Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Levilactobacillus brevis |
Protein Variants | Comment | Organism |
---|---|---|
A90S | activity with NADPH as cofactor decreased to about 50% of wild-type, activity with NADH strongly decreased | Levilactobacillus brevis |
G37D/R38P | activity with NADH is decreased relative to Arg38Pro alone, but is higher than that of the wild-type enzyme | Levilactobacillus brevis |
M140I | no activity with NADPH as cofactor | Levilactobacillus brevis |
R38P | no activity with NADPH as cofactor, fourfold increase in activity with NADH | Levilactobacillus brevis |
V112D | no activity with NADPH as cofactor, strongly decreased activity with NADH | Levilactobacillus brevis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.017 | - |
NADPH | wild-type, pH 6.5, 30°C | Levilactobacillus brevis | |
0.028 | - |
NADPH | mutant R38P, pH 5.5, 30°C | Levilactobacillus brevis | |
0.12 | - |
NADH | mutant R38P, pH 5.5, 30°C | Levilactobacillus brevis | |
0.13 | - |
NADH | wild-type, pH 6.5, 30°C | Levilactobacillus brevis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Levilactobacillus brevis | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetophenone + NADH + H+ | - |
Levilactobacillus brevis | phenylethanol + NAD+ | - |
? | |
acetophenone + NADPH + H+ | - |
Levilactobacillus brevis | phenylethanol + NADP+ | - |
? |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
wild-type, half-life 160 h, mutant R38P, half-life 220 h | Levilactobacillus brevis |
42 | - |
wild-type, half-life 1.25 h, mutant R38P, half-life 3 h | Levilactobacillus brevis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
5.5 | - |
wild-type, cofactor NADH. Mutant R38P, both cofactors NADH and NADPH | Levilactobacillus brevis |
6 | 6.5 | wild-type, cofactor NADPH | Levilactobacillus brevis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADPH | clear preference for NADPH as cofactor. The phosphate group of NADPH interacts with both the side chains and main-chain amides of Arg38 and Thr15, stabilizing this cofactor significantly better than NADH | Levilactobacillus brevis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
75 | - |
NADH | wild-type, pH 6.5, 30°C | Levilactobacillus brevis | |
300 | - |
NADH | mutant R38P, pH 5.5, 30°C | Levilactobacillus brevis | |
5100 | - |
NADPH | mutant R38P, pH 5.5, 30°C | Levilactobacillus brevis | |
9300 | - |
NADPH | wild-type, pH 6.5, 30°C | Levilactobacillus brevis |