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 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.
Levy
I
I
The Kennedy Leigh Center for Horticultural Research and The Otto Warburg Center for Agricultural Biotechnology, The Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, Rehovot, Israel.
Shoseyov
O
O
D000595:Amino Acid Sequence; D001435:Bacteriophages / Q000235:genetics; D001483:Base Sequence; D001665:Binding Sites; D002482:Cellulose / Q000737:chemistry* / Q000378:metabolism; D003001:Cloning, Molecular; D016384:Consensus Sequence; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D006735:Horseradish Peroxidase / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D017422:Sequence Analysis, DNA
D004800:Enzymes, Immobilized; D010455:Peptides; D002482:Cellulose; D006735:Horseradish Peroxidase