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Literature summary extracted from
- Thermostable alanine dehydrogenase from thermophilic Bacillus sphaericus DSM 462. Purification, characterization and kinetic mechanism (1990), Eur. J. Biochem., 191, 715-720.
General Stability
| EC Number | General Stability | Organism |
|---|---|---|
| 1.4.1.1 | SDS, 10 min, 37°C, retains activity with up to 0.03% SDS, loss of activity with 0.05% | Lysinibacillus sphaericus |
| 1.4.1.1 | stable to repeated freezing and thawing | Lysinibacillus sphaericus |
| 1.4.1.1 | Triton X-100, 10 min, 37°C, not inactivated by treatment with up to 4% | Lysinibacillus sphaericus |
Inhibitors
| EC Number | Inhibitors | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.4.1.1 | CuCl2 | - |
Lysinibacillus sphaericus |  |
| 1.4.1.1 | HgCl2 | - |
Lysinibacillus sphaericus | |
| 1.4.1.1 | NADH | product inhibition | Lysinibacillus sphaericus | |
| 1.4.1.1 | NH4+ | product inhibition | Lysinibacillus sphaericus | |
| 1.4.1.1 | PCMB | - |
Lysinibacillus sphaericus | |
| 1.4.1.1 | pyruvate | product inhibition | Lysinibacillus sphaericus | |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.4.1.1 | 0.1 | - |
NADH | - |
Lysinibacillus sphaericus | |
| 1.4.1.1 | 0.26 | - |
NAD+ | - |
Lysinibacillus sphaericus | |
| 1.4.1.1 | 0.5 | - |
pyruvate | - |
Lysinibacillus sphaericus | |
| 1.4.1.1 | 10.5 | - |
L-alanine | - |
Lysinibacillus sphaericus | |
| 1.4.1.1 | 38 | - |
NH4+ | - |
Lysinibacillus sphaericus | |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 1.4.1.1 | 38000 | - |
6 * 38000, SDS-PAGE | Lysinibacillus sphaericus |
| 1.4.1.1 | 230000 | - |
gel filtration | Lysinibacillus sphaericus |
Organic Solvent Stability
| EC Number | Organic Solvent | Comment | Organism |
|---|---|---|---|
| 1.4.1.1 | 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
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.4.1.1 | Lysinibacillus sphaericus | - |
- |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 1.4.1.1 | - |
Lysinibacillus sphaericus |
Reaction
| EC Number | Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|---|
| 1.4.1.1 | 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]
| EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|---|
| 1.4.1.1 | 13.4 | - |
- |
Lysinibacillus sphaericus |
Storage Stability
| EC Number | Storage Stability | Organism |
|---|---|---|
| 1.4.1.1 | -20°C, pH 7.2, more than 5 months | Lysinibacillus sphaericus |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.4.1.1 | 2-oxobutanoate + NH3 + NADH | 62% of the activity with pyruvate | Lysinibacillus sphaericus | 2-aminobutanoate + H2O + NAD+ | - |
? | |
| 1.4.1.1 | 3-fluoropyruvate + NH3 + NADH | 97% of the activity with pyruvate | Lysinibacillus sphaericus | 3-fluoro-L-alanine + H2O + NAD+ | - |
? | |
| 1.4.1.1 | 3-hydroxypyruvate + NH3 + NADH | 123% of the activity with pyruvate | Lysinibacillus sphaericus | L-Ser + H2O + NAD+ | - |
? | |
| 1.4.1.1 | L-2-aminobutanoate + H2O + NAD+ | 2.4% of the activity with L-Ala | Lysinibacillus sphaericus | 2-oxobutanoate + NH3 + NADH + H+ | - |
? | |
| 1.4.1.1 | 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 | |
| 1.4.1.1 | oxaloacetate + NH3 + NADH | 99% of the activity with pyruvate | Lysinibacillus sphaericus | L-Asp + H2O + NAD+ | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 1.4.1.1 | hexamer | 6 * 38000, SDS-PAGE | Lysinibacillus sphaericus |
Temperature Stability [°C]
| EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.4.1.1 | 37 | - |
pH 2.3-10.6, 10 min, stable | Lysinibacillus sphaericus |
| 1.4.1.1 | 75 | - |
60 min, pH 5.5-9.5, stable | Lysinibacillus sphaericus |
| 1.4.1.1 | 80 | - |
20 min, stable | Lysinibacillus sphaericus |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.4.1.1 | 8.2 | - |
amination of pyruvate | Lysinibacillus sphaericus |
pH Range
| EC Number | pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.4.1.1 | 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 |
| 1.4.1.1 | 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
| EC Number | pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.4.1.1 | 2.3 | 10.6 | 37°C, 10 min, stable | Lysinibacillus sphaericus |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.4.1.1 | NAD+ | - |
Lysinibacillus sphaericus | |
| 1.4.1.1 | NADH | - |
Lysinibacillus sphaericus | |
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