
 NADH wild-type, pH 8.5, 30°C Bacillus cereus
188.6
-
 NADH mutant D126E, pH 8.5, 30°C Bacillus cereus
241.9
-
 NADH mutant E116V, pH 8.5, 30°C Bacillus cereus
276.8
-
 NADH mutant E24A/D126E, pH 8.5, 30°C Bacillus cereus
350.9
-
 NADH mutant E24V/E116V, pH 8.5, 30°C Bacillus cereus