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On‐fiber display of a functional peptide at sites distant from the cell surface using a long bacterionanofiber of a trimeric autotransporter adhesin
Author(s) -
Nakatani Hajime,
Kanie Junichi,
Hori Katsutoshi
Publication year - 2019
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.26857
Subject(s) - bacterial adhesin , immunoelectron microscopy , fiber , escherichia coli , biophysics , cell , chemistry , biology , biochemistry , gene , genetics , antibody , organic chemistry
In the cell surface display system, the distance of a surface‐displayed molecule from the cell surface should influence its functionality due to the interference by other surface structures. For the purpose of developing this distance‐variable surface display system, we utilized a long fibrous adhesin, Acinetobacter trimeric autotransporter adhesin (AtaA) of the strain Tol 5. We constructed His‐tagged full‐length and shorter AtaA fibers designed by N‐terminal deletion and expressed them in the ΔataA mutant. Immunoelectron microscopy clearly showed that they formed fibers on the cell surface and the His‐tag was displayed on the fiber tip located at fixed distances from the cell surface. N‐terminal deletion of AtaA shortened the distance between the His‐tag and the cell surface, as designed. Time‐course analyses of the cell‐to‐Ni‐Sepharose beads binding revealed that cells producing the longer fibers bound more rapidly to the beads. The His‐tagged AtaA derivatives were also displayed on Escherichia coli cells, and a similar tendency was shown; the His‐tag on the longer fiber was more functional than that on the shorter one. Thus, we developed an on‐fiber display system of a functional peptide using a long trimeric autotransporter adhesin (TAA) fiber, which can vary the distance between the displayed molecule and the cell surface.