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dc.creatorBalaž, Ana Marija
dc.creatorStevanović, Jelena
dc.creatorOstafe, Raluca
dc.creatorBlazić, Marija
dc.creatorIlić Đurđić, Karla
dc.creatorFischer, Rainer
dc.creatorProdanović, Radivoje
dc.date.accessioned2020-08-03T08:03:37Z
dc.date.available2020-08-03T08:03:37Z
dc.date.issued2020
dc.identifier.issn1381-1991
dc.identifier.urihttp://cherry.chem.bg.ac.rs/handle/123456789/4011
dc.description.abstractCellobiose dehydrogenase (CDH, EC 1.1.99.18) from white rot fungi Phanerochaete chrysosporium can be used for constructing biosensors and biofuel cells, for bleaching cotton in textile industry, and recently, the enzyme has found an important application in biomedicine as an antimicrobial and antibiofilm agent. Stability and activity of the wild-type (wt) CDH and mutants at methionine residues in the presence of hydrogen peroxide were investigated. Saturation mutagenesis libraries were made at the only methionine in heme domain M65 and two methionines M685 and M738 in the flavin domain that were closest to the active site. After screening the libraries, three mutants with increased activity and stability in the presence of peroxide were found, M65F with 70% of residual activity after 6 h of incubation in 0.3 M hydrogen peroxide, M738S with 80% of residual activity and M685Y with over 90% of residual activity compared to wild-type CDH that retained 40% of original activity. Combined mutants showed no activity. The most stable mutant M685Y with 5.8 times increased half-life in the presence of peroxide showed also 2.5 times increased kcat for lactose compared to wtCDH and could be good candidate for applications in biofuel cells and biocatalysis for lactobionic acid production.
dc.relationinfo:eu-repo/grantAgreement/MESTD/Integrated and Interdisciplinary Research (IIR or III)/46010/RS//
dc.relationinfo:eu-repo/grantAgreement/MESTD/Basic Research (BR or ON)/172049/RS//
dc.relationinfo:eu-repo/grantAgreement/MESTD/Basic Research (BR or ON)/173017/RS//
dc.rightsrestrictedAccess
dc.sourceMolecular Diversity
dc.subjectProtein engineering
dc.subjectHydrogen peroxide
dc.subjectLibrary
dc.subjectMethionine
dc.subjectYeast
dc.titleSemi-rational design of cellobiose dehydrogenase for increased stability in the presence of peroxide
dc.typearticleen
dc.rights.licenseARR
dcterms.abstractПродановић, Радивоје; Стевановић, Јелена; Балаж, Aна Марија; Остафе, Ралуца; Илић Ђурђић, Карла; Фисцхер, Раинер; Блазић, Марија;
dcterms.publisherSpringer
dc.citation.volume24
dc.citation.issue3
dc.citation.spage593
dc.citation.epage601
dc.identifier.wos000547921900001
dc.identifier.doi10.1007/s11030-019-09965-0
dc.type.versionpublishedVersion
dc.identifier.scopus2-s2.0-85066612562


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