Haloquadratum walsbyi yields a versatile, NAD+/NADP+ dual affinity, thermostable, alcohol dehydrogenase (HwADH)

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 op...

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Main Authors: Cassidy, Jennifer, Paradisi, Francesca
Format: Article
Published: Springer 2018
Subjects:
Online Access:https://eprints.nottingham.ac.uk/51157/
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author Cassidy, Jennifer
Paradisi, Francesca
author_facet Cassidy, Jennifer
Paradisi, Francesca
author_sort Cassidy, Jennifer
building Nottingham Research Data Repository
collection Online Access
description 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, 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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spelling nottingham-511572020-05-04T19:50:51Z https://eprints.nottingham.ac.uk/51157/ Haloquadratum walsbyi yields a versatile, NAD+/NADP+ dual affinity, thermostable, alcohol dehydrogenase (HwADH) Cassidy, Jennifer Paradisi, Francesca 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, 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. Springer 2018-06 Article PeerReviewed Cassidy, Jennifer and Paradisi, Francesca (2018) Haloquadratum walsbyi yields a versatile, NAD+/NADP+ dual affinity, thermostable, alcohol dehydrogenase (HwADH). Molecular Biotechnology, 60 (6). pp. 420-426. ISSN 1559-0305 Haloquadratum walsbyi; Alcohol dehydrogenase; Thermoactivity; Dual cofactor specificity https://link.springer.com/article/10.1007%2Fs12033-018-0083-6 doi:10.1007/s12033-018-0083-6 doi:10.1007/s12033-018-0083-6
spellingShingle Haloquadratum walsbyi; Alcohol dehydrogenase; Thermoactivity; Dual cofactor specificity
Cassidy, Jennifer
Paradisi, Francesca
Haloquadratum walsbyi yields a versatile, NAD+/NADP+ dual affinity, thermostable, alcohol dehydrogenase (HwADH)
title Haloquadratum walsbyi yields a versatile, NAD+/NADP+ dual affinity, thermostable, alcohol dehydrogenase (HwADH)
title_full Haloquadratum walsbyi yields a versatile, NAD+/NADP+ dual affinity, thermostable, alcohol dehydrogenase (HwADH)
title_fullStr Haloquadratum walsbyi yields a versatile, NAD+/NADP+ dual affinity, thermostable, alcohol dehydrogenase (HwADH)
title_full_unstemmed Haloquadratum walsbyi yields a versatile, NAD+/NADP+ dual affinity, thermostable, alcohol dehydrogenase (HwADH)
title_short Haloquadratum walsbyi yields a versatile, NAD+/NADP+ dual affinity, thermostable, alcohol dehydrogenase (HwADH)
title_sort haloquadratum walsbyi yields a versatile, nad+/nadp+ dual affinity, thermostable, alcohol dehydrogenase (hwadh)
topic Haloquadratum walsbyi; Alcohol dehydrogenase; Thermoactivity; Dual cofactor specificity
url https://eprints.nottingham.ac.uk/51157/
https://eprints.nottingham.ac.uk/51157/
https://eprints.nottingham.ac.uk/51157/