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Literature summary for 1.4.1.1 extracted from

  • Ohshima, T.; Sakane, M.; Yamazaki, T.; Soda, K.
    Thermostable alanine dehydrogenase from thermophilic Bacillus sphaericus DSM 462. Purification, characterization and kinetic mechanism (1990), Eur. J. Biochem., 191, 715-720.
    View publication on PubMed

General Stability

General Stability Organism
SDS, 10 min, 37°C, retains activity with up to 0.03% SDS, loss of activity with 0.05% Lysinibacillus sphaericus
stable to repeated freezing and thawing Lysinibacillus sphaericus
Triton X-100, 10 min, 37°C, not inactivated by treatment with up to 4% Lysinibacillus sphaericus

Inhibitors

Inhibitors Comment Organism Structure
CuCl2
-
Lysinibacillus sphaericus
HgCl2
-
Lysinibacillus sphaericus
NADH product inhibition Lysinibacillus sphaericus
NH4+ product inhibition Lysinibacillus sphaericus
PCMB
-
Lysinibacillus sphaericus
pyruvate product inhibition Lysinibacillus sphaericus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.1
-
NADH
-
Lysinibacillus sphaericus
0.26
-
NAD+
-
Lysinibacillus sphaericus
0.5
-
pyruvate
-
Lysinibacillus sphaericus
10.5
-
L-alanine
-
Lysinibacillus sphaericus
38
-
NH4+
-
Lysinibacillus sphaericus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
38000
-
6 * 38000, SDS-PAGE Lysinibacillus sphaericus
230000
-
gel filtration Lysinibacillus sphaericus

Organic Solvent Stability

Organic Solvent Comment Organism
Ethanol 10 min, 37°C, about 90% of activity remains after incubation with up to 35% ethanol, 50% activity with 48% ethanol Lysinibacillus sphaericus

Organism

Organism UniProt Comment Textmining
Lysinibacillus sphaericus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Lysinibacillus sphaericus

Reaction

Reaction Comment Organism Reaction ID
L-alanine + H2O + NAD+ = pyruvate + NH3 + NADH + H+ reductive amination proceeds through a sequential mechanism containing partially random binding. NADH binds first to the enzyme, and then pyruvate and ammonia bind in a random fashion Lysinibacillus sphaericus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
13.4
-
-
Lysinibacillus sphaericus

Storage Stability

Storage Stability Organism
-20°C, pH 7.2, more than 5 months Lysinibacillus sphaericus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-oxobutanoate + NH3 + NADH 62% of the activity with pyruvate Lysinibacillus sphaericus 2-aminobutanoate + H2O + NAD+
-
?
3-fluoropyruvate + NH3 + NADH 97% of the activity with pyruvate Lysinibacillus sphaericus 3-fluoro-L-alanine + H2O + NAD+
-
?
3-hydroxypyruvate + NH3 + NADH 123% of the activity with pyruvate Lysinibacillus sphaericus L-Ser + H2O + NAD+
-
?
L-2-aminobutanoate + H2O + NAD+ 2.4% of the activity with L-Ala Lysinibacillus sphaericus 2-oxobutanoate + NH3 + NADH + H+
-
?
L-Ala + H2O + NAD+ the reaction rate for deamination of pyruvate at the optimum is about 2.8times higher than that for amination of L-Ala at the optimum Lysinibacillus sphaericus pyruvate + NH3 + NADH
-
r
oxaloacetate + NH3 + NADH 99% of the activity with pyruvate Lysinibacillus sphaericus L-Asp + H2O + NAD+
-
?

Subunits

Subunits Comment Organism
hexamer 6 * 38000, SDS-PAGE Lysinibacillus sphaericus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
37
-
pH 2.3-10.6, 10 min, stable Lysinibacillus sphaericus
75
-
60 min, pH 5.5-9.5, stable Lysinibacillus sphaericus
80
-
20 min, stable Lysinibacillus sphaericus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.2
-
amination of pyruvate Lysinibacillus sphaericus

pH Range

pH Minimum pH Maximum Comment Organism
7 10 pH 7.0: about 50% of maximal activity, pH 10.0: about 50% of maximal activity, oxidative deamination of L-Ala Lysinibacillus sphaericus
8 11 pH 8.0: about 50% of maximal activity, pH 11.0: about 95% of maximal activity, reductive amination of pyruvate Lysinibacillus sphaericus

pH Stability

pH Stability pH Stability Maximum Comment Organism
2.3 10.6 37°C, 10 min, stable Lysinibacillus sphaericus

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Lysinibacillus sphaericus
NADH
-
Lysinibacillus sphaericus