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F ine‐tuning sortase‐mediated immobilization of protein layers on surfaces using sequential deprotection and coupling
Author(s) -
RaeeszadehSarmazdeh Maryam,
Parthasarathy Ranganath,
Boder Eric T.
Publication year - 2017
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1002/btpr.2449
Subject(s) - sortase , sortase a , coupling (piping) , chemistry , biophysics , combinatorial chemistry , chemical engineering , materials science , biochemistry , bacterial protein , biology , composite material , gene , engineering
Increasing interest in protein immobilization on surfaces has heightened the need for techniques enabling layer‐by‐layer protein attachment. Here, we report a technique for controlling enzyme‐mediated immobilization of layers of protein on the surface using a genetically encoded protecting group. An enterokinase‐cleavable peptide sequence was inserted at the N‐terminus of bifunctional fluorescent proteins containing Sortase A substrate recognition tags at both ends to control Sortase A‐mediated protein immobilization on the surface layer‐by‐layer. Efficient, sequential immobilization of a second layer of protein using Sortase A required removal of the N‐terminal protecting group, suggesting the method enables multilayer synthesis using cyclic deprotection and coupling steps. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:824–831, 2017