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Cloning and functional expression in E. coli of a polyphenol oxidase transcript from Coreopsis grandiflora involved in aurone formation
Author(s) -
Kaintz Cornelia,
Molitor Christian,
Thill Jana,
Kampatsikas Ioannis,
Michael Claudia,
Halbwirth Heidi,
Rompel Annette
Publication year - 2014
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2014.07.034
Subject(s) - polyphenol oxidase , complementary dna , polyphenol , cloning (programming) , biochemistry , asteraceae , petal , chemistry , enzyme , biosynthesis , biology , gene , microbiology and biotechnology , botany , peroxidase , antioxidant , computer science , programming language
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 1.80 kb ( cgAUS1 ) and 1.85 kb ( cgAUS2a , 2b ), encoding for proteins of 68–69 kDa, respectively. cgAUS1 is preferably expressed in young petals indicating a specific role in pigment formation. The 58.9 kDa 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.