Expression, refolding and indirect immobilization of horseradish peroxidase (HRP) to cellulose via a phage-selected peptide and cellulose-binding domain (CBD)

We examined the potential immobilization of horseradish peroxidase (HRP] to cellulose with cellulose-binding domain (CBD) as a mediator, using a ligan d selected from a phage-displayed random peptide library. A 15-mer random p eptide library was panned on cellulose-coated plates covered with CBD in or der to fmd a peptide that binds to CBD in its bound form. The sequence I/LH S, which was found to be an efficient binder of CBD, was fused to a synthet ic gene of HRP as an affinity tag. The tagged enzyme (tHRP) was then immobi lized 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 (C) 2001 European Pep tide Society and John Wiley & Sons, Ltd.