title: Metabolic engineering for the production of prenylated polyphenols in transgenic legume plants using bacterial and plant prenyltransferases.
abstract: Prenylated polyphenols are secondary metabolites beneficial for human health because of their various biological activities. Metabolic engineering was performed using Streptomyces and Sophora flavescens prenyltransferase genes to produce prenylated polyphenols in transgenic legume plants. Three Streptomyces genes, NphB, SCO7190, and NovQ, whose gene products have broad substrate specificity, were overexpressed in a model legume, Lotus japonicus, in the cytosol, plastids or mitochondria with modification to induce the protein localization. Two plant genes, N8DT and G6DT, from Sophora flavescens whose gene products show narrow substrate specificity were also overexpressed in Lotus japonicus. Prenylated polyphenols were undetectable in these plants; however, supplementation of a flavonoid substrate resulted in the production of prenylated polyphenols such as 7-O-geranylgenistein, 6-dimethylallylnaringenin, 6-dimethylallylgenistein, 8-dimethylallynaringenin, and 6-dimethylallylgenistein in transgenic plants. Although transformants with the native NovQ did not produce prenylated polyphenols, modification of its codon usage led to the production of 6-dimethylallylnaringenin and 6-dimethylallylgenistein in transformants following naringenin supplementation. Prenylated polyphenols were not produced in mitochondrial-targeted transformants even under substrate feeding. SCO7190 was also expressed in soybean, and dimethylallylapigenin and dimethylallyldaidzein were produced by supplementing naringenin. This study demonstrated the potential for the production of novel prenylated polyphenols in transgenic plants. In particular, the enzymatic properties of prenyltransferases seemed to be altered in transgenic plants in a host species-dependent manner.
authors.affiliation: Laboratory of Plant Gene Expression, Research Institute for Sustainable Humanosphere, Kyoto University, Uji 611-0011, Japan.
authors.forename: Akifumi
authors.initials: A
authors.lastname: Sugiyama
authors.forename: Philip J
authors.initials: PJ
authors.lastname: Linley
authors.forename: Kanako
authors.initials: K
authors.lastname: Sasaki
authors.forename: Takuto
authors.initials: T
authors.lastname: Kumano
authors.forename: Hideaki
authors.initials: H
authors.lastname: Yamamoto
authors.forename: Nobukazu
authors.initials: N
authors.lastname: Shitan
authors.forename: Kazuaki
authors.initials: K
authors.lastname: Ohara
authors.forename: Kojiro
authors.initials: K
authors.lastname: Takanashi
authors.forename: Emiko
authors.initials: E
authors.lastname: Harada
authors.forename: Hisakazu
authors.initials: H
authors.lastname: Hasegawa
authors.forename: Teruhiko
authors.initials: T
authors.lastname: Terakawa
authors.forename: Tomohisa
authors.initials: T
authors.lastname: Kuzuyama
authors.forename: Kazufumi
authors.initials: K
authors.lastname: Yazaki
mesh_terms: D004122:Dimethylallyltranstransferase / Q000235:genetics / Q000378:metabolism*; D044950:Flavanones / Q000008:administration & dosage; D000070116:Lotus / Q000201:enzymology* / Q000235:genetics; D060847:Metabolic Engineering / Q000379:methods*; D030821:Plants, Genetically Modified / Q000201:enzymology* / Q000235:genetics; D059808:Polyphenols / Q000096:biosynthesis*; D054876:Prenylation / Q000235:genetics; D029909:Sophora / Q000201:enzymology / Q000235:genetics; D013025:Glycine max / Q000201:enzymology* / Q000235:genetics; D013302:Streptomyces / Q000201:enzymology / Q000235:genetics; D013379:Substrate Specificity
chemical_list: D044950:Flavanones; D059808:Polyphenols; D004122:Dimethylallyltranstransferase; C005273:naringenin