Abstract
This study presents the first example of an alcohol dehydrogenase (ADH) from the halophilic archaeum Haloquadratum walsbyi (HwADH). A hexahistidine-tagged recombinant HwADH was heterologously overexpressed in Haloferax volcanii. HwADH was purified in one step and was found to be thermophilic with optimal activity at 65 °C. HwADH was active in the presence of 10% (v/v) organic solvent. The enzyme displayed dual cofactor specificity and a broad substrate scope, and maximum activity was detected with benzyl alcohol and 2-phenyl-1-propanol. HwADH accepted aromatic ketones, acetophenone and phenylacetone as substrates. The enzyme also accepted cyclohexanol and aromatic secondary alcohols, 1-phenylethanol and 4-phenyl-2-butanol. H. walsbyi may offer an excellent alternative to other archaeal sources to expand the toolbox of halophilic biocatalysts.
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Abbreviations
- ACN:
-
Acetonitrile
- ADH:
-
Alcohol dehydrogenase
- IMAC:
-
Immobilised metal-affinity chromatography
- iPrOH:
-
Isopropanol
- SDS–PAGE:
-
Sodium dodecyl sulphate–polyacrylamide gel electrophoresis
- EtOH:
-
Ethanol
- BzOH:
-
Benzyl alcohol
- CycOH:
-
Cyclohexanol
- 1-PheOH:
-
1-Phenylethanol
- (S)-1-PheOH:
-
(S)-1-phenylethanol
- (R)-1-PheOH:
-
(R)-1-phenylethanol
- 2-Phe-1-Prop:
-
2-Phenyl-1-propanol
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Acknowledgements
The authors wish to acknowledge the support from the Synthesis and Solid State Pharmaceutical Centre and Science Foundation Ireland, Grant Number 12/RC/2275, and Dr. Thorsten Allers for useful discussions.
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Cassidy, J., Paradisi, F. Haloquadratum walsbyi Yields a Versatile, NAD+/NADP+ Dual Affinity, Thermostable, Alcohol Dehydrogenase (HwADH). Mol Biotechnol 60, 420–426 (2018). https://doi.org/10.1007/s12033-018-0083-6
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DOI: https://doi.org/10.1007/s12033-018-0083-6