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

Anneli Ritala, L Dong, N Imseng, Tuulikki Seppänen-Laakso, N Vasilev, S van der Krol, Heiko Rischer, Hannu Maaheimo, Arho Virkki, J Brändli, S Schillerberg, R Eibl, H Bouwmeester, Kirsi-Marja Oksman-Caldentey

Research output: Contribution to journalArticleScientificpeer-review

15 Citations (Scopus)

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.
Original languageEnglish
Pages (from-to)20-28
Number of pages8
JournalJournal of Biotechnology
Volume176
DOIs
Publication statusPublished - 2014
MoE publication typeA1 Journal article-refereed

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Secologanin Tryptamine Alkaloids
Alkaloids
Tobacco
Fragrances
Glycosides
Cosmetics
Hormones
Metabolites
Bioreactors
Drug products
Screening
Biomass
Genes
Derivatives
Weights and Measures
Clone Cells
Industry
Valerian
Pentoses
Plastids

Keywords

  • Geraniol
  • hairy root
  • terpenoid indole alkaloid
  • tobacco
  • transgenic

Cite this

@article{60d587c6fa2d4712b58614525e6c330c,
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.",
keywords = "Geraniol, hairy root, terpenoid indole alkaloid, tobacco, transgenic",
author = "Anneli Ritala and L Dong and N Imseng and Tuulikki Sepp{\"a}nen-Laakso and N Vasilev and {van der Krol}, S and Heiko Rischer and Hannu Maaheimo and Arho Virkki and J Br{\"a}ndli and S Schillerberg and R Eibl and H Bouwmeester and Kirsi-Marja Oksman-Caldentey",
year = "2014",
doi = "10.1016/j.jbiotec.2014.01.031",
language = "English",
volume = "176",
pages = "20--28",
journal = "Journal of Biotechnology",
issn = "0168-1656",
publisher = "Elsevier",

}

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. / Ritala, Anneli; Dong, L; Imseng, N; Seppänen-Laakso, Tuulikki; Vasilev, N; van der Krol, S; Rischer, Heiko; Maaheimo, Hannu; Virkki, Arho; Brändli, J; Schillerberg, S; Eibl, R; Bouwmeester, H; Oksman-Caldentey, Kirsi-Marja.

In: Journal of Biotechnology, Vol. 176, 2014, p. 20-28.

Research output: Contribution to journalArticleScientificpeer-review

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

VL - 176

SP - 20

EP - 28

JO - Journal of Biotechnology

JF - Journal of Biotechnology

SN - 0168-1656

ER -