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dc.creatorBlažić, Marija
dc.creatorBalaž, Ana Marija
dc.creatorProdanović, Olivera
dc.creatorPopović, Nikolina
dc.creatorOstafe, Raluca
dc.creatorFischer, Rainer
dc.creatorProdanović, Radivoje
dc.date.accessioned2019-05-18T09:53:55Z
dc.date.available2020-05-14
dc.date.issued2019
dc.identifier.issn2076-3417
dc.identifier.urihttp://cer.ihtm.bg.ac.rs/handle/123456789/2875
dc.description.abstractCellobiose dehydrogenase (CDH) from Phanerochaete chrysosporium can be used in lactobionic acid production, biosensor for lactose, biofuel cells, lignocellulose degradation, and wound-healing applications. To make it a better biocatalyst, CDH with higher activity in an immobilized form is desirable. For this purpose, CDH was expressed for the first time on the surface of S. cerevisiae EBY100 cells in an active form as a triple mutant tmCDH (D20N, A64T, V592M) and evolved further for higher activity using resazurin-based fluorescent assay. In order to decrease blank reaction of resazurin with yeast cells and to have linear correlation between enzyme activity on the cell surface and fluorescence signal, the assay was optimized with respect to resazurin concentration (0.1 mM), substrate concentration (10mMlactose and 0.08mMcellobiose), and pH (6.0). Using optimized assay an error prone PCR gene library of tmCDH was screened. Two mutants with 5 (H5) and 7 mutations (H9) were found having two times higher activity than the parent tmCDH enzyme that already had improved activity compared to wild type CDH whose activity could not be detected on the surface of yeast cells.sr
dc.language.isoensr
dc.publisherBasel : MDPIsr
dc.relationinfo:eu-repo/grantAgreement/MESTD/Basic Research (BR or ON)/173017/RS//sr
dc.rightsopenAccesssr
dc.sourceApplied Sciencessr
dc.subjectcellobiose dehydrogenasesr
dc.subjectresazurinsr
dc.subjectfluorescent assaysr
dc.subjectflow cytometrysr
dc.subjectyeast surface displaysr
dc.titleDirected Evolution of Cellobiose Dehydrogenase on the Surface of Yeast Cells Using Resazurin-Based Fluorescent Assayen
dc.typearticlesr
dc.rights.licenseBYsr
dcterms.abstractПродановић, Оливера; Балаж, Aна Марија; Блажић, Марија; Поповић, Николина; Остафе, Ралуца; Продановић, Радивоје; Фисцхер, Раинер;
dc.citation.volume9
dc.citation.issue7
dc.citation.spage1413
dc.citation.rankM22~
dc.identifier.doi10.3390/app9071413
dc.identifier.fulltexthttp://cer.ihtm.bg.ac.rs//bitstream/id/6910/applsci-09-01413.pdf
dc.identifier.scopus2-s2.0-85064083412
dc.identifier.wos000466547500148
dc.type.versionpublishedVersionsr


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