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Literature summary extracted from
- 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)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 1.4.1.1 | cloned and expressed in Escherichia coli | Geobacillus stearothermophilus |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.4.1.1 | 0.077 | - |
NADH | - |
Geobacillus stearothermophilus |  |
| 1.4.1.1 | 1.67 | - |
NAD+ | - |
Geobacillus stearothermophilus | |
| 1.4.1.1 | 5 | - |
pyruvate | - |
Geobacillus stearothermophilus | |
| 1.4.1.1 | 13.3 | - |
L-alanine | - |
Geobacillus stearothermophilus | |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 1.4.1.1 | 40000 | - |
6 * 40000, recombinant enzyme, SDS-PAGE | Geobacillus stearothermophilus |
| 1.4.1.1 | 240000 | - |
recombinant enzyme, equilibrium sedimentation, gel filtration | Geobacillus stearothermophilus |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.4.1.1 | L-Ala + H2O + NAD+ | Geobacillus stearothermophilus | - |
pyruvate + NH3 + NADH | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.4.1.1 | Geobacillus stearothermophilus | - |
gene cloned and expressed in Escherichia coli | - |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 1.4.1.1 | gene cloned and expressed in Escherichia coli | Geobacillus stearothermophilus |
Reaction
| EC Number | Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|---|
| 1.4.1.1 | L-alanine + H2O + NAD+ = pyruvate + NH3 + NADH + H+ | reductive amination: sequential mechanism with partially random binding | Geobacillus stearothermophilus |
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 | 75 | - |
- |
Geobacillus stearothermophilus |
Storage Stability
| EC Number | Storage Stability | Organism |
|---|---|---|
| 1.4.1.1 | 4°C, 0.15 M KCl, stable for more than 2 months | Geobacillus stearothermophilus |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.4.1.1 | 2-oxobutanoate + NH3 + NADH | - |
Geobacillus stearothermophilus | 2-aminobutanoate + H2O + NAD+ | - |
r | |
| 1.4.1.1 | L-2-aminobutanoate + H2O + NAD+ | 7.5% of the activity with L-Ala | Geobacillus stearothermophilus | 2-oxobutanoate + NH3 + NADH + H+ | - |
r | |
| 1.4.1.1 | 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 | |
| 1.4.1.1 | L-Ala + H2O + NAD+ | - |
Geobacillus stearothermophilus | pyruvate + NH3 + NADH | - |
? | |
| 1.4.1.1 | L-Ala + H2O + NAD+ | - |
Geobacillus stearothermophilus | pyruvate + NH3 + NADH | - |
r | |
| 1.4.1.1 | L-norvaline + H2O + NAD+ | 1.5% of the activity with L-Ala | Geobacillus stearothermophilus | 2-oxopentanoate + NH3 + NADH | - |
r | |
| 1.4.1.1 | 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 | |
| 1.4.1.1 | 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 | |
| 1.4.1.1 | L-Ser + H2O + NAD+ | 3.5% of the activity with L-Ala | Geobacillus stearothermophilus | 3-hydroxypyruvate + NH3 + NADH | - |
r | |
| 1.4.1.1 | 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
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 1.4.1.1 | hexamer | 6 * 40000, recombinant enzyme, SDS-PAGE | Geobacillus stearothermophilus |
Temperature Stability [°C]
| EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.4.1.1 | 55 | - |
pH 6.0-11.0, 10 min, stable | Geobacillus stearothermophilus |
| 1.4.1.1 | 75 | - |
30 min, stable | Geobacillus stearothermophilus |
| 1.4.1.1 | 85 | - |
5 min, substantial loss of activity | Geobacillus stearothermophilus |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.4.1.1 | 9.5 | - |
reductive amination of pyruvate | Geobacillus stearothermophilus |
| 1.4.1.1 | 10.7 | 11.2 | oxidative deamination | Geobacillus stearothermophilus |
pH Range
| EC Number | pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.4.1.1 | 7.8 | 10.2 | pH 7.8: 54% of activity maximum, pH 10.2: 72% of activity maximum, reductive amination of pyruvate | Geobacillus stearothermophilus |
| 1.4.1.1 | 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
| EC Number | pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.4.1.1 | 6 | 10 | 55°C, 10 min, stable | Geobacillus stearothermophilus |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.4.1.1 | NAD+ | specific for | Geobacillus stearothermophilus | |
| 1.4.1.1 | NADH | specific for | Geobacillus stearothermophilus | |
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