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Literature summary 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

Activating Compound

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

Application

EC Number Application Comment Organism
1.4.1.9 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)

EC Number Cloned (Comment) Organism
1.4.1.9 expression in Escherichia coli Exiguobacterium sibiricum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.4.1.9 0.624
-
 NADH pH 8.5, 30°C Exiguobacterium sibiricum
1.4.1.9 0.88
-
 L-leucine pH 8.5, 30°C Exiguobacterium sibiricum
1.4.1.9 1.46
-
 NAD+ pH 8.5, 30°C Exiguobacterium sibiricum
1.4.1.9 5.96
-
 trimethylpyruvate pH 8.5, 30°C Exiguobacterium sibiricum

Metals/Ions

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

Organism

EC Number Organism UniProt Comment Textmining
1.4.1.9 Exiguobacterium sibiricum B1YLR3
-
-
1.4.1.9 Exiguobacterium sibiricum DSM 17290 B1YLR3
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.4.1.9 2-oxobutanoate + NH3 + NADH + H+
-
 Exiguobacterium sibiricum L-2-aminobutanoate + H2O + NAD+
-
 ?
1.4.1.9 2-oxobutanoate + NH3 + NADH + H+
-
 Exiguobacterium sibiricum DSM 17290 L-2-aminobutanoate + H2O + NAD+
-
 ?
1.4.1.9 4-methyl-2-oxopentanoate + NH3 + NADH + H+
-
 Exiguobacterium sibiricum L-leucine + H2O + NAD+
-
 ?
1.4.1.9 4-methyl-2-oxopentanoate + NH3 + NADH + H+
-
 Exiguobacterium sibiricum DSM 17290 L-leucine + H2O + NAD+
-
 ?
1.4.1.9 L-isoleucine + NH3 + NADH + H+
-
 Exiguobacterium sibiricum 3-methyl-2-oxopentanoate + H2O + NAD+
-
 ?
1.4.1.9 L-leucine + H2O + NAD+
-
 Exiguobacterium sibiricum 4-methyl-2-oxopentanoate + NH3 + NADH + H+
-
 ?
1.4.1.9 L-leucine + H2O + NAD+
-
 Exiguobacterium sibiricum DSM 17290 4-methyl-2-oxopentanoate + NH3 + NADH + H+
-
 ?
1.4.1.9 L-valine + NH3 + NADH + H+
-
 Exiguobacterium sibiricum 2-hydroxy-3-methylbutanoate + H2O + NAD+
-
 ?
1.4.1.9 phenylglyoxylate + NH3 + NADH + H+
-
 Exiguobacterium sibiricum amino(phenyl)acetate + H2O + NAD+
-
 ?
1.4.1.9 pyruvate + NH3 + NADH + H+
-
 Exiguobacterium sibiricum L-alanine + H2O + NAD+
-
 ?
1.4.1.9 pyruvate + NH3 + NADH + H+
-
 Exiguobacterium sibiricum DSM 17290 L-alanine + H2O + NAD+
-
 ?
1.4.1.9 trimethylpyruvate + NH3 + NADH + H+
-
 Exiguobacterium sibiricum L-tert-leucine + H2O + NAD+
-
 ?
1.4.1.9 trimethylpyruvate + NH3 + NADH + H+
-
 Exiguobacterium sibiricum DSM 17290 L-tert-leucine + H2O + NAD+
-
 ?

Subunits

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

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.4.1.9 40
-
-
 Exiguobacterium sibiricum

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.4.1.9 50
-
 half-life 14.7 h Exiguobacterium sibiricum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.4.1.9 4.3
-
 L-leucine pH 8.5, 30°C Exiguobacterium sibiricum
1.4.1.9 8.4
-
 NAD+ pH 8.5, 30°C Exiguobacterium sibiricum
1.4.1.9 45.4
-
 trimethylpyruvate pH 8.5, 30°C Exiguobacterium sibiricum
1.4.1.9 76.9
-
 NADH pH 8.5, 30°C Exiguobacterium sibiricum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.4.1.9 9
-
-
 Exiguobacterium sibiricum

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 4.9
-
 L-leucine pH 8.5, 30°C Exiguobacterium sibiricum
1.4.1.9 5.8
-
 NAD+ pH 8.5, 30°C Exiguobacterium sibiricum
1.4.1.9 7.6
-
 trimethylpyruvate pH 8.5, 30°C Exiguobacterium sibiricum
1.4.1.9 123.2
-
 NADH pH 8.5, 30°C Exiguobacterium sibiricum