title: Cloning and functional expression in E. coli of a polyphenol oxidase transcript from Coreopsis grandiflora involved in aurone formation.
abstract: Polyphenol oxidases are involved in aurone biosynthesis but the gene responsible for 4-deoxyaurone formation in Asteraceae was so far unknown. Three novel full-length cDNA sequences were isolated from Coreopsis grandiflora with sizes of 1.80kb (cgAUS1) and 1.85kb (cgAUS2a, 2b), encoding for proteins of 68-69kDa, respectively. cgAUS1 is preferably expressed in young petals indicating a specific role in pigment formation. The 58.9kDa AUS1 holoproenzyme, was recombinantly expressed in E. coli and purified to homogeneity. The enzyme shows only diphenolase activity, catalyzing the conversion of chalcones to aurones and was characterized by SDS-PAGE and shot-gun type nanoUHPLC-ESI-MS/MS.
authors.affiliation: Universität Wien, Fakultät für Chemie, Institut für Biophysikalische Chemie, Althanstraße 14, 1090 Wien, Austria. Electronic address: cornelia.kaintz@univie.ac.at.
authors.forename: Cornelia
authors.initials: C
authors.lastname: Kaintz
authors.affiliation: Universität Wien, Fakultät für Chemie, Institut für Biophysikalische Chemie, Althanstraße 14, 1090 Wien, Austria. Electronic address: christian.molitor@univie.ac.at.
authors.forename: Christian
authors.initials: C
authors.lastname: Molitor
authors.affiliation: University of Technology Vienna, Institute of Chemical Engineering, Getreidemarkt 9, 1060 Vienna, Austria. Electronic address: jthill@mail.zserv.tuwien.ac.at.
authors.forename: Jana
authors.initials: J
authors.lastname: Thill
authors.affiliation: Universität Wien, Fakultät für Chemie, Institut für Biophysikalische Chemie, Althanstraße 14, 1090 Wien, Austria; University of Technology Vienna, Institute of Chemical Engineering, Getreidemarkt 9, 1060 Vienna, Austria. Electronic address: ioannis.kampatsikas@univie.ac.at.
authors.forename: Ioannis
authors.initials: I
authors.lastname: Kampatsikas
authors.affiliation: University of Vienna, Department of Analytical Chemistry, Währinger Straße 38, 1090 Vienna, Austria. Electronic address: claudia.michael@univie.ac.at.
authors.forename: Claudia
authors.initials: C
authors.lastname: Michael
authors.affiliation: University of Technology Vienna, Institute of Chemical Engineering, Getreidemarkt 9, 1060 Vienna, Austria. Electronic address: hhalb@mail.zserv.tuwien.ac.at.
authors.forename: Heidi
authors.initials: H
authors.lastname: Halbwirth
authors.affiliation: Universität Wien, Fakultät für Chemie, Institut für Biophysikalische Chemie, Althanstraße 14, 1090 Wien, Austria. Electronic address: annette.rompel@univie.ac.at.
authors.forename: Annette
authors.initials: A
authors.lastname: Rompel
mesh_terms: D000595:Amino Acid Sequence; D001572:Benzofurans / Q000378:metabolism*; D004156:Catechol Oxidase / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D003001:Cloning, Molecular; D031945:Coreopsis / Q000201:enzymology*; D004926:Escherichia coli; D035264:Flowers / Q000201:enzymology*; D015870:Gene Expression; D008969:Molecular Sequence Data; D009928:Organ Specificity; D010802:Phylogeny; D010858:Pigmentation; D010940:Plant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D017422:Sequence Analysis, DNA
chemical_list: D001572:Benzofurans; D010940:Plant Proteins; C486837:aurone; D004156:Catechol Oxidase
keywords: 4-Deoxyaurone; Aurone synthase; Coreopsis grandiflora; Heterologous expression; Polyphenol oxidase; Type-3 copper
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