title: Rosmarinic acid synthase is a new member of the superfamily of BAHD acyltransferases.
abstract: Purification of rosmarinic acid synthase (hydroxycinnamoyl-CoA:hydroxyphenyllactate hydroxycinnamoyltransferase) from suspension cells of Coleus blumei Benth. (Lamiaceae) by fractionated ammonium sulphate precipitation, hydrophobic interaction chromatography and two affinity chromatography steps led to the identification of peptide sequences, which enabled a PCR-based approach to isolate the full-length cDNA encoding this enzyme. The open reading frame of the cDNA had a length of 1290 base pairs encoding a protein of 430 amino acid residues with a molecular mass of 47,932 Da with typical characteristics of an acyltransferase of the BAHD superfamily. The cDNA was heterologously expressed in Escherichia coli. The enzyme displayed the activity of rosmarinic acid synthase using 4-coumaroyl- and caffeoyl-coenzyme A and 4-hydroxyphenyllactate as well as 3.4-dihydroxyphenyllactate as substrates. Shikimic acid and quinic acid were not able to serve as hydroxycinnamoyl acceptors. This therefore is the first report of the cDNA-cloning of a rosmarinic acid synthase.
authors.affiliation: Institut für Pharmazeutische Biologie, Philipps-Universität Marburg, Deutschhausstr. 17A, 35037, Marburg, Germany.
authors.forename: Anja
authors.initials: A
authors.lastname: Berger
authors.forename: Juliane
authors.initials: J
authors.lastname: Meinhard
authors.forename: Maike
authors.initials: M
authors.lastname: Petersen
mesh_terms: D000217:Acyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D001483:Base Sequence; D018076:DNA, Complementary; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction
chemical_list: D018076:DNA, Complementary; D000217:Acyltransferases