title: Cytochrome P450 CYP71BE5 in grapevine (Vitis vinifera) catalyzes the formation of the spicy aroma compound (-)-rotundone.
abstract: (-)-Rotundone is a potent odorant molecule with a characteristic spicy aroma existing in various plants including grapevines (Vitis vinifera). It is considered to be a significant compound in wines and grapes because of its low sensory threshold and aroma properties. (-)-Rotundone was first identified in red wine made from the grape cultivar Syrah and here we report the identification of VvSTO2 as a α-guaiene 2-oxidase which can transform α-guaiene to (-)-rotundone in the grape cultivar Syrah. It is a cytochrome P450 (CYP) enzyme belonging to the CYP 71BE subfamily, which overlaps with the very large CYP71D family and, to the best of our knowledge, this is the first functional characterization of an enzyme from this family. VvSTO2 was expressed at a higher level in the Syrah grape exocarp (skin) in accord with the localization of (-)-rotundone accumulation in grape berries. α-Guaiene was also detected in the Syrah grape exocarp at an extremely high concentration. These findings suggest that (-)-rotundone accumulation is regulated by the VvSTO2 expression along with the availability of α-guaiene as a precursor. VvSTO2 expression during grape maturation was considerably higher in Syrah grape exocarp compared to Merlot grape exocarp, consistent with the patterns of α-guaiene and (-)-rotundone accumulation. On the basis of these findings, we propose that VvSTO2 may be a key enzyme in the biosynthesis of (-)-rotundone in grapevines by acting as a α-guaiene 2-oxidase.
authors.affiliation: Laboratory, New Product & Process Developments, Mercian Corporation, 4-9-1 Johnan, Fujisawa, Kanagawa 251-0057, Japan The Institute of Enology and Viticulture, University of Yamanashi, 1-13-1 Kitashin, Kofu, Yamanashi 400-0005, Japan.
authors.forename: Hideki
authors.initials: H
authors.lastname: Takase
authors.affiliation: Laboratory, New Product & Process Developments, Mercian Corporation, 4-9-1 Johnan, Fujisawa, Kanagawa 251-0057, Japan.
authors.forename: Kanako
authors.initials: K
authors.lastname: Sasaki
authors.affiliation: Interdisciplinary Graduate School of Medical and Engineering, University of Yamanashi, 4-4-37 Takeda, Kofu, Yamanashi 400-8510, Japan.
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Shinmori
authors.affiliation: Interdisciplinary Graduate School of Medical and Engineering, University of Yamanashi, 4-4-37 Takeda, Kofu, Yamanashi 400-8510, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Shinohara
authors.affiliation: Interdisciplinary Graduate School of Medical and Engineering, University of Yamanashi, 4-4-37 Takeda, Kofu, Yamanashi 400-8510, Japan.
authors.forename: Chihiro
authors.initials: C
authors.lastname: Mochizuki
authors.affiliation: Château Mercian, 1425-1 Shimoiwasaki, Katsunuma, Koshu, Yamanashi 409-1313, Japan.
authors.forename: Hironori
authors.initials: H
authors.lastname: Kobayashi
authors.affiliation: The Institute of Enology and Viticulture, University of Yamanashi, 1-13-1 Kitashin, Kofu, Yamanashi 400-0005, Japan.
authors.forename: Gen
authors.initials: G
authors.lastname: Ikoma
authors.affiliation: Château Mercian, 1425-1 Shimoiwasaki, Katsunuma, Koshu, Yamanashi 409-1313, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Saito
authors.affiliation: Château Mercian, 1425-1 Shimoiwasaki, Katsunuma, Koshu, Yamanashi 409-1313, Japan.
authors.forename: Hironori
authors.initials: H
authors.lastname: Matsuo
authors.affiliation: The Institute of Enology and Viticulture, University of Yamanashi, 1-13-1 Kitashin, Kofu, Yamanashi 400-0005, Japan suzukis@yamanashi.ac.jp.
authors.forename: Shunji
authors.initials: S
authors.lastname: Suzuki
authors.affiliation: Laboratory, New Product & Process Developments, Mercian Corporation, 4-9-1 Johnan, Fujisawa, Kanagawa 251-0057, Japan.
authors.forename: Ryoji
authors.initials: R
authors.lastname: Takata
mesh_terms: D000595:Amino Acid Sequence; D052176:Azulenes / Q000378:metabolism; D055162:Biocatalysis; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D057075:Enzyme Assays; D005638:Fruit / Q000201:enzymology / Q000235:genetics; D008401:Gas Chromatography-Mass Spectrometry; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D007700:Kinetics; D008969:Molecular Sequence Data; D009812:Odorants; D010802:Phylogeny; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic / Q000235:genetics; D016415:Sequence Alignment; D012717:Sesquiterpenes / Q000378:metabolism*; D045789:Sesquiterpenes, Guaiane / Q000378:metabolism; D013379:Substrate Specificity; D027843:Vitis / Q000201:enzymology* / Q000235:genetics
chemical_list: D052176:Azulenes; D010940:Plant Proteins; D012333:RNA, Messenger; D012717:Sesquiterpenes; D045789:Sesquiterpenes, Guaiane; C530290:rotundone; C019408:guaiene; D003577:Cytochrome P-450 Enzyme System
keywords: Cytochrome P450; Vitis vinifera; grapevine; guaiene; rotundone; sesquiterpene oxidase; wine.