Covalent immobilization of alcohol dehydrogenase (ADH2) from Haloferax volcanii: how to maximize activity and optimize performance of halophilic enzymes

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Title: Covalent immobilization of alcohol dehydrogenase (ADH2) from Haloferax volcanii: how to maximize activity and optimize performance of halophilic enzymes
Authors: Alsafadi, Diya
Paradisi, Francesca
Permanent link: http://hdl.handle.net/10197/5423
Date: Mar-2014
Abstract: 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 Sepaeads 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.
Funding Details: Science Foundation Ireland
Type of material: Journal Article
Publisher: Springer
Journal: Molecular Biotechnology
Volume: 56
Issue: 3
Start page: 240
End page: 247
Copyright (published version): 2014, Springer
Keywords: Enzyme immobilizationSepabeadsHalophilic enzymeAlcohol dehydrogenaseHaloferax volcaniiOrganic solvents tolerance
DOI: 10.1007/s12033-013-9701-5
Language: en
Status of Item: Peer reviewed
Appears in Collections:Chemistry Research Collection

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