02941nas a2200349   4500008004100000245016900041210006900210260000800279300001400287490000800301520176300309653002402072653001702096653002302113653002302136653002502159653001402184653002802198653001402226653002502240653001902265653002902284653003402313100001602347700001202363700002402375700001302399700001402412700001402426700001302440856013802453       1999                        eng d00aMolecular cloning and biochemical characterization of a novel anthocyanin 5-O-glucosyltransferase by mRNA differential display for plant forms regarding anthocyanin0 aMolecular cloning and biochemical characterization of a novel an  cMar  a7405–110 v2743 aUDP-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.10aAmino Acid Sequence10aAnthocyanins10aCloning: Molecular10aDNA: Complementary10aGlucosyltransferases10aLamiaceae10aMolecular Sequence Data10aPhylogeny10aRecombinant Proteins10aRNA: Messenger10aSaccharomyces cerevisiae10aSequence Homology: Amino Acid1 aYamazaki, M1 aGong, Z1 aFukuchi-Mizutani, M1 aFukui, Y1 aTanaka, Y1 aKusumi, T1 aSaito, K  uhttps://lamiaceae.myspecies.info/content/molecular-cloning-and-biochemical-characterization-novel-anthocyanin-5-o-glucosyltransferase02472nas a2200325   4500008004100000245015600041210006900197260000800266300001400274490000700288520137700295653001701672653001301689653003501702653001501737653004401752653003801796653001301834653001401847653001301861653003001874653001401904653002101918100001401939700001201953700001401965700001601979700001301995856013802008       2001                        eng d00aDifferential expression of two cytochrome P450s involved in the biosynthesis of flavones and anthocyanins in chemo-varietal forms of Perilla frutescens0 aDifferential expression of two cytochrome P450s involved in the   cDec  a1338–440 v423 aIn 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'-hydroxylase (F3'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'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'-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'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.10aAnthocyanins10aApigenin10aCytochrome P-450 Enzyme System10aFlavonoids10aGene Expression Regulation: Enzymologic10aGene Expression Regulation: Plant10aKinetics10aLamiaceae10aLuteolin10aMixed Function Oxygenases10aPhylogeny10aPlant Structures1 aKitada, C1 aGong, Z1 aTanaka, Y1 aYamazaki, M1 aSaito, K  uhttps://lamiaceae.myspecies.info/content/differential-expression-two-cytochrome-p450s-involved-biosynthesis-flavones-and-anthocyanins