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

详细信息    查看全文

• 作者：Anneli Ritala^a ; ^{anneli.ritala@vtt.fi" class="auth_mail} ; Lemeng Dong^b ; Nicole Imseng^c ; Tuulikki Seppä ; nen-Laakso^a ; Nikolay Vasilev^d ; Sander van der Krol^b ; Heiko Rischer^a ; Hannu Maaheimo^a ; Arho Virkki^a ; Johanna Brä ; ndli^c ; Stefan Schillberg^d ; Regine Eibl^c ; Harro Bouwmeester^b ; Kirsi-Marja Oksman-Caldentey^a
• 关键词：Geraniol ; Hairy root ; Terpenoid indole alkaloid ; Tobacco ; Transgenic
• 刊名：Journal of Biotechnology
• 出版年：20 April, 2014
• 年：2014
• 卷：176
• 期：Complete
• 页码：20-28
• 全文大小：1463 K

文摘

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 渭g/g dry weight (DW) and a maximum of 31.3 渭g/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 渭g/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.