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A continuous spectrophotometric assay for determination of the aureusidin synthase activity of tyrosinase

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dc.contributor.authorJiménez-Atiénzar, Mercedes
dc.contributor.authorCabanes Cos, Juana
dc.contributor.authorEscribano Cebrián, Josefa
dc.contributor.authorGandía Herrero, Fernando
dc.contributor.authorPérez Gilabert, Manuela
dc.contributor.authorGarcía Carmona, Francisco
dc.contributor.departmentDepartamento de Bioquímica y Biología Molecular "A"es
dc.date.accessioned2025-07-18T10:53:51Z
dc.date.available2025-07-18T10:53:51Z
dc.date.issued2010-04-13
dc.description© 2009 John Wiley & Sons, Ltd. This document is the Published version of a Published Work that appeared in final form in Phytochemical Analysis. To access the final edited and published work see https://doi.org/10.1002/pca.1197es
dc.description.abstractIntroduction – Aurones (aureusidin glycosides) are plant flavonoids that provide yellow colour to the flowers of some orna-mental plants. In this study we analyse the capacity of tyrosinase to catalyse the synthesis of aureusidin by tyrosinase fromthe chalcone THC (2′,4′,6′,4–tetrahydroxychalcone).Objective – To develop a simple continuous spectrophotometric assay for the analysis of the spectrophotometric and kineticcharacteristics of THC oxidation by tyrosinase.Methodology – THC oxidation was routinely assayed by measuring the increase in absorbance at 415 nm vs. reaction time.Results – According to the mechanism proposed for tyrosinase, the enzymatic reaction involves the o-hydroxylation of themonophenol THC to the o-diphenol (PHC, 2′,4′,6′,3,4 – pentahydroxychalcone), which is then oxidised to the correspondingo-quinone in a second enzymatic step. This product is highly unstable and thus undergoes a series of fast chemical reactionsto produce aureusidin. In these experimental conditions, the optimum pH for THC oxidation is 4.5. The progress curvesobtained for THC oxidation showed the appearance of a lag period. The following kinetic parameters were also determined:Km = 0.12 mM , Vm = 13 mM /min, Vm/Km = 0.11/min.Conclusion – This method has made it possible to analyse the spectrophotometric and kinetic characteristics of THC by tyro-sinase. This procedure has the advantages of a short analysis time, straightforward measurement techniques and repro-ducibility. In addition, it also allows the study of tyrosinase inhibitors, such as tropolone.es
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dc.format.extent6es
dc.identifier.citationPhytochem. Anal. 2010, 21, 273–278
dc.identifier.doihttps://doi.org/10.1002/pca.1197
dc.identifier.issnPrint: 0958-0344
dc.identifier.issnElectronic: 1099-1565
dc.identifier.urihttp://hdl.handle.net/10201/157533
dc.languageenges
dc.publisherWiley
dc.relationThis work was supported by a grant from the DGI (Spain), ProyectoAGL2007-65907 and by “Programa de Ayudas a Grupos de Excelencia de la Región de Murcia”, Plan Regional de Ciencia y Tecnología 2007/2010 (Fundación Séneca, Agencia de Ciencia y Tecnología de la Región de Murcia).es
dc.relation.publisherversionhttps://analyticalsciencejournals.onlinelibrary.wiley.com/doi/10.1002/pca.1197
dc.rights.accessRightsinfo:eu-repo/semantics/restrictedAccess
dc.subjectAuroneses
dc.subjectAuresidin synthasees
dc.subjectChalconees
dc.subjectPolyphenol oxidasees
dc.subjectTyrosinasees
dc.subject.otherCDU::5 - Ciencias puras y naturales::57 - Biología::577 - Bioquímica. Biología molecular. Biofísicaes
dc.titleA continuous spectrophotometric assay for determination of the aureusidin synthase activity of tyrosinasees
dc.typeinfo:eu-repo/semantics/articlees
dspace.entity.typePublicationes
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relation.isAuthorOfPublication.latestForDiscovery14e0a51b-b9d3-49d0-bc89-ca3ba64290dc
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