Molecular Cloning, Expression and Characterisation of a Bacterial Myrosinase from Citrobacter Wye1

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dc.authoridCebeci, Fatma/0000-0003-4715-6689
dc.contributor.authorCebeci, Fatma
dc.contributor.authorMayer, Melinda J.
dc.contributor.authorRossiter, John T.
dc.contributor.authorMithen, Richard
dc.contributor.authorNarbad, Arjan
dc.date.accessioned2024-10-04T18:52:30Z
dc.date.available2024-10-04T18:52:30Z
dc.date.issued2022
dc.departmentBayburt Üniversitesien_US
dc.description.abstractGlucosinolates are plant natural products which on degradation by myrosinases give rise to the beneficial bioactive isothiocyanates. Recently, a myrosinase activity was detected in a Citrobacter strain isolated from soil. This enzyme was purified enabling its amino acid sequence and gene sequence (cmyr) to be determined. In order to study this myrosinase it was necessary to establish an expression system that would enable future work such as a structural determination of the protein to be carried out. The myrosinase gene was amplified, cloned and expressed in Escherichia coli with a 6XHis-tag. The heterologous expression of cmyr enabled relatively large amounts of myrosinase to be produced (3.4 mg cmyr/100 ml culture). Myrosinase activity was determined by mixing substrate and enzyme and determining glucose release. Optimum pH and temperature were determined to be pH 6.0 and 25 degrees C for the Ni-NTA purified protein. The kinetic parameters of the purified myrosinase were determined using sinigrin as a substrate. K-m and V-max were estimated as 0.18 mM and 0.033 mmol/min/mg respectively for sinigrin under optimum conditions and compared to other kinetic data for myrosinases. The substrate specificity of myrosinase was determined having the highest affinity for sinigrin followed by glucoiberin, progoitrin, glucoerucin, glucoraphanin and glucotropaeolin.en_US
dc.description.sponsorshipBiotechnological and Biological Sciences Research Council [BB/J004529/1, BB/J004545/1, BB/R012490/1, BB/ R012512/1]; Republic of Turkey Ministry of National Educationen_US
dc.description.sponsorshipQuadram Institute Bioscience is supported by the Biotechnological and Biological Sciences Research Council via Institute Strategic grants BB/J004529/1, BB/J004545/1, BB/R012490/1 and BB/ R012512/1. Fatma Cebeci was supported by a scholarship funded by the Republic of Turkey Ministry of National Education.en_US
dc.identifier.doi10.1007/s10930-021-10034-5
dc.identifier.endpage140en_US
dc.identifier.issn1572-3887
dc.identifier.issn1875-8355
dc.identifier.issue1en_US
dc.identifier.pmid35031980en_US
dc.identifier.scopus2-s2.0-85123125228en_US
dc.identifier.scopusqualityQ1en_US
dc.identifier.startpage131en_US
dc.identifier.urihttps://doi.org/10.1007/s10930-021-10034-5
dc.identifier.urihttp://hdl.handle.net/20.500.12403/3502
dc.identifier.volume41en_US
dc.identifier.wosWOS:000742585900001en_US
dc.identifier.wosqualityQ3en_US
dc.indekslendigikaynakWeb of Scienceen_US
dc.indekslendigikaynakScopusen_US
dc.indekslendigikaynakPubMeden_US
dc.language.isoenen_US
dc.publisherSpringeren_US
dc.relation.ispartofProtein Journalen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanıen_US
dc.rightsinfo:eu-repo/semantics/closedAccessen_US
dc.subjectCitrobacteren_US
dc.subjectMyrosinaseen_US
dc.subjectGlucosinolateen_US
dc.subjectBiotransformationen_US
dc.titleMolecular Cloning, Expression and Characterisation of a Bacterial Myrosinase from Citrobacter Wye1en_US
dc.typeArticleen_US

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