title: Sorption-assisted surface conjugation: a way to stabilize laccase enzyme.
abstract: Enyzme immobilization on solid surfaces is one of the most relevant methods to improve enzyme activity and stability under harsh conditions over extended periods. A typically interesting application is the immobilization of laccases, multicopper enzymes oxidizing aromatic compounds, to solid surfaces in order to develop valuable tools for the elimination of micropollutants in wastewater. Laccase of the white-rot fungus Coriolopsis polyzona has been successfully immobilized on fumed silica nanoparticles using a novel method. It consists in the sorption of the enzyme to amino-modified silica nanoparticles and the subsequent covalent cross-linking using glutaraldehyde as a homobifunctional linker. The so-produced nanoparticulate material has been characterized by means of scanning electron microscopy and Brunauer-Emmett-Teller surface area analysis revealing modifications of the surface structure and area during the coupling procedure. Laccase immobilization on spherical nanoparticles produced according to the method of Stöber has been shown to be much less efficient than on fumed silica nanoparticles. Long-term stability assays revealed that the novel developed method allows a drastic stabilization of the enzyme. In real wastewater, 77% of the laccase activity remained on the nanoparticles over 1 month, whereas the activity of free laccase dropped to 2.5%. The activity loss on the nanoparticles resulted from partial inactivation of the immobilized enzymes and additional release into the surrounding solution with subsequent fast inactivation of the free enzymes, since almost no activity was found in the supernatants.
authors.affiliation: Institute for Ecopreneurship, School of Life Sciences, University of Applied Sciences Northwestern Switzerland, Muttenz, Switzerland.
authors.forename: Yannick-Serge
authors.initials: YS
authors.lastname: Zimmermann
authors.forename: Patrick
authors.initials: P
authors.lastname: Shahgaldian
authors.forename: Philippe F X
authors.initials: PF
authors.lastname: Corvini
authors.forename: Gregor
authors.initials: G
authors.lastname: Hommes
mesh_terms: D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000378:metabolism*; D042845:Laccase / Q000737:chemistry* / Q000378:metabolism*; D008854:Microscopy, Electron; D053758:Nanoparticles / Q000737:chemistry*; D012822:Silicon Dioxide / Q000737:chemistry*; D013997:Time Factors; D055454:Trametes / Q000201:enzymology*
chemical_list: D004800:Enzymes, Immobilized; D012822:Silicon Dioxide; D042845:Laccase