Issue 29, 2015

Enzymatic multi-functionalization of microparticles under aqueous neutral conditions


The immobilization of proteins and enzymes on microparticles enables innovative biofunctionalization and facilitates the recycling of, for example, catalysts in a biocatalytic process. When enzymes are used for the immobilization of proteins to material surfaces, site-specificity can be achieved, and the immobilization reaction can proceed under mild reaction conditions in an aqueous environment. By selectively oxidizing the exposed tyrosyl side chains of protein to reactive o-quinones using molecular oxygen as electron acceptor, tyrosinase promotes the formation of new covalent bonds via 1,4-addition of nucleophilic moieties to the o-quinones. The introduction of tyrosinase-susceptible tyrosine residues (Y-tag) to a protein by genetic engineering can thus enable site-specific crosslinking and site-specific protein immobilization. In this study, several variants of the fluorescent protein GFPuv were produced in order to investigate the tyrosinase-mediated crosslinking and immobilization reaction. The Y-tag was a target for rapid protein–protein crosslinking by tyrosinase catalysis. Moreover, low concentrations of tyrosinase were sufficient to obtain detectable fluorescent microparticles using the Y-tagged GFPuv variants, e.g. 0.4 μM (50 μg ml−1) protein concentrations. Eventually, we showed that this enzyme-based technology allows for the multiple functionalization of microparticles using different fluorescent proteins.

Graphical abstract: Enzymatic multi-functionalization of microparticles under aqueous neutral conditions

Supplementary files

Article information

Article type
12 Jan 2015
13 Feb 2015
First published
16 Feb 2015
This article is Open Access
Creative Commons BY license

RSC Adv., 2015,5, 22319-22325

Author version available

Enzymatic multi-functionalization of microparticles under aqueous neutral conditions

G. Faccio, S. Senkalla, L. Thöny-Meyer and M. Richter, RSC Adv., 2015, 5, 22319 DOI: 10.1039/C5RA00669D

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