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dc.creatorOstafe, Raluca
dc.creatorProdanović, Radivoje
dc.creatorNazor, Jovana
dc.creatorFischer, Rainer
dc.date.accessioned2018-11-22T00:27:22Z
dc.date.available2018-11-22T00:27:22Z
dc.date.issued2014
dc.identifier.issn1074-5521
dc.identifier.urihttps://cherry.chem.bg.ac.rs/handle/123456789/1519
dc.description.abstractGlucose oxidase (GOx) is used in many industrial processes that could benefit from improved versions of the enzyme. Some improvements like higher activity under physiological conditions and thermal stability could be useful for GOx applications in biosensors and biofuel cells. Directed evolution is one of the currently available methods to engineer improved GOx variants. Here, we describe an ultra-high-throughput screening system for sorting the best enzyme variants generated by directed evolution that incorporates several methodological refinements: flow cytometry, in vitro compartmentalization, yeast surface display, fluorescent labeling of the expressed enzyme, delivery of glucose substrate to the reaction mixture through the oil phase, and covalent labeling of the cells with fluorescein-tyramide. The method enables quantitative screening of gene libraries to identify clones with improved activity and it also allows cells to be selected based not only on the overall activity but also on the specific activity of the enzyme.en
dc.publisherCell Press, Cambridge
dc.rightsopenAccess
dc.rights.urihttps://creativecommons.org/licenses/by-nc/4.0/
dc.sourceChemistry and Biology
dc.titleUltra-High-Throughput Screening Method for the Directed Evolution of Glucose Oxidaseen
dc.typearticle
dc.rights.licenseBY-NC
dcterms.abstractНазор, Јована; Остафе, Ралуца; Продановић, Радивоје; Фисцхер, Раинер;
dc.citation.volume21
dc.citation.issue3
dc.citation.spage414
dc.citation.epage421
dc.identifier.wos000333407000014
dc.identifier.doi10.1016/j.chembiol.2014.01.010
dc.citation.other21(3): 414-421
dc.citation.rankM21
dc.identifier.pmid24613019
dc.type.versionpublishedVersion
dc.identifier.scopus2-s2.0-84897065790
dc.identifier.fulltexthttps://cherry.chem.bg.ac.rs/bitstream/id/8630/1517.pdf


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