| Title |
Covalent Immobilization of Alcohol Dehydrogenase (ADH2) from Haloferax volcanii: How to Maximize Activity and Optimize Performance of Halophilic Enzymes
|
|---|---|
| Published in |
Molecular Biotechnology, September 2013
|
| DOI | 10.1007/s12033-013-9701-5 |
| Pubmed ID | |
| Authors |
Diya Alsafadi, Francesca Paradisi |
| 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 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. |
Mendeley readers
The data shown below were compiled from readership statistics for 55 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Mexico | 1 | 2% |
| Unknown | 54 | 98% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 12 | 22% |
| Student > Ph. D. Student | 12 | 22% |
| Student > Doctoral Student | 5 | 9% |
| Student > Bachelor | 4 | 7% |
| Student > Master | 3 | 5% |
| Other | 7 | 13% |
| Unknown | 12 | 22% |
| Readers by discipline | Count | As % |
|---|---|---|
| Chemistry | 13 | 24% |
| Chemical Engineering | 8 | 15% |
| Agricultural and Biological Sciences | 8 | 15% |
| Biochemistry, Genetics and Molecular Biology | 7 | 13% |
| Engineering | 2 | 4% |
| Other | 4 | 7% |
| Unknown | 13 | 24% |