Characterization of alcohol dehydrogenase (ADH12) from Haloarcula marismortui, an extreme halophile from the Dead Sea
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|Title:||Characterization of alcohol dehydrogenase (ADH12) from Haloarcula marismortui, an extreme halophile from the Dead Sea||Authors:||Timpson, Leanne M.
Mac Donnchadha, Cillin
Sharkey, Michael A.
|Permanent link:||http://hdl.handle.net/10197/3635||Date:||Jan-2012||Online since:||2012-06-12T16:34:10Z||Abstract:||Haloarchaeal alcohol dehydrogenases are of increasing interest as biocatalysts in the field of white biotechnology. In this study, the gene adh12 from the extreme halophile Haloarcula marismortui (HmADH12), encoding a 384 residue protein, was cloned into two vectors: pRV1 and pTA963. The resulting constructs were used to transform host strains Haloferax volcanii (DS70) and (H1209), respectively. Overexpressed His-tagged recombinant HmADH12 was purified by immobilized metal-affinity chromatography (IMAC). The His-tagged protein was visualized by SDS-PAGE, with a subunit molecular mass of 41.6 kDa, and its identity was confirmed by mass spectrometry. Purified HmADH12 catalyzed the interconversion between alcohols and aldehydes and ketones, being optimally active in the presence of 2 M KCl. It was thermoactive, with maximum activity registered at 60°C. The NADP(H) dependent enzyme was haloalkaliphilic for the oxidative reaction with optimum activity at pH 10.0. It favored a slightly acidic pH of 6.0 for catalysis of the reductive reaction. HmADH12 was significantly more tolerant than mesophilic ADHs to selected organic solvents, making it a much more suitable biocatalyst for industrial application.||Funding Details:||Science Foundation Ireland
Irish Research Council for Science, Engineering and Technology
|Type of material:||Journal Article||Publisher:||Springer||Journal:||Extremophiles||Volume:||16||Issue:||1||Start page:||57||End page:||66||Copyright (published version):||2012 Springer||Keywords:||Alcohol dehydrogenase; Biocatalysis; Extremophile; Halophile; Haloarcula marismortui; Haloferax volcanii; Organic solvents||Subject LCSH:||Alcohol dehydrogenase
|DOI:||10.1007/s00792-011-0405-0||Language:||en||Status of Item:||Peer reviewed|
|Appears in Collections:||Biomolecular and Biomedical Science Research Collection|
Chemistry Research Collection
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