TY - JOUR
T1 - 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
AU - Ritala, Anneli
AU - Dong, L
AU - Imseng, N
AU - Seppänen-Laakso, Tuulikki
AU - Vasilev, N
AU - van der Krol, S
AU - Rischer, Heiko
AU - Maaheimo, Hannu
AU - Virkki, Arho
AU - Brändli, J
AU - Schillerberg, S
AU - Eibl, R
AU - Bouwmeester, H
AU - Oksman-Caldentey, Kirsi-Marja
PY - 2014
Y1 - 2014
N2 - 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.
AB - 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.
KW - Geraniol
KW - hairy root
KW - terpenoid indole alkaloid
KW - tobacco
KW - transgenic
U2 - 10.1016/j.jbiotec.2014.01.031
DO - 10.1016/j.jbiotec.2014.01.031
M3 - Article
SN - 0168-1656
VL - 176
SP - 20
EP - 28
JO - Journal of Biotechnology
JF - Journal of Biotechnology
ER -