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Literature summary for 1.4.1.1 extracted from

  • Sakamoto, Y.; Nagata, S.; Esaki, N.; Tanaka, H.; Soda, K.
    Gene cloning, purification and characterization of thermostable alanine dehydrogenase of bacillus stearothermophilus (1990), J. Ferment. Bioeng., 69, 154-158.
No PubMed abstract available

Cloned(Commentary)

Cloned (Comment) Organism
cloned and expressed in Escherichia coli Geobacillus stearothermophilus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.077
-
NADH
-
Geobacillus stearothermophilus
1.67
-
NAD+
-
Geobacillus stearothermophilus
5
-
pyruvate
-
Geobacillus stearothermophilus
13.3
-
L-alanine
-
Geobacillus stearothermophilus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
40000
-
6 * 40000, recombinant enzyme, SDS-PAGE Geobacillus stearothermophilus
240000
-
recombinant enzyme, equilibrium sedimentation, gel filtration Geobacillus stearothermophilus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-Ala + H2O + NAD+ Geobacillus stearothermophilus
-
pyruvate + NH3 + NADH
-
?

Organism

Organism UniProt Comment Textmining
Geobacillus stearothermophilus
-
gene cloned and expressed in Escherichia coli
-

Purification (Commentary)

Purification (Comment) Organism
gene cloned and expressed in Escherichia coli Geobacillus stearothermophilus

Reaction

Reaction Comment Organism Reaction ID
L-alanine + H2O + NAD+ = pyruvate + NH3 + NADH + H+ reductive amination: sequential mechanism with partially random binding Geobacillus stearothermophilus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
75
-
-
Geobacillus stearothermophilus

Storage Stability

Storage Stability Organism
4°C, 0.15 M KCl, stable for more than 2 months Geobacillus stearothermophilus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-oxobutanoate + NH3 + NADH
-
Geobacillus stearothermophilus 2-aminobutanoate + H2O + NAD+
-
r
L-2-aminobutanoate + H2O + NAD+ 7.5% of the activity with L-Ala Geobacillus stearothermophilus 2-oxobutanoate + NH3 + NADH + H+
-
r
L-2-aminobutanoate + H2O + NAD+ in the reverse reaction 2-oxobutanoate reacts with 79% of the activity with pyruvate Geobacillus stearothermophilus 2-oxobutanoate + NH3 + NADH + H+
-
r
L-Ala + H2O + NAD+
-
Geobacillus stearothermophilus pyruvate + NH3 + NADH
-
?
L-Ala + H2O + NAD+
-
Geobacillus stearothermophilus pyruvate + NH3 + NADH
-
r
L-norvaline + H2O + NAD+ 1.5% of the activity with L-Ala Geobacillus stearothermophilus 2-oxopentanoate + NH3 + NADH
-
r
L-norvaline + H2O + NAD+ in the reverse reaction 2-oxopentanoate reacts with 6.6% of the activity of pyruvate Geobacillus stearothermophilus 2-oxopentanoate + NH3 + NADH
-
r
L-norvaline + H2O + NAD+ in the reverse reaction 2-oxopentanoate reacts with 0.12% of the activity of pyruvate Geobacillus stearothermophilus 2-oxopentanoate + NH3 + NADH
-
r
L-Ser + H2O + NAD+ 3.5% of the activity with L-Ala Geobacillus stearothermophilus 3-hydroxypyruvate + NH3 + NADH
-
r
L-Ser + H2O + NAD+ in the reverse reaction 3-hydroxypyruvate reacts with 0.4% of the activity with pyruvate Geobacillus stearothermophilus 3-hydroxypyruvate + NH3 + NADH
-
r

Subunits

Subunits Comment Organism
hexamer 6 * 40000, recombinant enzyme, SDS-PAGE Geobacillus stearothermophilus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
55
-
pH 6.0-11.0, 10 min, stable Geobacillus stearothermophilus
75
-
30 min, stable Geobacillus stearothermophilus
85
-
5 min, substantial loss of activity Geobacillus stearothermophilus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9.5
-
reductive amination of pyruvate Geobacillus stearothermophilus
10.7 11.2 oxidative deamination Geobacillus stearothermophilus

pH Range

pH Minimum pH Maximum Comment Organism
7.8 10.2 pH 7.8: 54% of activity maximum, pH 10.2: 72% of activity maximum, reductive amination of pyruvate Geobacillus stearothermophilus
9.3 11.8 pH 9.3: about 55% of activity maximum, pH 11.8: about 63% of activity maximum, oxidative deamination Geobacillus stearothermophilus

pH Stability

pH Stability pH Stability Maximum Comment Organism
6 10 55°C, 10 min, stable Geobacillus stearothermophilus

Cofactor

Cofactor Comment Organism Structure
NAD+ specific for Geobacillus stearothermophilus
NADH specific for Geobacillus stearothermophilus