Enzyme directed formation of un-natural side-chains for covalent surface attachment of proteins
- Title
- Enzyme directed formation of un-natural side-chains for covalent surface attachment of proteins
- Author
- Justyn Wayne Jaworski
- Keywords
- Un-natural side chains; Formylglycine generating enzyme; Heterogeneous biocatalysis; Surface modification; Covalent attachment; Site-specific linkage
- Issue Date
- 2014-06
- Publisher
- ELSEVIER SCIENCE BV, PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS
- Citation
- COLLOIDS AND SURFACES B-BIOINTERFACES, 권: 122, 페이지: 846-850
- 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. (c) 2014 Elsevier B.V. All rights reserved.
- URI
- http://www.sciencedirect.com/science/article/pii/S0927776514004329?via%3Dihubhttp://hdl.handle.net/20.500.11754/46756
- ISSN
- 0927-7765; 1873-4367
- DOI
- 10.1016/j.colsurfb.2014.08.010
- Appears in Collections:
- COLLEGE OF ENGINEERING[S](공과대학) > CHEMICAL ENGINEERING(화학공학과) > Articles
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