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Literature summary for 1.4.1.1 extracted from

  • Ye, W.; Huo, G.; Chen, J.; Liu, F.; Yin, J.; Yang, L.; Ma, X.
    Heterologous expression of the Bacillus subtilis (natto) alanine dehydrogenase in Escherichia coli and Lactococcus lactis (2009), Microbiol. Res., 165, 268-775.
    View publication on PubMed

Application

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(Commentary)

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 [Da] Molecular Weight Maximum [Da] Comment Organism
44000
-
AlaDH, SDS-PAGE Bacillus subtilis
70000
-
GST-AlaDH fusion protein, SDS-PAGE Bacillus subtilis

Organism

Organism UniProt Comment Textmining
Bacillus subtilis Q08352
-
-

Purification (Commentary)

Purification (Comment) Organism
the GST-AlaDH fusion protein is released by cleaving with PreScission protease Bacillus subtilis

Specific Activity [micromol/min/mg]

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 and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
pyruvate + NH3 + NADH
-
Bacillus subtilis L-alanine + H2O + NAD+
-
?

Synonyms

Synonyms Comment Organism
ALADH
-
Bacillus subtilis
alanine dehydrogenase
-
Bacillus subtilis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Bacillus subtilis
42
-
assay at Bacillus subtilis

Cofactor

Cofactor Comment Organism Structure
NADH
-
Bacillus subtilis