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dc.creatorBlažić, Marija
dc.creatorBalaž, Ana Marija
dc.creatorTadić, Vojin
dc.creatorDraganić, Bojana
dc.creatorOstafe, Raluca
dc.creatorFischer, Rainer
dc.creatorProdanović, Radivoje
dc.date.accessioned2019-04-18T07:48:26Z
dc.date.available2021-03-25
dc.date.issued2019
dc.identifier.issn1369-703X
dc.identifier.urihttps://cer.ihtm.bg.ac.rs/handle/123456789/2655
dc.description.abstractCellobiose dehydrogenase (CDH) can be used in industry for lactobionic acid production, as a part of biosensors for disaccharides and in wound healing. In fungi it is involved in lignocellulose degradation. CDH gene from Phanerochaete chrysosporium has been cloned in pYES2 plasmid for extracellular expression and protein engineering in yeast Saccharomyces cerevisiae InvSC1 for the first time. A CDH gene library was generated using error-prone PCR and screened by spectrophotometric enzymatic assay based on 2,6-dichloroindophenol reduction detection in microtiter plates. Several mutants with increased activity and specificity towards lactose and cellobiose were found, purified and characterized in detail. Recombinant CDH enzymes showed a broad molecular weight between 120 and 150 KDa due to hyperglycosylation and the best S137N mutant showed 2.2 times increased kcat and 1.5 and 2 times increased specificity constant for lactose and cellobiose compared to the wild type enzyme. pH optimum of mutants was not changed while thermostability of selected mutants improved and S137N mutant retained 30% of it’s original activity after 15 minutes at 70oC compared to 10% of activity that the wild type enzyme retained. Mutants M65S and S137N showed also 1.6 and 1.5 times increased productivity of hydrogen peroxide in the presence of 30mM lactose compared to the wild type.en
dc.language.isoensr
dc.publisherElseviersr
dc.relationinfo:eu-repo/grantAgreement/MESTD/Basic Research (BR or ON)/172049/RS//sr
dc.relationinfo:eu-repo/grantAgreement/MESTD/Basic Research (BR or ON)/173017/RS//sr
dc.rightsembargoedAccesssr
dc.rights.urihttps://creativecommons.org/licenses/by-nc-nd/4.0/
dc.sourceBiochemical Engineering Journalsr
dc.subjectcellobiose dehydrogenasesr
dc.subjectdirected evolutionsr
dc.subjectlactosesr
dc.subjectSaccharomyces cerevisiaesr
dc.titleProtein engineering of cellobiose dehydrogenase from Phanerochaete chrysosporium in yeast Saccharomyces cerevisiae InvSc1 for increased activity and stabilityen
dc.typearticlesr
dc.rights.licenseBY-NC-NDsr
dcterms.abstractБлажић, Марија; Тадић, Војин; Драганић, Бојана; Остафе, Ралуца; Продановић, Радивоје; Фисцхер, Раинер; Балаж, Aна Марија;
dc.rights.holderElseviersr
dc.citation.volume146
dc.citation.spage179
dc.citation.epage185
dc.citation.rankM21
dc.description.otherThis is the peer-reviewed version of the article: [https://doi.org/10.1016/j.bej.2019.03.025]en
dc.description.other[http://cer.ihtm.bg.ac.rs/handle/123456789/2656]
dc.identifier.doi10.1016/j.bej.2019.03.025
dc.identifier.fulltexthttps://cer.ihtm.bg.ac.rs/bitstream/id/6278/bitstream_6278.pdf
dc.identifier.scopus2-s2.0-85063648705
dc.identifier.wos000466999900020
dc.type.versionacceptedVersionsr


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