Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/86561
Title: Protein engineering for covalent immobilization and enhanced stability through incorporation of multiple noncanonical amino acids
Authors: Deepankumar, Kanagavel
Prabhu, Nadarajan Saravanan
Kim, June-Hyung
Yun, Hyungdon
Keywords: Multiple noncanonical amino acids
Fluoroproline
Issue Date: 2017
Source: Deepankumar, K., Prabhu, N. S., Kim, J.-H., & Yun, H. (2017). Protein engineering for covalent immobilization and enhanced stability through incorporation of multiple noncanonical amino acids. Biotechnology and Bioprocess Engineering, 22(3), 248-255.
Series/Report no.: Biotechnology and Bioprocess Engineering
Abstract: In this study, we demonstrate the application of multiple functional properties of proteins generated through coupling of residue-specific and site-specific incorporation method. With green fluorescent protein (GFP) as a model protein, we constructed multifunctional GFP through sitespecific incorporation of L-3,4-dihydroxyphenylalanine (DOPA) and residue-specific incorporation of (2S, 4S)-4- fluoroproline (4S-FP) or L-homopropargylglycine (hpg). Fluorescence analysis revealed a conjugation efficiency of approximately 20% for conjugation of DOPA-containing variants GFPdopa, GFPdp[4S-FP], and GFPdphpg onto chitosan. While incorporation of 4S-FP improved protein folding and stability, hpg incorporation into GFP allowed conjugation with fluorescent dye/polyethylene glycol (PEG). In addition, the modification of GFPhpg and GFPdphpg with PEG through Cu(I)-catalyzed click reaction increased protein thermal stability by about two-fold of the wild-type GFP.
URI: https://hdl.handle.net/10356/86561
http://hdl.handle.net/10220/44074
ISSN: 1226-8372
DOI: 10.1007/s12257-017-0127-y
Rights: © 2017 The Korean Society for Biotechnology and Bioengineering and Springer
Fulltext Permission: none
Fulltext Availability: No Fulltext
Appears in Collections:MSE Journal Articles

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