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  5. Heterologous overexpression, purification and characterisation of an alcohol dehydrogenase (ADH2) from Halobacterium sp. NRC-1
 
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Heterologous overexpression, purification and characterisation of an alcohol dehydrogenase (ADH2) from Halobacterium sp. NRC-1

Author(s)
Liliensiek, Ann-Kathrin  
Cassidy, Jennifer  
Gucciardo, Gabriele  
Whitely, Cliadhna  
Paradisi, Francesca  
Uri
http://hdl.handle.net/10197/5422
Date Issued
2013-10
Date Available
2014-10-01T03:00:19Z
Abstract
Replacement of chemical steps with biocatalytic ones is becoming increasingly more interesting due to the remarkable catalytic properties of enzymes, such as their wide range of substrate specificities and variety of chemo-, stereo- and regioselective reactions.
This study presents characterization of an alcohol dehydrogenase (ADH) from the halophilic archaeum Halobacterium sp. NRC-1 (HsADH2). A hexahistidine-tagged recombinant version of HsADH2 (His-HsADH2) was heterologously overexpressed in Haloferax volcanii. The enzyme was purified in one step by immobilised Ni-affinity chromatography (IMAC). His-HsADH2 was halophilic and mildly thermophilic with optimal activity for ethanol oxidation at 4 M KCl around 60 °C and pH 10.0. The enzyme was extremely stable, retaining 80 % activity after 30 days. His-HsADH2 showed preference for NADP(H) but interestingly retained 60 % activity towards NADH. The enzyme displayed broad substrate specificity, with maximum activity obtained for 1-propanol. The enzyme also accepted secondary alcohols such as 2-butanol and even 1-phenylethanol. In the reductive reaction, working conditions for His-HsADH2 were optimised for acetaldehyde and found to be 4 M KCl and pH 6.0. His-HsADH2 displayed intrinsic organic solvent tolerance, which is highly relevant for biotechnological applications.
Sponsorship
Science Foundation Ireland
Other Sponsorship
EPA
Type of Material
Journal Article
Publisher
Springer
Journal
Molecular Biotechnology
Volume
55
Issue
2
Start Page
143
End Page
149
Copyright (Published Version)
2013, Springer
Subjects

Halobacterium sp. NRC...

Haloferax volcanii

Alcohol dehydrogenase...

Solvent tolerance

DOI
10.1007/s12033-013-9666-4
Language
English
Status of Item
Peer reviewed
This item is made available under a Creative Commons License
https://creativecommons.org/licenses/by-nc-nd/3.0/ie/
File(s)
No Thumbnail Available
Name

Liliensiek_Mol_Biotech_2013.doc

Size

265 KB

Format

Microsoft Word

Checksum (MD5)

002ac7a7e399489d709442b14a6e2614

Owning collection
Chemistry Research Collection

Item descriptive metadata is released under a CC-0 (public domain) license: https://creativecommons.org/public-domain/cc0/.
All other content is subject to copyright.

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