Molecular Biotechnology

, Volume 56, Issue 3, pp 240–247 | Cite as

Covalent Immobilization of Alcohol Dehydrogenase (ADH2) from Haloferax volcanii: How to Maximize Activity and Optimize Performance of Halophilic Enzymes

  • Diya Alsafadi
  • Francesca ParadisiEmail author


Alcohol dehydrogenase from halophilic archaeon Haloferax volcanii (HvADH2) was successfully covalently immobilized on metal-derivatized epoxy Sepabeads. The immobilization conditions were optimized by investigating several parameters that affect the halophilic enzyme–support interaction. The highest immobilization efficiency (100 %) and retention activity (60 %) were achieved after 48 h of incubation of the enzyme with Ni-epoxy Sepabeads support in 100 mM Tris–HCl buffer, pH 8, containing 3 M KCl at 5 °C. No significant stabilization was observed after blocking the unreacted epoxy groups with commonly used hydrophilic agents. A significant increase in the stability of the immobilized enzyme was achieved by blocking the unreacted epoxy groups with ethylamine. The immobilization process increased the enzyme stability, thermal activity, and organic solvents tolerance when compared to its soluble counterpart, indicating that the immobilization enhances the structural and conformational stability. One step purification–immobilization of this enzyme has been carried out on metal chelate-epoxy Sepabeads, as an efficient method to obtain immobilized biocatalyst directly from bacterial extracts.


Enzyme immobilization Sepabeads Halophilic enzyme Alcohol dehydrogenase Haloferax volcanii Organic solvents tolerance 



This work was supported by funding provided by the Islamic Development Bank (IDB) and by Science Foundation Ireland (SFI). The company Resindion S. R. L. (Milano, Italy) kindly donated the epoxy sepabeads.


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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  1. 1.Centre for Synthesis and Chemical Biology, UCD School of Chemistry and Chemical BiologyUniversity College DublinDublin 4Ireland

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