title: Molecular and biochemical characterization of the UDP-glucose: Anthocyanin 5-O-glucosyltransferase from Vitis amurensis.
abstract: Generally, red Vitis vinifera grapes only contain monoglucosidic anthocyanins, whereas most non-vinifera red grapes of the Vitis genus have both monoglucosidic and bis-glucosidic anthocyanins, the latter of which are believed to be more hydrophilic and more stable. Although previous studies have established the biosynthetic mechanism for formation of monoglucosidic anthocyanins, less attention has been paid to that of bis-glucosidic anthocyanins. In the present research, the full-length cDNA of UDP-glucose: anthocyanin 5-O-glucosyltransferase from Vitis amurensis Rupr. cv. 'Zuoshanyi' grape (Va5GT) was cloned. After acquisition and purification of recombinant Va5GT, its enzymatic parameters were systematically analyzed in vitro. Recombinant Va5GT used malvidin-3-O-glucoside as its optimum glycosidic acceptor when UDP-glucose was used as the glycosidic donor. Va5GT-GFP was found to be located in the cytoplasm by analyzing its subcellular localization with a laser-scanning confocal fluorescence microscope, and this result was coincident with its metabolic function of modifying anthocyanins in grape cells. Furthermore, the relationship between the transcriptional expression of Va5GT and the accumulation of anthocyanidin bis-glucosides during berry development suggested that Va5GT is a key enzyme in the biosynthesis of bis-glucosidic anthocyanins in V. amurensis grape berries.
authors.affiliation: Center for Viticulture & Enology, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China.
authors.forename: Fei
authors.initials: F
authors.lastname: He
authors.affiliation: Center for Viticulture & Enology, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China.
authors.forename: Wei-Kai
authors.initials: WK
authors.lastname: Chen
authors.affiliation: Center for Viticulture & Enology, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China.
authors.forename: Ke-Ji
authors.initials: KJ
authors.lastname: Yu
authors.affiliation: Center for Viticulture & Enology, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China.
authors.forename: Xiang-Nan
authors.initials: XN
authors.lastname: Ji
authors.affiliation: Center for Viticulture & Enology, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China.
authors.forename: Chang-Qing
authors.initials: CQ
authors.lastname: Duan
authors.affiliation: Center for Viticulture & Enology, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China; Institute of Food, Nutrition and Human Health, Massey University, Palmerston North 4442, New Zealand.
authors.forename: Malcolm J
authors.initials: MJ
authors.lastname: Reeves
authors.affiliation: Center for Viticulture & Enology, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China. Electronic address: jun_wang@cau.edu.cn.
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
mesh_terms: D000872:Anthocyanins / Q000378:metabolism; D003001:Cloning, Molecular; D003593:Cytoplasm / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005638:Fruit / Q000254:growth & development / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D005960:Glucosides / Q000378:metabolism; D005964:Glucosyltransferases / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence / Q000379:methods; D010802:Phylogeny; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity; D027843:Vitis / Q000235:genetics / Q000378:metabolism*
chemical_list: D000872:Anthocyanins; D005960:Glucosides; D011994:Recombinant Proteins; C458419:malvidin-3-glucoside; D005964:Glucosyltransferases; C118149:UDP-glucose anthocyanin 5-O-glucosyltransferase
keywords: Amur grape; Bis-glucosidic anthocyanin; Enzymology; Subcellular localization; UDP-glucose: anthocyanin 5-O-glucosyltransferase; Vitaceae; Vitis amurensis