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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.

Amine-functionalization of glycidyl methacrylate-containing emulsion-templated porous polymers and immobilization of proteinase K for biocatalysis Thumbnail


Authors

Scott D. Kimmins

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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Publisher Licence URL
http://creativecommons.org/licenses/by/4.0/

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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