Biotransformation of hyoscyamine into scopolamine in transgenic tobacco cell cultures.
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.
Moyano
Elisabeth
E
Departament de Ciències Experimentals i de la Salut, Universitat Pompeu Fabra, Avda Dr Aiguader 80, E-08003 Barcelona, Spain.
Palazón
Javier
J
Bonfill
Mercedes
M
Osuna
Lidia
L
Cusidó
Rosa M
RM
Oksman-Caldentey
Kirsi-Marja
KM
Piñol
M Teresa
MT
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
D001285:Atropine; D012601:Scopolamine; D006899:Mixed Function Oxygenases; C052273:hyoscyamine (6S)-dioxygenase