@article {Willits:2004p8030,
	title = {Bio-fermentation of modified flavonoids: an example of in vivo diversification of secondary metabolites},
	journal = {Phytochemistry},
	volume = {65},
	number = {1},
	year = {2004},
	month = {Jan},
	pages = {31{\textendash}41},
	abstract = {A bio-fermentation technique was used for the in vivo diversification of flavonoid structures based on expression in Escherichia coli of six O-methyltransferases (OMTs) from Mentha x piperita and one O-glucosyltransferase (GT) each from Arabidopsis thaliana and Allium cepa. Enzymes were shown to be regio-specific in in vitro experiments and modified a broad range of flavonoid substrates at various positions. Using the flavonol quercetin as a model substrate, we show that the product spectrum produced with the in vivo approach is identical to that found in vitro. Additionally, using mixed cultures of E. coli expressing different classes of modifying genes (OMTs and GTs), the production of polymethylated flavonoid glucosides was observed. This report demonstrates the potential to increase the structural diversity of plant secondary metabolites using a multi-enzyme, bio-fermentation approach.},
	keywords = {Amino Acid Sequence, Arabidopsis, Cloning: Molecular, Consensus Sequence, Escherichia coli, Fermentation, Flavonoids, Glucosyltransferases, Mentha piperita, Methyltransferases, Molecular Sequence Data, Onions, Phylogeny, Recombinant Proteins, Sequence Alignment, Sequence Homology: Amino Acid, Stereoisomerism, Substrate Specificity},
	author = {Michael G Willits and Ma{\"{\i}t{\'e} Giovanni and Rogerio T N Prata and Catherine M Kramer and Vincenzo De Luca and John C Steffens and Gerson Graser}
}
@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 {Hirotani:2000p8040,
	title = {Cloning and expression of UDP-glucose: flavonoid 7-O-glucosyltransferase from hairy root cultures of Scutellaria baicalensis},
	journal = {Planta},
	volume = {210},
	number = {6},
	year = {2000},
	month = {May},
	pages = {1006{\textendash}13},
	abstract = {A cDNA encoding UDP-glucose: baicalein 7-O-glucosyltransferase (UBGT) was isolated from a cDNA library from hairy root cultures of Scutellaria baicalensis Georgi probed with a partial-length cDNA clone of a UDP-glucose: flavonoid 3-O-glucosyltransferase (UFGT) from grape (Vitis vinifera L.). The heterologous probe contained a glucosyltransferase consensus amino acid sequence which was also present in the Scutellaria cDNA clones. The complete nucleotide sequence of the 1688-bp cDNA insert was determined and the deduced amino acid sequences are presented. The nucleotide sequence analysis of UBGT revealed an open reading frame encoding a polypeptide of 476 amino acids with a calculated molecular mass of 53,094 Da. The reaction product for baicalein and UDP-glucose catalyzed by recombinant UBGT in Escherichia coli was identified as authentic baicalein 7-O-glucoside using high-performance liquid chromatography and proton nuclear magnetic resonance spectroscopy. The enzyme activities of recombinant UBGT expressed in E. coli were also detected towards flavonoids such as baicalein, wogonin, apigenin, scutellarein, 7,4{\textquoteright}-dihydroxyflavone and kaempferol, and phenolic compounds. The accumulation of UBGT mRNA in hairy roots was in response to wounding or salicylic acid treatments.},
	keywords = {Amino Acid Sequence, Blotting: Northern, Chromatography: High Pressure Liquid, Cloning: Molecular, Escherichia coli, Flavonoids, Glucosyltransferases, Lamiaceae, Magnetic Resonance Spectroscopy, Molecular Sequence Data, Phylogeny, Plant Roots, Recombinant Proteins},
	author = {M Hirotani and R Kuroda and H Suzuki and T Yoshikawa}
}
