Crystallization (Comment) | Organism |
---|---|
crystal structure of the apoenzyme as well as its NADP+-bound state with resolutions down to 2.8 A. ADH displays a homotetrameric quaternary structure that can be described as a dimer of homodimers while in each subunit a seven-stranded parallel beta-sheet, flanked by three alpha-helices on each side, forms a Rossmann fold-type dinucleotide binding domain. NADP+ specificity is largely governed by the residues Asn15, Gly37, Arg38, and Arg39 | Ralstonia sp. |
Protein Variants | Comment | Organism |
---|---|---|
G37D | mutation in NADP+-binding site, strong decrease in activity with NADP+ | Ralstonia sp. |
N15G/G37D/R38V/R39S | mutation in NADP+-binding site, strong decrease in activity with NADP+ | Ralstonia sp. |
N15G/G37D/R38V/R39S/A86N/S88A | mutation in NADP+-binding site, strong decrease in activity with NADP+ | Ralstonia sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.051 | - |
NADP+ | wild-type, pH 8.3, 30°C | Ralstonia sp. | |
0.28 | - |
NADP+ | mutant N15G/G37D/R38V/R39S, pH 8.3, 30°C | Ralstonia sp. | |
0.8 | - |
NAD+ | mutant N15G/G37D/R38V/R39S/A86N/S88A, pH 8.3, 30°C | Ralstonia sp. | |
1.1 | - |
NAD+ | mutant N15G/G37D/R38V/R39S, pH 8.3, 30°C | Ralstonia sp. | |
1.2 | - |
(S)-1-phenylethanol | mutant G37D, pH 8.3, 30°C | Ralstonia sp. | |
2.3 | - |
NAD+ | mutant G37D, pH 8.3, 30°C | Ralstonia sp. | |
2.3 | - |
NADP+ | mutant G37D, pH 8.3, 30°C | Ralstonia sp. | |
3.1 | - |
NAD+ | wild-type, pH 8.3, 30°C | Ralstonia sp. | |
5.5 | - |
(R)-1-phenylethanol | wild-type, pH 8.3, 30°C | Ralstonia sp. | |
8.3 | - |
(S)-1-phenylethanol | mutant N15G/G37D/R38V/R39S, pH 8.3, 30°C | Ralstonia sp. | |
9.9 | - |
(S)-1-phenylethanol | wild-type, pH 8.3, 30°C | Ralstonia sp. | |
17.6 | - |
(S)-1-phenylethanol | mutant N15G/G37D/R38V/R39S/A86N/S88A, pH 8.3, 30°C | Ralstonia sp. |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Ralstonia sp. | C0IR58 | - |
- |
Ralstonia sp. DSM Z 6428 | C0IR58 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(R)-1-phenylethanol + NAD+ | - |
Ralstonia sp. | 1-phenylacetaldehyde + NADH + H+ | - |
? | |
(R)-1-phenylethanol + NAD+ | - |
Ralstonia sp. DSM Z 6428 | 1-phenylacetaldehyde + NADH + H+ | - |
? | |
(S)-1-phenylethanol + NADP+ | - |
Ralstonia sp. | 1-phenylacetaldehyde + NADPH + H+ | - |
? | |
(S)-1-phenylethanol + NADP+ | - |
Ralstonia sp. DSM Z 6428 | 1-phenylacetaldehyde + NADPH + H+ | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00003 | - |
NADP+ | mutant N15G/G37D/R38V/R39S, pH 8.3, 30°C | Ralstonia sp. | |
0.0025 | - |
NADP+ | mutant G37D, pH 8.3, 30°C | Ralstonia sp. | |
0.0047 | - |
(R)-1-phenylethanol | wild-type, pH 8.3, 30°C | Ralstonia sp. | |
0.0115 | - |
(S)-1-phenylethanol | wild-type, pH 8.3, 30°C | Ralstonia sp. | |
0.022 | - |
NAD+ | wild-type, pH 8.3, 30°C | Ralstonia sp. | |
0.023 | - |
(S)-1-phenylethanol | mutant N15G/G37D/R38V/R39S/A86N/S88A, pH 8.3, 30°C | Ralstonia sp. | |
0.03 | - |
NAD+ | mutant N15G/G37D/R38V/R39S/A86N/S88A, pH 8.3, 30°C | Ralstonia sp. | |
0.038 | - |
NAD+ | mutant G37D, pH 8.3, 30°C | Ralstonia sp. | |
0.042 | - |
(S)-1-phenylethanol | mutant N15G/G37D/R38V/R39S, pH 8.3, 30°C | Ralstonia sp. | |
0.08 | - |
NAD+ | mutant N15G/G37D/R38V/R39S, pH 8.3, 30°C | Ralstonia sp. | |
0.783 | - |
(S)-1-phenylethanol | mutant G37D, pH 8.3, 30°C | Ralstonia sp. | |
1.65 | - |
NADP+ | wild-type, pH 8.3, 30°C | Ralstonia sp. |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0001 | - |
NADP+ | mutant N15G/G37D/R38V/R39S, pH 8.3, 30°C | Ralstonia sp. | |
0.0011 | - |
NADP+ | mutant G37D, pH 8.3, 30°C | Ralstonia sp. | |
0.00115 | - |
(S)-1-phenylethanol | wild-type, pH 8.3, 30°C | Ralstonia sp. | |
0.0013 | - |
(S)-1-phenylethanol | mutant N15G/G37D/R38V/R39S/A86N/S88A, pH 8.3, 30°C | Ralstonia sp. | |
0.0047 | - |
(R)-1-phenylethanol | wild-type, pH 8.3, 30°C | Ralstonia sp. | |
0.0052 | - |
(S)-1-phenylethanol | mutant N15G/G37D/R38V/R39S, pH 8.3, 30°C | Ralstonia sp. | |
0.007 | - |
NAD+ | wild-type, pH 8.3, 30°C | Ralstonia sp. | |
0.0167 | - |
NAD+ | mutant G37D, pH 8.3, 30°C | Ralstonia sp. | |
0.038 | - |
NAD+ | mutant N15G/G37D/R38V/R39S/A86N/S88A, pH 8.3, 30°C | Ralstonia sp. | |
0.073 | - |
NAD+ | mutant N15G/G37D/R38V/R39S, pH 8.3, 30°C | Ralstonia sp. | |
0.65 | - |
(S)-1-phenylethanol | mutant G37D, pH 8.3, 30°C | Ralstonia sp. | |
32.33 | - |
NADP+ | wild-type, pH 8.3, 30°C | Ralstonia sp. |