Levy I, Shoseyov O
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.
J Pept Sci. 2001 Jan;7(1):50-7. doi: 10.1002/psc.294.
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.
我们使用从噬菌体展示随机肽库中筛选出的一种配体，研究了以纤维素结合结构域（CBD）作为媒介，将辣根过氧化物酶（HRP）固定到纤维素上的可能性。为了找到能以结合形式与CBD结合的肽段，我们在覆盖有CBD的纤维素包被平板上筛选了一个15聚体随机肽库。发现序列I/LHS是一种有效的CBD结合剂，将其作为亲和标签融合到HRP的合成基因上。然后将带标签的酶（tHRP）固定在涂有CBD的微晶纤维素上，从而证明了通过噬菌体展示文库筛选出的三个氨基酸以及CBD，可将蛋白质间接固定到纤维素上。
