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Applied Microbiology and Biotechnology

, Volume 97, Issue 1, pp 195–203 | Cite as

A comparison of two novel alcohol dehydrogenase enzymes (ADH1 and ADH2) from the extreme halophile Haloferax volcanii

  • Leanne M. Timpson
  • Ann-Kathrin Liliensiek
  • Diya Alsafadi
  • Jennifer Cassidy
  • Michael A. Sharkey
  • Susan Liddell
  • Thorsten Allers
  • Francesca Paradisi
Biotechnologically relevant enzymes and proteins

Abstract

Haloarchaeal alcohol dehydrogenases are exciting biocatalysts with potential industrial applications. In this study, two alcohol dehydrogenase enzymes from the extremely halophilic archaeon Haloferax volcanii (HvADH1 and HvADH2) were homologously expressed and subsequently purified by immobilized metal-affinity chromatography. The proteins appeared to copurify with endogenous alcohol dehydrogenases, and a double Δadh2 Δadh1 gene deletion strain was constructed to prevent this occurrence. Purified HvADH1 and HvADH2 were compared in terms of stability and enzymatic activity over a range of pH values, salt concentrations, and temperatures. Both enzymes were haloalkaliphilic and thermoactive for the oxidative reaction and catalyzed the reductive reaction at a slightly acidic pH. While the NAD+-dependent HvADH1 showed a preference for short-chain alcohols and was inherently unstable, HvADH2 exhibited dual cofactor specificity, accepted a broad range of substrates, and, with respect to HvADH1, was remarkably stable. Furthermore, HvADH2 exhibited tolerance to organic solvents. HvADH2 therefore displays much greater potential as an industrially useful biocatalyst than HvADH1.

Keywords

Alcohol dehydrogenase Biocatalyst discovery Protein characterization Extremophile Haloferax volcanii Organic solvents 

Notes

Acknowledgments

This work was supported by a research grant awarded to F.P. by Science Foundation Ireland (SFI), a University Research Fellowship awarded to T.A. by the Royal Society, and funding provided by the Islamic Development Bank (IsDB).

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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Leanne M. Timpson
    • 1
  • Ann-Kathrin Liliensiek
    • 1
  • Diya Alsafadi
    • 1
  • Jennifer Cassidy
    • 1
  • Michael A. Sharkey
    • 2
  • Susan Liddell
    • 3
  • Thorsten Allers
    • 4
  • Francesca Paradisi
    • 1
  1. 1.Centre for Synthesis and Chemical Biology, UCD School of Chemistry and Chemical BiologyUniversity College DublinBelfieldIreland
  2. 2.UCD School of Biomolecular and Biomedical Science, Conway InstituteUniversity College DublinBelfieldIreland
  3. 3.Division of Animal Sciences, School of BiosciencesUniversity of NottinghamLoughboroughUK
  4. 4.Institute of Genetics, School of BiologyUniversity of Nottingham, Queen’s Medical CentreNottinghamUK

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