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Literature summary for 1.1.1.1 extracted from
- Picosecond-resolved fluorescence studies of substrate and cofactor-binding domain mutants in a thermophilic alcohol dehydrogenase uncover an extended network of communication (2014), J. Am. Chem. Soc., 136, 14821-14833.
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| additional information | mutations Y25A (at the dimer interface) and V260A (at the cofactor-binding domain) exhibit opposing low-temperature effects on the hydride tunneling step. The distal Y25A increases active-site flexibility, V260A introduces a temperature-dependent equilibration process, and the double mutant (Y25A/V260A) eliminates the temperature-dependent transition sensed by the active-site tryptophan in the presence of V260A. V260A displays a structural change in the active-site environment/solvation | Geobacillus stearothermophilus |
| V260A | kinetic parameters and temperature dependencies similar to wild-type | Geobacillus stearothermophilus |
| W49F/W167Y | kinetic parameters and temperature dependencies similar to wild-type | Geobacillus stearothermophilus |
| W49F/W167Y/V260A | kinetic parameters and temperature dependencies similar to wild-type | Geobacillus stearothermophilus |
| W49F/W87F | kinetic parameters and temperature dependencies similar to wild-type | Geobacillus stearothermophilus |
| W49F/W87F/V260A | kinetic parameters and temperature dependencies similar to wild-type | Geobacillus stearothermophilus |
| Y25A | kinetic parameters and temperature dependencies similar to wild-type | Geobacillus stearothermophilus |
| Y25A/W49F/W167Y | kinetic parameters and temperature dependencies similar to wild-type | Geobacillus stearothermophilus |
| Y25A/W49F/W167Y/V260A | kinetic parameters and temperature dependencies similar to wild-type | Geobacillus stearothermophilus |
| Y25A/W49F/W87F | kinetic parameters and temperature dependencies similar to wild-type | Geobacillus stearothermophilus |
| Y25A/W49F/W87F/V260A | kinetic parameters and temperature dependencies similar to wild-type | Geobacillus stearothermophilus |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.5 | - |
NAD+ | mutant Y25A/W49F/W167Y, 30°C, pH not specified in the publication | Geobacillus stearothermophilus |  |
| 0.8 | - |
benzyl alcohol | mutant Y25A/W49F/W167Y, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 0.9 | - |
NAD+ | mutant Y25A/W49F/W87F, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 1 | - |
NAD+ | mutant Y25A, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 1.1 | - |
NAD+ | wild-type, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 1.3 | - |
NAD+ | mutant W49F/W87F, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 1.8 | - |
NAD+ | mutant W49F/W167Y, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 3.1 | - |
benzyl alcohol | mutant W49F/W167Y, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 4.2 | - |
benzyl alcohol | mutant V260A, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 5.4 | - |
benzyl alcohol | mutant Y25A/W49F/W167Y/V260A, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 6 | - |
benzyl alcohol | mutant W49F/W87F, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 6.8 | - |
benzyl alcohol | wild-type, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 8.2 | - |
benzyl alcohol | mutant W49F/W167Y/V260A, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 8.3 | - |
NAD+ | mutant W49F/W167Y/V260A, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 9.7 | - |
NAD+ | mutant Y25A/W49F/W167Y/V260A, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 10 | - |
NAD+ | mutant V260A, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 10.1 | - |
benzyl alcohol | mutant Y25A/W49F/W87F/V260A, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 10.2 | - |
NAD+ | mutant W49F/W87F/V260A, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 10.9 | - |
benzyl alcohol | mutant Y25A/W49F/W87F, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 12.9 | - |
benzyl alcohol | mutant W49F/W87F/V260A, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 14.8 | - |
NAD+ | mutant Y25A/W49F/W87F/V260A, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 16.5 | - |
benzyl alcohol | mutant Y25A, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Geobacillus stearothermophilus | P42328 | - |
- |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| benzyl alcohol + NAD+ | - |
Geobacillus stearothermophilus | benzaldehyde + NADH + H+ | - |
? | |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 1 | - |
NAD+ | mutant W49F/W87F/V260A, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 1.4 | - |
NAD+ | mutant W49F/W167Y/V260A, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 1.9 | - |
NAD+ | mutant V260A, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 1.9 | - |
NAD+ | mutant Y25A/W49F/W167Y/V260A, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 2.5 | - |
NAD+ | mutant Y25A/W49F/W167Y, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 2.5 | - |
NAD+ | mutant Y25A/W49F/W87F/V260A, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 3 | - |
NAD+ | mutant W49F/W87F, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 3.4 | - |
NAD+ | mutant W49F/W167Y, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 5.1 | - |
NAD+ | mutant Y25A, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 6.3 | - |
NAD+ | mutant Y25A/W49F/W87F, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
| 8 | - |
NAD+ | wild-type, 30°C, pH not specified in the publication | Geobacillus stearothermophilus | |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| NAD+ | - |
Geobacillus stearothermophilus | |
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