Expression, refolding and indirect immobilization of horseradish peroxidase (HRP) to cellulose via a phage-selected peptide and cellulose-binding domain (CBD)
✍ Scribed by Ilan Levy; Oded Shoseyov
- Book ID
- 105360062
- Publisher
- John Wiley and Sons
- Year
- 2001
- Tongue
- English
- Weight
- 117 KB
- Volume
- 7
- Category
- Article
- ISSN
- 1075-2617
- DOI
- 10.1002/psc.294
No coin nor oath required. For personal study only.
✦ Synopsis
Abstract
We examined the potential immobilization of horseradish peroxidase (HRP) to cellulose with cellulose‐binding domain (CBD) as a mediator, using a ligand selected from a phage‐displayed random peptide library. A 15‐mer random peptide library was panned on cellulose‐coated plates covered with CBD in order to find a peptide that binds to CBD in its bound form. The sequence I/LHS, which was found to be an efficient binder of CBD, was fused to a synthetic gene of HRP as an affinity tag. The tagged enzyme (tHRP) was then immobilized on microcrystalline cellulose coated with CBD, thereby demonstrating the indirect immobilization of a protein to cellulose via three amino acids selected by phage display library and CBD. Copyright © 2001 European Peptide Society and John Wiley & Sons, Ltd.