title: Biotransformation of hyoscyamine into scopolamine in transgenic tobacco cell cultures.
abstract: Hyoscyamine-6beta-hydroxylase (H6H) catalyses the conversion of hyoscyamine into its epoxide scopolamine, a compound with a higher added value in the pharmaceutical market than hyoscyamine. We report the establishment of tobacco cell cultures carrying the Hyoscyamus muticus h6h gene under the control of the promoter CAMV 35S. The cell cultures were derived from hairy roots obtained via genetically modified Agrobacterium rhizogenes carrying the pRi and pLAL21 plasmids. The cultures were fed with hyoscyamine, and 4 weeks later the amount of scopolamine produced was quantified by HPLC. The transgenic cell suspension cultures showed a considerable capacity for the bioconversion of hyoscyamine into scopolamine, and released it to the culture medium. Although the scale-up from shake-flask to bioreactor culture usually results in reduced productivities, our transgenic cells grown in a 5-L turbine stirred tank reactor in a batch mode significantly increased the scopolamine accumulation.
authors.affiliation: Departament de Ciències Experimentals i de la Salut, Universitat Pompeu Fabra, Avda Dr Aiguader 80, E-08003 Barcelona, Spain.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Moyano
authors.forename: Javier
authors.initials: J
authors.lastname: Palazón
authors.forename: Mercedes
authors.initials: M
authors.lastname: Bonfill
authors.forename: Lidia
authors.initials: L
authors.lastname: Osuna
authors.forename: Rosa M
authors.initials: RM
authors.lastname: Cusidó
authors.forename: Kirsi-Marja
authors.initials: KM
authors.lastname: Oksman-Caldentey
authors.forename: M Teresa
authors.initials: MT
authors.lastname: Piñol
mesh_terms: D001285:Atropine / Q000378:metabolism*; D001711:Biotransformation; D002478:Cells, Cultured; D017343:Genes, Plant; D032304:Hyoscyamus / Q000201:enzymology / Q000235:genetics; D006899:Mixed Function Oxygenases / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D012231:Rhizobium / Q000235:genetics; D012601:Scopolamine / Q000096:biosynthesis*; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism*; D014170:Transformation, Genetic
chemical_list: D001285:Atropine; D012601:Scopolamine; D006899:Mixed Function Oxygenases; C052273:hyoscyamine (6S)-dioxygenase