@article {Yamazaki:1999p8037,
	title = {Molecular cloning and biochemical characterization of a novel anthocyanin 5-O-glucosyltransferase by mRNA differential display for plant forms regarding anthocyanin},
	journal = {J Biol Chem},
	volume = {274},
	number = {11},
	year = {1999},
	month = {Mar},
	pages = {7405{\textendash}11},
	abstract = {UDP-glucose: anthocyanin 5-O-glucosyltransferase (5-GT) is responsible for the modification of anthocyanins to more stable molecules in complexes for co-pigmentation, supposedly resulting in a purple hue. The cDNA encoding 5-GT was isolated by a differential display applied to two different forms of anthocyanin production in Perilla frutescens var. crispa. Differential display was carried out for mRNA from the leaves of reddish-purple and green forms of P. frutescens, resulting in the isolation of five cDNA clones predominantly expressed in the red form. The cDNA encoded a polypeptide of 460 amino acids, exhibiting a low homology with the sequences of several glucosyltransferases including UDP-glucose: anthocyanidin 3-O-glucosyltransferase. By using this cDNA as the probe, we also isolated a homologous cDNA clone from a petal cDNA library of Verbena hybrida. To identify the biochemical function of the encoded proteins, these cDNAs were expressed in Saccharomyces cerevisiae cells. The recombinant proteins in the yeast extracts catalyzed the conversion of anthocyanidin 3-O-glucosides into the corresponding anthocyanidin 3,5-di-O-glucosides using UDP-glucose as a cofactor, indicating the identity of the cDNAs encoding 5-GT. Several biochemical properties (optimum pH, Km values, and sensitivity to inhibitors) were similar to those reported previously for 5-GTs. Southern blot analysis indicated the presence of two copies of 5-GT genes in the genome of both red and green forms of P. frutescens. The mRNA accumulation of the 5-GT gene was detected in the leaves of the red form but not in those of the green form and was induced by illumination of light, as observed for other structural genes for anthocyanin biosynthesis in P. frutescens.},
	keywords = {Amino Acid Sequence, Anthocyanins, Cloning: Molecular, DNA: Complementary, Glucosyltransferases, Lamiaceae, Molecular Sequence Data, Phylogeny, Recombinant Proteins, RNA: Messenger, Saccharomyces cerevisiae, Sequence Homology: Amino Acid},
	author = {M Yamazaki and Z Gong and M Fukuchi-Mizutani and Y Fukui and Y Tanaka and T Kusumi and K Saito}
}
@article {Suzuki:2001p8035,
	title = {Malonyl-CoA:anthocyanin 5-O-glucoside-6"{\textquoteright}-O-malonyltransferase from scarlet sage (Salvia splendens) flowers. Enzyme purification, gene cloning, expression, and characterization},
	journal = {J Biol Chem},
	volume = {276},
	number = {52},
	year = {2001},
	month = {Dec},
	pages = {49013{\textendash}9},
	abstract = {The 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"{\textquoteright}-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.},
	keywords = {Acyltransferases, Amino Acid Sequence, Anthocyanins, Molecular Sequence Data, Molecular Structure, Phylogeny, Pigmentation, Plant Structures, Salvia},
	doi = {10.1074/jbc.M108444200},
	author = {H Suzuki and T Nakayama and K Yonekura-Sakakibara and Y Fukui and N Nakamura and M Nakao and Y Tanaka and M A Yamaguchi and T Kusumi and T Nishino}
}
@article {Kitada:2001p8039,
	title = {Differential expression of two cytochrome P450s involved in the biosynthesis of flavones and anthocyanins in chemo-varietal forms of Perilla frutescens},
	journal = {Plant Cell Physiol},
	volume = {42},
	number = {12},
	year = {2001},
	month = {Dec},
	pages = {1338{\textendash}44},
	abstract = {In Perilla frutescens, there are two varietal forms of anthocyanin accumulation, i.e. red and green forms. The cDNA clones encoding flavone synthase II (FSII) and flavonoid 3{\textquoteright}-hydroxylase (F3{\textquoteright}H), two cytochrome P450s that are involved in the biosynthesis of flavones and anthocyanins, were isolated from P. frutescens. The FSII cDNA encoded a 57.1 kDa protein designated as CYP93B6, and the F3{\textquoteright}H cDNA encoded 57.5 kDa protein designated as CYP75B4. Recombinant CYP93B6 expressed in yeast converted flavanones to flavones with K(m) values of 8.8-11.9 microM. Recombinant CYP75B4 catalyzed 3{\textquoteright}-hydroxylation of flavanones to the corresponding compounds with K(m) values of 18-20 microM. The CYP93B6 transcript accumulated to an equal level in leaves of both red and green forms of P. frutescens, in agreement with the accumulation pattern of flavones in the leaves. However, the CYP75B4 transcript was predominantly expressed in the red form of P. frutescens, and its expression was induced by light in conjunction with other transcripts of biosynthetic enzymes of anthocyanin. These results indicate that gene expression of a set of anthocyanin biosynthetic enzymes including F3{\textquoteright}H is regulated coordinately only in the red form of P. frutescens but not in the green form, whilst FSII gene expression is controlled in a similar manner in red and green forms of P. frutescens.},
	keywords = {Anthocyanins, Apigenin, Cytochrome P-450 Enzyme System, Flavonoids, Gene Expression Regulation: Enzymologic, Gene Expression Regulation: Plant, Kinetics, Lamiaceae, Luteolin, Mixed Function Oxygenases, Phylogeny, Plant Structures},
	author = {C Kitada and Z Gong and Y Tanaka and M Yamazaki and K Saito}
}
