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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 | Schools: | School of Materials Science & Engineering | 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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