Short-chain alcohol dehydrogenase from Thermus thermophilus HB27
Enzyme Description
Species
Extremophile
Yes
"extremely thermophilic" organism
EC Number
Sequence
MGLFAGKGVLVTGGARGIGRAIAQAFAREGALVALCDLRPEGKEVAEAIGGAFFQVDLEDERERVRFVEEAAYALGRVDVLVNNAAIAAPGSALTVRLPEWRRVLEVNLTAPMHLSALAAREMRKVGGGAIVNVASVQGLFAEQENAAYNASKGGLVNLTRSLALDLAPLRIRVNAVAPGAIATEAVLEAIALSPDPERTRRDWEDLHALRRLGKPEEVAEAVLFLASEKASFITGAILPVDGGMTASFMMAGRPV
Angela Pennacchio et al. (2008)
β Purification and Characterization of a Novel Recombinant Highly Enantioselective Short-Chain NAD(H)-Dependent Alcohol Dehydrogenase from Thermus thermophilus
Applied and Environmental Microbiology
Β· doi:10.1128/AEM.00217-08 β
Β· Activity + Stability - Incubation
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Database ID
UniProt: A0A3P4ASV8 β
Sequence Annotation
Explicit - Provided
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)
Experimental Data (24 measurements)
24 measurements β page 1 of 2
| Property | Assay | Solvent | Solvent Volume | Aqueous Reference | Measured Value | Units | Extraction method | Data type | Solution | pH | Temperature | Substrate(s) | Product(s) | Cofactor(s) | Shaking | Comments |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Activity + Stability - Incubation | Activity after incubation (5 hours at 50Β°C), measured by absorbance spectrophotometry (change in NADH absorbance measurement, 340 nm) in the presence of organic solvent | Acetonitrile | 5 % v/v | 130 | 131 | % | Digital | Continuous | Incubation: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl, Assay: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl | Incubation: 6, Assay: 6 | Incubation: 50Β°C, Assay: 65Β°C | 20 mM Ethyl benzoylformate | Ethyl (R)-mandelate (95% ee) | 0.3 mM NADH | β | Water-incubated control (in %), activity measured in the presence of organic solvent | Not completely sure if the assay in the presence of organic solvent ("The concentration of the additive during the standard assay was as follows: 10 mM inhibitor, 10% detergent, and 50% (v/v) or 90% (v/v) organic solvent.") |
| Activity + Stability - Incubation | Activity after incubation (24 hours at 60Β°C), measured by absorbance spectrophotometry (change in NADH absorbance measurement, 340 nm) in the presence of organic solvent | Ethyl Acetate | 5 % v/v | 134 | 145 | % | Digital | Continuous | Incubation: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl, Assay: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl | Incubation: 6, Assay: 6 | Incubation: 60Β°C, Assay: 65Β°C | 20 mM Ethyl benzoylformate | Ethyl (R)-mandelate (95% ee) | 0.3 mM NADH | β | Water-incubated control (in %), activity measured in the presence of organic solvent | Not completely sure if the assay in the presence of organic solvent ("The concentration of the additive during the standard assay was as follows: 10 mM inhibitor, 10% detergent, and 50% (v/v) or 90% (v/v) organic solvent.") |
| Activity + Stability - Incubation | Activity after incubation (24 hours at 50Β°C), measured by absorbance spectrophotometry (change in NADH absorbance measurement, 340 nm) in the presence of organic solvent | Ethyl Acetate | 5 % v/v | 129 | 176 | % | Digital | Continuous | Incubation: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl, Assay: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl | Incubation: 6, Assay: 6 | Incubation: 60Β°C, Assay: 65Β°C | 20 mM Ethyl benzoylformate | Ethyl (R)-mandelate (95% ee) | 0.3 mM NADH | β | Water-incubated control (in %), activity measured in the presence of organic solvent | Not completely sure if the assay in the presence of organic solvent ("The concentration of the additive during the standard assay was as follows: 10 mM inhibitor, 10% detergent, and 50% (v/v) or 90% (v/v) organic solvent.") |
| Activity + Stability - Incubation | Activity after incubation (5 hours at 60Β°C), measured by absorbance spectrophotometry (change in NADH absorbance measurement, 340 nm) in the presence of organic solvent | Ethyl Acetate | 5 % v/v | 142 | 183 | % | Digital | Continuous | Incubation: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl, Assay: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl | Incubation: 6, Assay: 6 | Incubation: 50Β°C, Assay: 65Β°C | 20 mM Ethyl benzoylformate | Ethyl (R)-mandelate (95% ee) | 0.3 mM NADH | β | Water-incubated control (in %), activity measured in the presence of organic solvent | Not completely sure if the assay in the presence of organic solvent ("The concentration of the additive during the standard assay was as follows: 10 mM inhibitor, 10% detergent, and 50% (v/v) or 90% (v/v) organic solvent.") |
| Activity + Stability - Incubation | Activity after incubation (5 hours at 50Β°C), measured by absorbance spectrophotometry (change in NADH absorbance measurement, 340 nm) in the presence of organic solvent | Ethyl Acetate | 5 % v/v | 130 | 151 | % | Digital | Continuous | Incubation: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl, Assay: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl | Incubation: 6, Assay: 6 | Incubation: 50Β°C, Assay: 65Β°C | 20 mM Ethyl benzoylformate | Ethyl (R)-mandelate (95% ee) | 0.3 mM NADH | β | Water-incubated control (in %), activity measured in the presence of organic solvent | Not completely sure if the assay in the presence of organic solvent ("The concentration of the additive during the standard assay was as follows: 10 mM inhibitor, 10% detergent, and 50% (v/v) or 90% (v/v) organic solvent.") |
| Activity + Stability - Incubation | Activity after incubation (24 hours at 60Β°C), measured by absorbance spectrophotometry (change in NADH absorbance measurement, 340 nm) in the presence of organic solvent | 1,4-Dioxane | 5 % v/v | 134 | 161 | % | Digital | Continuous | Incubation: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl, Assay: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl | Incubation: 6, Assay: 6 | Incubation: 60Β°C, Assay: 65Β°C | 20 mM Ethyl benzoylformate | Ethyl (R)-mandelate (95% ee) | 0.3 mM NADH | β | Water-incubated control (in %), activity measured in the presence of organic solvent | Not completely sure if the assay in the presence of organic solvent ("The concentration of the additive during the standard assay was as follows: 10 mM inhibitor, 10% detergent, and 50% (v/v) or 90% (v/v) organic solvent.") |
| Activity + Stability - Incubation | Activity after incubation (24 hours at 50Β°C), measured by absorbance spectrophotometry (change in NADH absorbance measurement, 340 nm) in the presence of organic solvent | 1,4-Dioxane | 5 % v/v | 129 | 178 | % | Digital | Continuous | Incubation: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl, Assay: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl | Incubation: 6, Assay: 6 | Incubation: 60Β°C, Assay: 65Β°C | 20 mM Ethyl benzoylformate | Ethyl (R)-mandelate (95% ee) | 0.3 mM NADH | β | Water-incubated control (in %), activity measured in the presence of organic solvent | Not completely sure if the assay in the presence of organic solvent ("The concentration of the additive during the standard assay was as follows: 10 mM inhibitor, 10% detergent, and 50% (v/v) or 90% (v/v) organic solvent.") |
| Activity + Stability - Incubation | Activity after incubation (5 hours at 60Β°C), measured by absorbance spectrophotometry (change in NADH absorbance measurement, 340 nm) in the presence of organic solvent | 1,4-Dioxane | 5 % v/v | 142 | 180 | % | Digital | Continuous | Incubation: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl, Assay: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl | Incubation: 6, Assay: 6 | Incubation: 50Β°C, Assay: 65Β°C | 20 mM Ethyl benzoylformate | Ethyl (R)-mandelate (95% ee) | 0.3 mM NADH | β | Water-incubated control (in %), activity measured in the presence of organic solvent | Not completely sure if the assay in the presence of organic solvent ("The concentration of the additive during the standard assay was as follows: 10 mM inhibitor, 10% detergent, and 50% (v/v) or 90% (v/v) organic solvent.") |
| Activity + Stability - Incubation | Activity after incubation (5 hours at 50Β°C), measured by absorbance spectrophotometry (change in NADH absorbance measurement, 340 nm) in the presence of organic solvent | 1,4-Dioxane | 5 % v/v | 130 | 152 | % | Digital | Continuous | Incubation: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl, Assay: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl | Incubation: 6, Assay: 6 | Incubation: 50Β°C, Assay: 65Β°C | 20 mM Ethyl benzoylformate | Ethyl (R)-mandelate (95% ee) | 0.3 mM NADH | β | Water-incubated control (in %), activity measured in the presence of organic solvent | Not completely sure if the assay in the presence of organic solvent ("The concentration of the additive during the standard assay was as follows: 10 mM inhibitor, 10% detergent, and 50% (v/v) or 90% (v/v) organic solvent.") |
| Activity + Stability - Incubation | Activity after incubation (24 hours at 60Β°C), measured by absorbance spectrophotometry (change in NADH absorbance measurement, 340 nm) in the presence of organic solvent | Acetonitrile | 5 % v/v | 134 | 178 | % | Digital | Continuous | Incubation: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl, Assay: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl | Incubation: 6, Assay: 6 | Incubation: 60Β°C, Assay: 65Β°C | 20 mM Ethyl benzoylformate | Ethyl (R)-mandelate (95% ee) | 0.3 mM NADH | β | Water-incubated control (in %), activity measured in the presence of organic solvent | Not completely sure if the assay in the presence of organic solvent ("The concentration of the additive during the standard assay was as follows: 10 mM inhibitor, 10% detergent, and 50% (v/v) or 90% (v/v) organic solvent.") |
| Activity + Stability - Incubation | Activity after incubation (24 hours at 50Β°C), measured by absorbance spectrophotometry (change in NADH absorbance measurement, 340 nm) in the presence of organic solvent | Acetonitrile | 5 % v/v | 129 | 167 | % | Digital | Continuous | Incubation: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl, Assay: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl | Incubation: 6, Assay: 6 | Incubation: 60Β°C, Assay: 65Β°C | 20 mM Ethyl benzoylformate | Ethyl (R)-mandelate (95% ee) | 0.3 mM NADH | β | Water-incubated control (in %), activity measured in the presence of organic solvent | Not completely sure if the assay in the presence of organic solvent ("The concentration of the additive during the standard assay was as follows: 10 mM inhibitor, 10% detergent, and 50% (v/v) or 90% (v/v) organic solvent.") |
| Activity + Stability - Incubation | Activity after incubation (5 hours at 60Β°C), measured by absorbance spectrophotometry (change in NADH absorbance measurement, 340 nm) in the presence of organic solvent | Acetonitrile | 5 % v/v | 142 | 158 | % | Digital | Continuous | Incubation: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl, Assay: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl | Incubation: 6, Assay: 6 | Incubation: 50Β°C, Assay: 65Β°C | 20 mM Ethyl benzoylformate | Ethyl (R)-mandelate (95% ee) | 0.3 mM NADH | β | Water-incubated control (in %), activity measured in the presence of organic solvent | Not completely sure if the assay in the presence of organic solvent ("The concentration of the additive during the standard assay was as follows: 10 mM inhibitor, 10% detergent, and 50% (v/v) or 90% (v/v) organic solvent.") |
| Activity + Stability - Incubation | Activity after incubation (5 hours at 50Β°C), measured by absorbance spectrophotometry (change in NADH absorbance measurement, 340 nm) in the presence of organic solvent | Methanol | 5 % v/v | 130 | 138 | % | Digital | Continuous | Incubation: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl, Assay: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl | Incubation: 6, Assay: 6 | Incubation: 50Β°C, Assay: 65Β°C | 20 mM Ethyl benzoylformate | Ethyl (R)-mandelate (95% ee) | 0.3 mM NADH | β | Water-incubated control (in %), activity measured in the presence of organic solvent | Not completely sure if the assay in the presence of organic solvent ("The concentration of the additive during the standard assay was as follows: 10 mM inhibitor, 10% detergent, and 50% (v/v) or 90% (v/v) organic solvent.") |
| Activity + Stability - Incubation | Activity after incubation (24 hours at 60Β°C), measured by absorbance spectrophotometry (change in NADH absorbance measurement, 340 nm) in the presence of organic solvent | Isopropanol | 10 % v/v | 134 | 174 | % | Digital | Continuous | Incubation: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl, Assay: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl | Incubation: 6, Assay: 6 | Incubation: 60Β°C, Assay: 65Β°C | 20 mM Ethyl benzoylformate | Ethyl (R)-mandelate (95% ee) | 0.3 mM NADH | β | Water-incubated control (in %), activity measured in the presence of organic solvent | Not completely sure if the assay in the presence of organic solvent ("The concentration of the additive during the standard assay was as follows: 10 mM inhibitor, 10% detergent, and 50% (v/v) or 90% (v/v) organic solvent.") |
| Activity + Stability - Incubation | Activity after incubation (24 hours at 50Β°C), measured by absorbance spectrophotometry (change in NADH absorbance measurement, 340 nm) in the presence of organic solvent | Isopropanol | 10 % v/v | 129 | 211 | % | Digital | Continuous | Incubation: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl, Assay: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl | Incubation: 6, Assay: 6 | Incubation: 60Β°C, Assay: 65Β°C | 20 mM Ethyl benzoylformate | Ethyl (R)-mandelate (95% ee) | 0.3 mM NADH | β | Water-incubated control (in %), activity measured in the presence of organic solvent | Not completely sure if the assay in the presence of organic solvent ("The concentration of the additive during the standard assay was as follows: 10 mM inhibitor, 10% detergent, and 50% (v/v) or 90% (v/v) organic solvent.") |
| Activity + Stability - Incubation | Activity after incubation (5 hours at 60Β°C), measured by absorbance spectrophotometry (change in NADH absorbance measurement, 340 nm) in the presence of organic solvent | Isopropanol | 10 % v/v | 142 | 180 | % | Digital | Continuous | Incubation: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl, Assay: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl | Incubation: 6, Assay: 6 | Incubation: 50Β°C, Assay: 65Β°C | 20 mM Ethyl benzoylformate | Ethyl (R)-mandelate (95% ee) | 0.3 mM NADH | β | Water-incubated control (in %), activity measured in the presence of organic solvent | Not completely sure if the assay in the presence of organic solvent ("The concentration of the additive during the standard assay was as follows: 10 mM inhibitor, 10% detergent, and 50% (v/v) or 90% (v/v) organic solvent.") |
| Activity + Stability - Incubation | Activity after incubation (5 hours at 50Β°C), measured by absorbance spectrophotometry (change in NADH absorbance measurement, 340 nm) in the presence of organic solvent | Isopropanol | 10 % v/v | 130 | 174 | % | Digital | Continuous | Incubation: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl, Assay: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl | Incubation: 6, Assay: 6 | Incubation: 50Β°C, Assay: 65Β°C | 20 mM Ethyl benzoylformate | Ethyl (R)-mandelate (95% ee) | 0.3 mM NADH | β | Water-incubated control (in %), activity measured in the presence of organic solvent | Not completely sure if the assay in the presence of organic solvent ("The concentration of the additive during the standard assay was as follows: 10 mM inhibitor, 10% detergent, and 50% (v/v) or 90% (v/v) organic solvent.") |
| Activity + Stability - Incubation | Activity after incubation (24 hours at 60Β°C), measured by absorbance spectrophotometry (change in NADH absorbance measurement, 340 nm) in the presence of organic solvent | Isopropanol | 2 % v/v | 134 | 168 | % | Digital | Continuous | Incubation: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl, Assay: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl | Incubation: 6, Assay: 6 | Incubation: 60Β°C, Assay: 65Β°C | 20 mM Ethyl benzoylformate | Ethyl (R)-mandelate (95% ee) | 0.3 mM NADH | β | Water-incubated control (in %), activity measured in the presence of organic solvent | Not completely sure if the assay in the presence of organic solvent ("The concentration of the additive during the standard assay was as follows: 10 mM inhibitor, 10% detergent, and 50% (v/v) or 90% (v/v) organic solvent.") |
| Activity + Stability - Incubation | Activity after incubation (24 hours at 50Β°C), measured by absorbance spectrophotometry (change in NADH absorbance measurement, 340 nm) in the presence of organic solvent | Isopropanol | 2 % v/v | 129 | 193 | % | Digital | Continuous | Incubation: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl, Assay: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl | Incubation: 6, Assay: 6 | Incubation: 60Β°C, Assay: 65Β°C | 20 mM Ethyl benzoylformate | Ethyl (R)-mandelate (95% ee) | 0.3 mM NADH | β | Water-incubated control (in %), activity measured in the presence of organic solvent | Not completely sure if the assay in the presence of organic solvent ("The concentration of the additive during the standard assay was as follows: 10 mM inhibitor, 10% detergent, and 50% (v/v) or 90% (v/v) organic solvent.") |
| Activity + Stability - Incubation | Activity after incubation (5 hours at 60Β°C), measured by absorbance spectrophotometry (change in NADH absorbance measurement, 340 nm) in the presence of organic solvent | Isopropanol | 2 % v/v | 142 | 151 | % | Digital | Continuous | Incubation: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl, Assay: 100 mM MES buffer, 5mM 2-mercaptoethanol, 100 mM KCl | Incubation: 6, Assay: 6 | Incubation: 50Β°C, Assay: 65Β°C | 20 mM Ethyl benzoylformate | Ethyl (R)-mandelate (95% ee) | 0.3 mM NADH | β | Water-incubated control (in %), activity measured in the presence of organic solvent | Not completely sure if the assay in the presence of organic solvent ("The concentration of the additive during the standard assay was as follows: 10 mM inhibitor, 10% detergent, and 50% (v/v) or 90% (v/v) organic solvent.") |
Visualization : Activity + Stability β Incubation
One bar per measurement. Colour = solvent, shade = solvent volume.