title: Enzyme directed formation of un-natural side-chains for covalent surface attachment of proteins.
abstract: The covalent immobilization of proteins onto surfaces is an essential aspect of several fields of research, including proteomics, sensing, heterogeneous biocatalysis, and more broadly biotechnology. Site-specific, covalent attachment of proteins has been achieved in recent years by the use of expanded genetic codes to produce proteins with controlled placement of un-natural amino acids bearing bio-orthogonal functional groups. Unfortunately, the complexity of developing such systems is impractical for most laboratories; hence, a less complicated approach to generating un-natural amino acid side-chains has been employed. Utilizing a straightforward reaction with formylglycine generating enzyme, we use the site-specific modification of engineered proteins to yield un-natural amino acid side-chains for protein immobilization. Using this approach, we demonstrate the controlled immobilization of various enzymes onto a variety of amine coated surfaces. Our results reveal reusability of the immobilized enzymes via this strategy, and furthermore, we find the activity of the immobilized enzymes to remain even after a month of use indicating significant stability of the linkage.
authors.affiliation: Department of Chemical Engineering, Hanyang University, 222 Wangsimni-ro, Seongdong-gu, Seoul 133-791, Republic of Korea; Institute of Nanoscience and Technology, Hanyang University, 222 Wangsimni-ro, Seongdong-gu, Seoul 133-791, Republic of Korea.
authors.forename: Hwayoung
authors.initials: H
authors.lastname: Cho
authors.affiliation: Department of Chemical Engineering, Hanyang University, 222 Wangsimni-ro, Seongdong-gu, Seoul 133-791, Republic of Korea; Institute of Nanoscience and Technology, Hanyang University, 222 Wangsimni-ro, Seongdong-gu, Seoul 133-791, Republic of Korea. Electronic address: justynj@hanyang.ac.kr.
authors.forename: Justyn
authors.initials: J
authors.lastname: Jaworski
mesh_terms: D004798:Enzymes / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry*; D013499:Surface Properties
chemical_list: D004798:Enzymes; D011506:Proteins
keywords: Covalent attachment; Formylglycine generating enzyme; Heterogeneous biocatalysis; Site-specific linkage; Surface modification; Un-natural side chains