03120nas 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