01879nas a2200241   4500008004100000245008900041210006900130260000800199300001400207490000800221520107600229653002101305653002401326653001801350653002301368653002801391653001401419653003001433100001701463700002201480700002001502856011501522       2006                        eng d00aRosmarinic acid synthase is a new member of the superfamily of BAHD acyltransferases0 aRosmarinic acid synthase is a new member of the superfamily of B  cNov  a1503–100 v2243 aPurification 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.10aAcyltransferases10aAmino Acid Sequence10aBase Sequence10aDNA: Complementary10aMolecular Sequence Data10aPhylogeny10aPolymerase Chain Reaction1 aBerger, Anja1 aMeinhard, Juliane1 aPetersen, Maike  uhttps://lamiaceae.myspecies.info/content/rosmarinic-acid-synthase-new-member-superfamily-bahd-acyltransferases03120nas a2200349   4500008004100000245018100041210006900222260000800291300001400299490000800313520197500321653002102296653002402317653001702341653002802358653002402386653001402410653001702424653002102441653001102462100001402473700001602487700002702503700001302530700001602543700001302559700001402572700002002586700001402606700001502620856013502635       2001                        eng d00aMalonyl-CoA:anthocyanin 5-O-glucoside-6"'-O-malonyltransferase from scarlet sage (Salvia splendens) flowers. Enzyme purification, gene cloning, expression, and characterization0 aMalonylCoAanthocyanin 5Oglucoside6Omalonyltransferase from scarl  cDec  a49013–90 v2763 aThe orange to blue coloration of flowers in nature is, in most cases, provided by anthocyanins, a class of plant flavonoids, many of which are modified by malonyl group(s). However, the identity of the enzyme catalyzing the malonylation reaction remains to be established. Here, we describe for the first time the purification, characterization, and cDNA cloning of an anthocyanin malonyltransferase from scarlet sage (Salvia splendens) flowers. The purified enzyme (termed Ss5MaT1) was a monomeric 50-kDa protein catalyzing the regiospecific transfer of the malonyl group from malonyl-CoA to the 6"'-hydroxyl group of the 5-glucosyl moiety of anthocyanins. Ss5MaT1 showed a k(cat) value of 7.8 s(-1) at 30 degrees C and pH 7.0 for the malonylation of bisdemalonylsalvianin (pelargonidin 3-(6"-O-caffeyl-beta-glucopyranoside)-5-beta-glucopyranoside) and K(m) values of 101 microm and 57 microm for bisdemalonylsalvianin and malonyl-CoA, respectively. p-Coumaric acid, which mimics an aromatic acyl group linked to the 3-glucosidic moiety of an anthocyanin substrate, was a competitive inhibitor with respect to the substrate. This strongly suggests that the presence of an aromatic acyl group at the 3-glucosidic moiety of anthocyanin is important for substrate recognition by the enzyme. On the basis of the partial amino acid sequences of the purified enzyme, we isolated a cDNA encoding Ss5MaT1. Ss5MaT1 consisted of 462 amino acids and shared motifs that are commonly found among members of a versatile plant acyltransferase family, which was recently shown to include numerous homologs of unknown biochemical functions. Northern blot analysis revealed that the transcripts of Ss5MaT1 were detected in petals, sepals, bracts, and red stems, in accordance with the pigment accumulation patterns. Phylogenetic analysis suggests that the aliphatic and aromatic acylations of anthocyanins are generally catalyzed by subfamily members of the plant acyltransferase family.10aAcyltransferases10aAmino Acid Sequence10aAnthocyanins10aMolecular Sequence Data10aMolecular Structure10aPhylogeny10aPigmentation10aPlant Structures10aSalvia1 aSuzuki, H1 aNakayama, T1 aYonekura-Sakakibara, K1 aFukui, Y1 aNakamura, N1 aNakao, M1 aTanaka, Y1 aYamaguchi, M, A1 aKusumi, T1 aNishino, T  uhttps://lamiaceae.myspecies.info/content/malonyl-coaanthocyanin-5-o-glucoside-6-o-malonyltransferase-scarlet-sage-salvia-splendens