Scott D. Kimmins
Amine-functionalization of glycidyl methacrylate-containing emulsion-templated porous polymers and immobilization of proteinase K for biocatalysis
Kimmins, Scott D.; Wyman, Paul; Cameron, Neil R.
Authors
Paul Wyman
Neil R. Cameron
Abstract
Glycidyl methacrylate (GMA) emulsion-templated porous polymers (polyHIPEs) were prepared by thermal and photopolymerisation and derivatised with morpholine, tris(2-aminoethylamine) and a bisamino-PEG homobifunctional molecule. The extent of the functionalization reactions was investigated by a range of qualitative and quantitative techniques (FTIR, CHN analysis, titration, XPS, HR-MAS NMR spectroscopy, ninhydrin assay and Fmoc number determination) and was found to be excellent for small molecule amines (up to 89% conversion) but low for the reaction with PEG (2% conversion). This was ascribed to the high exclusion volume of the PEG chains in solution. Proteinase K (Pro K) was subsequently immobilized covalently onto the GMA polyHIPE material, both directly via reaction with surface epoxy groups and indirectly by activation of the pendent amine groups of PEGylated polyHIPE with glutaraldehyde then reductive amination with the enzyme. The activity of the supported enzymes was determined by a continuous electrochemical assay involving the hydrolysis of N-acetyl-l-tyrosine ethyl ester. The directly immobilized Pro K was found to have an activity of only 3.6 U/g whereas the activity of the enzyme immobilized via the PEG linker was much higher (up to 78 U/g).
Citation
Kimmins, S. D., Wyman, P., & Cameron, N. R. (2014). Amine-functionalization of glycidyl methacrylate-containing emulsion-templated porous polymers and immobilization of proteinase K for biocatalysis. Polymer, 55(1), 416-425. https://doi.org/10.1016/j.polymer.2013.09.019
Journal Article Type | Article |
---|---|
Publication Date | Jan 14, 2014 |
Deposit Date | Aug 2, 2013 |
Publicly Available Date | Feb 11, 2014 |
Journal | Polymer |
Print ISSN | 0032-3861 |
Publisher | Elsevier |
Peer Reviewed | Peer Reviewed |
Volume | 55 |
Issue | 1 |
Pages | 416-425 |
DOI | https://doi.org/10.1016/j.polymer.2013.09.019 |
Keywords | Porous polymers, Emulsions, Chemical functionalization. |
Public URL | https://durham-repository.worktribe.com/output/1472167 |
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Copyright Statement
This is an open-access article distributed under the terms of the Creative
Commons Attribution License, which permits unrestricted use, distribution, and
reproduction in any medium, provided the original author and source are credited.
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