Cloned (Comment) | Organism |
---|---|
cloned into the pkk177-3H vector, transformation in Escherichia coli | Levilactobacillus brevis |
Crystallization (Comment) | Organism |
---|---|
16°C, wild-type RADH: 15 mg/ml stock solution (21% polyethyleneglycol monomethyl ether 550, 0.1 M Tris-HCl, 50 nM MgCl2, 40 mM acetophenone, 10 mM NADP+), growth for 3 weeks, max. size 0.25 mm x 0.25 mm x 1.0 mm, RADH-G37D: 21 mg/ml stock solution (methyl-2,4-pentanediol, 20% polyethyleneglycol 400 or polyethyleneglycol monomethyl ether 550, 0.1 M Hepes, 50 mM MgCl2 59 mM 1-phenylethanol, 25 mM NADH), growth 1 week, max. size 0.4 mm x 0.5 mm x 1.7 mm | Levilactobacillus brevis |
Protein Variants | Comment | Organism |
---|---|---|
G37D | regular PCR with site-specific mutation for both DNA strands, the gene is divided into two separate amplification products, preference for NAD+ rather than NADP+, depends on an a interaction between the adenosine ribose moiety of NAD+ and the inserted aspartate side chain | Levilactobacillus brevis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.015 | - |
NADP+ | reduction, recombinant wild-type, 30°C, pH 7.0 | Levilactobacillus brevis | |
0.04 | - |
NADPH | oxidation, recombinante wild-typ, 30°C, pH 7.0 | Levilactobacillus brevis | |
0.75 | - |
NAD+ | reduction, recombinant mutant G37D, 30°C, pH 7.0 | Levilactobacillus brevis | |
1.6 | - |
NADH | oxidation, recombinante mutant G37D, 30°C, pH 7.0 | Levilactobacillus brevis | |
2.8 | - |
acetophenone | cofactor NADPH, reduction, recombinante wild-type, 30°C, pH 7.0 | Levilactobacillus brevis | |
2.9 | - |
1-phenylethanol | cofactor NADP+, oxidation, recombinate wild-type, 30°C, pH 7.0 | Levilactobacillus brevis | |
9.1 | - |
NADP+ | reduction, recombinant mutant G37D, 30°C, pH 7.0 | Levilactobacillus brevis | |
10.8 | - |
1-phenylethanol | NAD+, oxidation, recombinante mutant G37D, 30°C, pH 7.0 | Levilactobacillus brevis | |
10.9 | - |
acetophenone | cofactor NADH, reduction, recombinante mutant G37D, 30°C, pH 7.0 | Levilactobacillus brevis | |
55.5 | - |
1-phenylethanol | cofactor NADP+, oxidation, recombinate mutant G37D, 30°C, pH 7.0 | Levilactobacillus brevis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Levilactobacillus brevis | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetophenone + NADH + H+ | recombinant mutant G37D | Levilactobacillus brevis | 1-phenylethanol + NAD+ | recombinate mutant G37D | r | |
acetophenone + NADPH + H+ | recombinant wild-type | Levilactobacillus brevis | 1-phenylethanol + NADP+ | recombinant wild-type | r | |
phenylethanol + NADP+ | - |
Levilactobacillus brevis | phenylacetaldehyde + NADPH + H+ | - |
r |
Synonyms | Comment | Organism |
---|---|---|
RADH | - |
Levilactobacillus brevis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
- |
Levilactobacillus brevis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
4.4 | - |
NADP+ | reduction, recombinant wild-type, 30°C, pH 7.0 | Levilactobacillus brevis | |
5.4 | - |
Phenylethanol | oxidation, cosubstrate NADP+, recombinant wild-type, 30°C, pH 7.0 | Levilactobacillus brevis | |
5.9 | - |
NADP+ | reduction, recombinant mutant G37D, 30°C, pH 7.0 | Levilactobacillus brevis | |
8.2 | - |
Phenylethanol | oxidation, cosubstrate NADP+, recombinant mutant G37D, 30°C, pH 7.0 | Levilactobacillus brevis | |
9.5 | - |
Phenylethanol | oxidation, cosubstrate NAD+, recombinant mutant G37D, 30°C, pH 7.0 | Levilactobacillus brevis | |
9.9 | - |
NAD+ | reduction, recombinant mutant G37D, 30°C, pH 7.0 | Levilactobacillus brevis | |
35.7 | - |
acetophenone | cofactor NADH, reduction, recombinant mutant G37Dm 30°C, pH 7.0 | Levilactobacillus brevis | |
36.6 | - |
NADH | oxidation, recombinant mutant G37D, 30°C, pH 7.0 | Levilactobacillus brevis | |
38.1 | - |
NADPH | oxidation, recombinant wild-type, 30°C, pH 7.0 | Levilactobacillus brevis | |
44.5 | - |
acetophenone | cofactor NADPH, reduction, recombinant wild-type, 30°C, pH 7.0 | Levilactobacillus brevis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
- |
Levilactobacillus brevis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Levilactobacillus brevis | |
NADH | - |
Levilactobacillus brevis | |
NADP+ | - |
Levilactobacillus brevis | |
NADPH | - |
Levilactobacillus brevis |