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Literature summary for 1.1.1.103 extracted from

  • Nakano, S.; Okazaki, S.; Tokiwa, H.; Asano, Y.
    Binding of NAD+ and L-threonine induces stepwise structural and flexibility changes in Cupriavidus necator L-threonine dehydrogenase (2014), J. Biol. Chem., 289, 10445-10454.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Cupriavidus necator

Crystallization (Commentary)

Crystallization (Comment) Organism
hanging drop vapor diffusion method, using 20% (w/v) PEG 3350 and 0.2 M CaCl2 Cupriavidus necator

Protein Variants

Protein Variants Comment Organism
G184A the catalytic efficiency of the mutant is 330fold lower than that of the wild type enzyme Cupriavidus necator
L80G the catalytic efficiency of the mutant is 3300fold lower than that of the wild type enzyme Cupriavidus necator
T186N the catalytic efficiency of the mutant is 330fold lower than that of the wild type enzyme Cupriavidus necator

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.097
-
NAD+ mutant enzyme T186N, at pH 10.0 and 30°C Cupriavidus necator
0.115
-
NAD+ mutant enzyme L80G, at pH 10.0 and 30°C Cupriavidus necator
0.12
-
NAD+ wild type enzyme, at pH 10.0 and 30°C Cupriavidus necator
0.156
-
NAD+ mutant enzyme G184A, at pH 10.0 and 30°C Cupriavidus necator
15.2
-
L-threonine wild type enzyme, at pH 10.0 and 30°C Cupriavidus necator

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-threonine + NAD+ Cupriavidus necator
-
(2S)-2-amino-3-oxobutanoate + NADH + H+
-
?

Organism

Organism UniProt Comment Textmining
Cupriavidus necator
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni2+-Sepharose column chromatography and Superdex 75 gel filtration Cupriavidus necator

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-threonine + NAD+
-
Cupriavidus necator (2S)-2-amino-3-oxobutanoate + NADH + H+
-
?

Synonyms

Synonyms Comment Organism
L-ThrDH
-
Cupriavidus necator
L-threonine dehydrogenase
-
Cupriavidus necator
ThrDH
-
Cupriavidus necator

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
50
-
L-threonine wild type enzyme, at pH 10.0 and 30°C Cupriavidus necator

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Cupriavidus necator

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.001
-
NAD+ mutant enzyme L80G, at pH 10.0 and 30°C Cupriavidus necator
0.01
-
NAD+ mutant enzyme G184A, at pH 10.0 and 30°C Cupriavidus necator
0.01
-
NAD+ mutant enzyme T186N, at pH 10.0 and 30°C Cupriavidus necator
3.3
-
L-threonine wild type enzyme, at pH 10.0 and 30°C Cupriavidus necator