Application | Comment | Organism |
---|---|---|
biotechnology | heterologous expression of the Bacillus subtilis AlaDH in Lactococcus lactis using the promoter of lactate dehydrogenase from Streptococcus thermophilus leads to a better alanine production in the recombinant strain | Bacillus subtilis |
Cloned (Comment) | Organism |
---|---|
AlaDH gene is cloned into a pGEX6p-1 and expressed in Escherichia coli JM109. AlaDH gene is then cloned intoa vector pNZ8148 to generate a vector pNZ8148/ald. The same AlaDH gene is placed downstream of the lactate dehydrogenase (ldh) promoter from Streptococcus thermophilus to generate pNZ273/ldhp/AlaDH. The pNZ8148/AlaDH and pNZ273/ldhp/AlaDH are introduced separately in Lactococcus lactis NZ9000. As a result of over-expressed AlaDH, the production of alanine detected by HPLC in Lactococcus lactis NZ9000 carrying pNZ273/ldhp/ald reaches 52 mg/ml, an approximately 26fold increase compared to the parent strain Lactococcus lactis NZ9000, but not in Lactococcus lactis NZ9000 carrying pNZ8148/ALaDH | Bacillus subtilis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
44000 | - |
AlaDH, SDS-PAGE | Bacillus subtilis |
70000 | - |
GST-AlaDH fusion protein, SDS-PAGE | Bacillus subtilis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus subtilis | Q08352 | - |
- |
Purification (Comment) | Organism |
---|---|
the GST-AlaDH fusion protein is released by cleaving with PreScission protease | Bacillus subtilis |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
45.2 | - |
enzymatic activity of AlaDH expressed in Escherichia coli JM109 at 42.1°C | Bacillus subtilis |
48.3 | - |
enzymatic activity of AlaDH expressed in Escherichia coli JM109 at 30.1°C | Bacillus subtilis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
pyruvate + NH3 + NADH | - |
Bacillus subtilis | L-alanine + H2O + NAD+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
ALADH | - |
Bacillus subtilis |
alanine dehydrogenase | - |
Bacillus subtilis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Bacillus subtilis |
42 | - |
assay at | Bacillus subtilis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | - |
Bacillus subtilis |