Cloned (Comment) | Organism |
---|---|
gene aldA, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
H449R | site-directed mutagenesis | Escherichia coli |
L158Y | site-directed mutagenesis | Escherichia coli |
additional information | calculated Gibbs free energy of protein-substrate interaction of enzyme mutants with D-glyceraldehyde as substrate | Escherichia coli |
N286A | site-directed mutagenesis | Escherichia coli |
N286C | site-directed mutagenesis | Escherichia coli |
N286D | site-directed mutagenesis | Escherichia coli |
N286E | site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme and is able to oxidate glyceraldehyde and methylglyoxal | Escherichia coli |
N286F | site-directed mutagenesis | Escherichia coli |
N286G | site-directed mutagenesis | Escherichia coli |
N286H | site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme and is able to oxidate glyceraldehyde and methylglyoxal, as well as acetaldehyde | Escherichia coli |
N286I | site-directed mutagenesis | Escherichia coli |
N286K | site-directed mutagenesis | Escherichia coli |
N286L | site-directed mutagenesis | Escherichia coli |
N286M | site-directed mutagenesis | Escherichia coli |
N286P | site-directed mutagenesis | Escherichia coli |
N286Q | site-directed mutagenesis | Escherichia coli |
N286R | site-directed mutagenesis | Escherichia coli |
N286S | site-directed mutagenesis | Escherichia coli |
N286T | site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme and is able to oxidate glyceraldehyde and methylglyoxal, as well as acetaldehyde | Escherichia coli |
N286V | site-directed mutagenesis | Escherichia coli |
N286W | site-directed mutagenesis | Escherichia coli |
N286Y | site-directed mutagenesis | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-lactaldehyde + NAD+ + H2O | Escherichia coli | - |
(S)-lactate + NADH + 2 H+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P25553 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by anion exchange chromatography to near homogeneity | Escherichia coli |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.74 | - |
purified recombinant enzyme, pH 7.0, 37°C, substrate (S)-lactaldehyde | Escherichia coli |
1.18 | - |
purified recombinant mutant N286H, pH 7.0, 37°C, substrate (S)-lactaldehyde | Escherichia coli |
1.78 | - |
purified recombinant mutant N286E, pH 7.0, 37°C, substrate (S)-lactaldehyde | Escherichia coli |
2.18 | - |
purified recombinant mutant N286T, pH 7.0, 37°C, substrate (S)-lactaldehyde | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-lactaldehyde + NAD+ + H2O | - |
Escherichia coli | (S)-lactate + NADH + 2 H+ | - |
? | |
additional information | residue Asn286 of L-lactaldehyde dehydrogenase plays an important structure role to substrate identification. The wild-type enzyme is not active with D-glyceraldehyde, methylglyoxal, acetaldehyde, L-lactate, or benzaldehyde | Escherichia coli | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 52200, recombinant enzyme, SDS-PAGE | Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
AldA | - |
Escherichia coli |
L-lactaldehyde dehydrogenase | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
15 | - |
recombinant mutant N286H | Escherichia coli |
40 | - |
recombinant wild-type enzyme | Escherichia coli |
45 | - |
recombinant mutants N286T and N286E | Escherichia coli |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | 55 | purified recombinant wild-type enzyme is stable at 40°C after incubation for 1 h, but its stability decreases rapidly above 40°C. The mutants N286E and N286H show the same stability, decreasing rapidly above 40°C and exhibiting 20% and 40% of their maximum activity at 45°C, respectively. In contrast, mutant N286T is stable above 50°C and retains 60% of its maximum activity at 55°C after incubation for 1 h | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Escherichia coli |
General Information | Comment | Organism |
---|---|---|
additional information | residue Asn286 of L-lactaldehyde dehydrogenase plays an important structure role to substrate identification, molecular structure modeling of wild-type enzyme and N286 mutants | Escherichia coli |