title: Evaluation of tobacco (Nicotiana tabacum L. cv. Petit Havana SR1) hairy roots for the production of geraniol, the first committed step in terpenoid indole alkaloid pathway.
abstract: The terpenoid indole alkaloids are one of the major classes of plant-derived natural products and are well known for their many applications in the pharmaceutical, fragrance and cosmetics industries. Hairy root cultures are useful for the production of plant secondary metabolites because of their genetic and biochemical stability and their rapid growth in hormone-free media. Tobacco (Nicotiana tabacum L. cv. Petit Havana SR1) hairy roots, which do not produce geraniol naturally, were engineered to express a plastid-targeted geraniol synthase gene originally isolated from Valeriana officinalis L. (VoGES). A SPME-GC-MS screening tool was developed for the rapid evaluation of production clones. The GC-MS analysis revealed that the free geraniol content in 20 hairy root clones expressing VoGES was an average of 13.7 μg/g dry weight (DW) and a maximum of 31.3 μg/g DW. More detailed metabolic analysis revealed that geraniol derivatives were present in six major glycoside forms, namely the hexose and/or pentose conjugates of geraniol and hydroxygeraniol, resulting in total geraniol levels of up to 204.3 μg/g DW following deglycosylation. A benchtop-scale process was developed in a 20-L wave-mixed bioreactor eventually yielding hundreds of grams of biomass and milligram quantities of geraniol per cultivation bag.
authors: VTT Technical Research Centre of Finland, P.O. Box 1000, Tietotie 2, 02044-VTT Espoo, Finland. Electronic address: anneli.ritala@vtt.fi.
authors: Anneli
authors: A
authors: Ritala
authors: Laboratory of Plant Physiology, Wageningen UR, P.O. Box 658, 6700 AR Wageningen, The Netherlands.
authors: Lemeng
authors: L
authors: Dong
authors: Zurich University of Applied Sciences, Institute of Biotechnology, Biochemical Engineering and Cell Cultivation Technique, Campus Grüental, Wädenswil, Switzerland.
authors: Nicole
authors: N
authors: Imseng
authors: VTT Technical Research Centre of Finland, P.O. Box 1000, Tietotie 2, 02044-VTT Espoo, Finland.
authors: Tuulikki
authors: T
authors: Seppänen-Laakso
authors: Fraunhofer Institute for Molecular Biology and Applied Ecology (IME), Forckenbeckstrasse 6, 52074 Aachen, Germany.
authors: Nikolay
authors: N
authors: Vasilev
authors: Laboratory of Plant Physiology, Wageningen UR, P.O. Box 658, 6700 AR Wageningen, The Netherlands.
authors: Sander
authors: S
authors: van der Krol
authors: VTT Technical Research Centre of Finland, P.O. Box 1000, Tietotie 2, 02044-VTT Espoo, Finland.
authors: Heiko
authors: H
authors: Rischer
authors: VTT Technical Research Centre of Finland, P.O. Box 1000, Tietotie 2, 02044-VTT Espoo, Finland.
authors: Hannu
authors: H
authors: Maaheimo
authors: VTT Technical Research Centre of Finland, P.O. Box 1000, Tietotie 2, 02044-VTT Espoo, Finland.
authors: Arho
authors: A
authors: Virkki
authors: Zurich University of Applied Sciences, Institute of Biotechnology, Biochemical Engineering and Cell Cultivation Technique, Campus Grüental, Wädenswil, Switzerland.
authors: Johanna
authors: J
authors: Brändli
authors: Fraunhofer Institute for Molecular Biology and Applied Ecology (IME), Forckenbeckstrasse 6, 52074 Aachen, Germany.
authors: Stefan
authors: S
authors: Schillberg
authors: Zurich University of Applied Sciences, Institute of Biotechnology, Biochemical Engineering and Cell Cultivation Technique, Campus Grüental, Wädenswil, Switzerland.
authors: Regine
authors: R
authors: Eibl
authors: Laboratory of Plant Physiology, Wageningen UR, P.O. Box 658, 6700 AR Wageningen, The Netherlands.
authors: Harro
authors: H
authors: Bouwmeester
authors: VTT Technical Research Centre of Finland, P.O. Box 1000, Tietotie 2, 02044-VTT Espoo, Finland.
authors: Kirsi-Marja
authors: KM
authors: Oksman-Caldentey
mesh_terms: D000080462:Acyclic Monoterpenes; D019149:Bioreactors; D018744:DNA, Plant; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics / Q000378:metabolism*; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D018517:Plant Roots / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D046948:Secologanin Tryptamine Alkaloids / Q000378:metabolism; D064210:Secondary Metabolism; D013729:Terpenes / Q000378:metabolism*; D014026:Nicotiana / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D014632:Valerian / Q000235:genetics*
chemical_list: D000080462:Acyclic Monoterpenes; D018744:DNA, Plant; D010940:Plant Proteins; D046948:Secologanin Tryptamine Alkaloids; D013729:Terpenes; D010744:Phosphoric Monoester Hydrolases; C007836:geraniol
keywords: Geraniol; Hairy root; Terpenoid indole alkaloid; Tobacco; Transgenic