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Literature summary for 1.4.1.9 extracted from

  • Luo, W.; Zhu, J.; Zhao, Y.; Zhang, H.; Yang, X.; Liu, Y.; Rao, Z.; Yu, X.
    Cloning and expression of a novel leucine dehydrogenase characterization and L-tert-leucine production (2020), Front. Bioeng. Biotechnol., 8, 186 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.4.1.9

Activating Compound

Activating Compound Comment Organism Structure
additional information not activating nor inhibitory: Na+, Ca2+, Mg2+, Mn2+, Cu2+, and Zn2+ Exiguobacterium sibiricum

Application

Application Comment Organism
synthesis enzyme coupled with recombinant formate dehydrogenase is used to catalyze trimethylpyruvic acid through reductive amination to generate enantiopure L-tert-leucine. Using a fed-batch feeding strategy, up to 0.8 M of trimethylpyruvate is transformed to L-tert-leucine, with an average conversion rate of 81% and L-tert-leucine concentration of 65.6 g/l Exiguobacterium sibiricum

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Exiguobacterium sibiricum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.624
-
 NADH pH 8.5, 30°C Exiguobacterium sibiricum
0.88
-
 L-leucine pH 8.5, 30°C Exiguobacterium sibiricum
1.46
-
 NAD+ pH 8.5, 30°C Exiguobacterium sibiricum
5.96
-
 trimethylpyruvate pH 8.5, 30°C Exiguobacterium sibiricum

Metals/Ions

Metals/Ions Comment Organism Structure
EDTA 1 mM, 86% residual activity Exiguobacterium sibiricum
Fe2+ 1 mM, 60% residual activity Exiguobacterium sibiricum
Fe3+ 1 mM, 48% residual activity Exiguobacterium sibiricum
additional information not activating nor inhibitory: Na+, Ca2+, Mg2+, Mn2+, Cu2+, and Zn2+ Exiguobacterium sibiricum

Organism

Organism UniProt Comment Textmining
Exiguobacterium sibiricum B1YLR3
-
-
Exiguobacterium sibiricum DSM 17290 B1YLR3
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-oxobutanoate + NH3 + NADH + H+
-
 Exiguobacterium sibiricum L-2-aminobutanoate + H2O + NAD+
-
 ?
2-oxobutanoate + NH3 + NADH + H+
-
 Exiguobacterium sibiricum DSM 17290 L-2-aminobutanoate + H2O + NAD+
-
 ?
4-methyl-2-oxopentanoate + NH3 + NADH + H+
-
 Exiguobacterium sibiricum L-leucine + H2O + NAD+
-
 ?
4-methyl-2-oxopentanoate + NH3 + NADH + H+
-
 Exiguobacterium sibiricum DSM 17290 L-leucine + H2O + NAD+
-
 ?
L-isoleucine + NH3 + NADH + H+
-
 Exiguobacterium sibiricum 3-methyl-2-oxopentanoate + H2O + NAD+
-
 ?
L-leucine + H2O + NAD+
-
 Exiguobacterium sibiricum 4-methyl-2-oxopentanoate + NH3 + NADH + H+
-
 ?
L-leucine + H2O + NAD+
-
 Exiguobacterium sibiricum DSM 17290 4-methyl-2-oxopentanoate + NH3 + NADH + H+
-
 ?
L-valine + NH3 + NADH + H+
-
 Exiguobacterium sibiricum 2-hydroxy-3-methylbutanoate + H2O + NAD+
-
 ?
phenylglyoxylate + NH3 + NADH + H+
-
 Exiguobacterium sibiricum amino(phenyl)acetate + H2O + NAD+
-
 ?
pyruvate + NH3 + NADH + H+
-
 Exiguobacterium sibiricum L-alanine + H2O + NAD+
-
 ?
pyruvate + NH3 + NADH + H+
-
 Exiguobacterium sibiricum DSM 17290 L-alanine + H2O + NAD+
-
 ?
trimethylpyruvate + NH3 + NADH + H+
-
 Exiguobacterium sibiricum L-tert-leucine + H2O + NAD+
-
 ?
trimethylpyruvate + NH3 + NADH + H+
-
 Exiguobacterium sibiricum DSM 17290 L-tert-leucine + H2O + NAD+
-
 ?

Subunits

Subunits Comment Organism
? x * 40000, SDS-PAGE and calculated from sequence Exiguobacterium sibiricum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
40
-
-
 Exiguobacterium sibiricum

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
50
-
 half-life 14.7 h Exiguobacterium sibiricum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.3
-
 L-leucine pH 8.5, 30°C Exiguobacterium sibiricum
8.4
-
 NAD+ pH 8.5, 30°C Exiguobacterium sibiricum
45.4
-
 trimethylpyruvate pH 8.5, 30°C Exiguobacterium sibiricum
76.9
-
 NADH pH 8.5, 30°C Exiguobacterium sibiricum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9
-
-
 Exiguobacterium sibiricum

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
4.9
-
 L-leucine pH 8.5, 30°C Exiguobacterium sibiricum
5.8
-
 NAD+ pH 8.5, 30°C Exiguobacterium sibiricum
7.6
-
 trimethylpyruvate pH 8.5, 30°C Exiguobacterium sibiricum
123.2
-
 NADH pH 8.5, 30°C Exiguobacterium sibiricum