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Expression, refolding and indirect immobilization of horseradish peroxidase (HRP) to cellulose via a phage‐selected peptide and cellulose‐binding domain (CBD)
Author(s) -
Levy Ilan,
Shoseyov Oded
Publication year - 2001
Publication title -
journal of peptide science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 66
eISSN - 1099-1387
pISSN - 1075-2617
DOI - 10.1002/psc.294
Subject(s) - horseradish peroxidase , peptide , cellulose , chemistry , peptide library , phage display , combinatorial chemistry , microcrystalline cellulose , biochemistry , peptide sequence , enzyme , gene
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.