(+)-Valencene production in Nicotiana benthamiana is increased by down-regulation of competing pathways

K. Cankar, E.J. Jongedijk, M. Klompmaker, T. Majdic, R. Mumm, H.J. Bouwmeester, H.J. Bosch, M.J. Beekwilder

Research output: Contribution to journalArticleAcademicpeer-review

17 Citations (Scopus)

Abstract

Plant sesquiterpenes, such as (+)-valencene, artemisinin, and farnesene are valuable chemicals for use as aromatics, pharmaceuticals, and biofuels. Plant-based production systems for terpenoids critically depend on the availability of farnesyl diphosphate (FPP). Currently, these systems show insufficient yields, due to the competition for FPP of newly introduced pathways with endogenous ones. In this study, for the first time an RNAi strategy aiming at silencing of endogenous pathways for increased (+)-valencene production was employed. Firstly, a transient production system for (+)-valencene in Nicotiana benthamiana was set up using agroinfiltration. Secondly, silencing of the endogenous 5-epi-aristolochene synthase (EAS) and squalene synthase (SQS) that compete for the FPP pool was deployed. This resulted in a N. benthamiana plant that produces (+)-valencene as a prevalent volatile with a 2.8-fold increased yield. Finally, the size of the FPP pool was increased by overexpression of enzymes that are rate-limiting in FPP biosynthesis. Combined with silencing of EAS and SQS, no further increase of (+)-valencene production was observed, but emission of farnesol. Formation of farnesol, which is a breakdown product of FPP, indicates that overproducing sesquiterpenes is no longer limited by FPP availability in the cytosol. This study shows that metabolic engineering of plants can effectively be used for increased production of desired products in plants. Keywords: 5-Epi-aristolochene synthase · Metabolic engineering · RNAi · Squalene synthase
LanguageEnglish
Pages180-189
JournalBiotechnology Journal
Volume10
Issue number1
DOIs
Publication statusPublished - 2015

Fingerprint

Farnesyl-Diphosphate Farnesyltransferase
Tobacco
Down-Regulation
Farnesol
Metabolic Engineering
Sesquiterpenes
RNA Interference
Biofuels
Cytosol
valencene
Enzymes
Pharmaceutical Preparations
5-epi-aristolochene synthase

Keywords

  • plant transformation
  • biosynthetic-pathway
  • terpenoid metabolism
  • squalene synthase
  • tobacco
  • expression
  • artemisinin
  • arabidopsis
  • reductase
  • precursors

Cite this

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title = "(+)-Valencene production in Nicotiana benthamiana is increased by down-regulation of competing pathways",
abstract = "Plant sesquiterpenes, such as (+)-valencene, artemisinin, and farnesene are valuable chemicals for use as aromatics, pharmaceuticals, and biofuels. Plant-based production systems for terpenoids critically depend on the availability of farnesyl diphosphate (FPP). Currently, these systems show insufficient yields, due to the competition for FPP of newly introduced pathways with endogenous ones. In this study, for the first time an RNAi strategy aiming at silencing of endogenous pathways for increased (+)-valencene production was employed. Firstly, a transient production system for (+)-valencene in Nicotiana benthamiana was set up using agroinfiltration. Secondly, silencing of the endogenous 5-epi-aristolochene synthase (EAS) and squalene synthase (SQS) that compete for the FPP pool was deployed. This resulted in a N. benthamiana plant that produces (+)-valencene as a prevalent volatile with a 2.8-fold increased yield. Finally, the size of the FPP pool was increased by overexpression of enzymes that are rate-limiting in FPP biosynthesis. Combined with silencing of EAS and SQS, no further increase of (+)-valencene production was observed, but emission of farnesol. Formation of farnesol, which is a breakdown product of FPP, indicates that overproducing sesquiterpenes is no longer limited by FPP availability in the cytosol. This study shows that metabolic engineering of plants can effectively be used for increased production of desired products in plants. Keywords: 5-Epi-aristolochene synthase · Metabolic engineering · RNAi · Squalene synthase",
keywords = "plant transformation, biosynthetic-pathway, terpenoid metabolism, squalene synthase, tobacco, expression, artemisinin, arabidopsis, reductase, precursors",
author = "K. Cankar and E.J. Jongedijk and M. Klompmaker and T. Majdic and R. Mumm and H.J. Bouwmeester and H.J. Bosch and M.J. Beekwilder",
year = "2015",
doi = "10.1002/biot.201400288",
language = "English",
volume = "10",
pages = "180--189",
journal = "Biotechnology Journal",
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(+)-Valencene production in Nicotiana benthamiana is increased by down-regulation of competing pathways. / Cankar, K.; Jongedijk, E.J.; Klompmaker, M.; Majdic, T.; Mumm, R.; Bouwmeester, H.J.; Bosch, H.J.; Beekwilder, M.J.

In: Biotechnology Journal, Vol. 10, No. 1, 2015, p. 180-189.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - (+)-Valencene production in Nicotiana benthamiana is increased by down-regulation of competing pathways

AU - Cankar, K.

AU - Jongedijk, E.J.

AU - Klompmaker, M.

AU - Majdic, T.

AU - Mumm, R.

AU - Bouwmeester, H.J.

AU - Bosch, H.J.

AU - Beekwilder, M.J.

PY - 2015

Y1 - 2015

N2 - Plant sesquiterpenes, such as (+)-valencene, artemisinin, and farnesene are valuable chemicals for use as aromatics, pharmaceuticals, and biofuels. Plant-based production systems for terpenoids critically depend on the availability of farnesyl diphosphate (FPP). Currently, these systems show insufficient yields, due to the competition for FPP of newly introduced pathways with endogenous ones. In this study, for the first time an RNAi strategy aiming at silencing of endogenous pathways for increased (+)-valencene production was employed. Firstly, a transient production system for (+)-valencene in Nicotiana benthamiana was set up using agroinfiltration. Secondly, silencing of the endogenous 5-epi-aristolochene synthase (EAS) and squalene synthase (SQS) that compete for the FPP pool was deployed. This resulted in a N. benthamiana plant that produces (+)-valencene as a prevalent volatile with a 2.8-fold increased yield. Finally, the size of the FPP pool was increased by overexpression of enzymes that are rate-limiting in FPP biosynthesis. Combined with silencing of EAS and SQS, no further increase of (+)-valencene production was observed, but emission of farnesol. Formation of farnesol, which is a breakdown product of FPP, indicates that overproducing sesquiterpenes is no longer limited by FPP availability in the cytosol. This study shows that metabolic engineering of plants can effectively be used for increased production of desired products in plants. Keywords: 5-Epi-aristolochene synthase · Metabolic engineering · RNAi · Squalene synthase

AB - Plant sesquiterpenes, such as (+)-valencene, artemisinin, and farnesene are valuable chemicals for use as aromatics, pharmaceuticals, and biofuels. Plant-based production systems for terpenoids critically depend on the availability of farnesyl diphosphate (FPP). Currently, these systems show insufficient yields, due to the competition for FPP of newly introduced pathways with endogenous ones. In this study, for the first time an RNAi strategy aiming at silencing of endogenous pathways for increased (+)-valencene production was employed. Firstly, a transient production system for (+)-valencene in Nicotiana benthamiana was set up using agroinfiltration. Secondly, silencing of the endogenous 5-epi-aristolochene synthase (EAS) and squalene synthase (SQS) that compete for the FPP pool was deployed. This resulted in a N. benthamiana plant that produces (+)-valencene as a prevalent volatile with a 2.8-fold increased yield. Finally, the size of the FPP pool was increased by overexpression of enzymes that are rate-limiting in FPP biosynthesis. Combined with silencing of EAS and SQS, no further increase of (+)-valencene production was observed, but emission of farnesol. Formation of farnesol, which is a breakdown product of FPP, indicates that overproducing sesquiterpenes is no longer limited by FPP availability in the cytosol. This study shows that metabolic engineering of plants can effectively be used for increased production of desired products in plants. Keywords: 5-Epi-aristolochene synthase · Metabolic engineering · RNAi · Squalene synthase

KW - plant transformation

KW - biosynthetic-pathway

KW - terpenoid metabolism

KW - squalene synthase

KW - tobacco

KW - expression

KW - artemisinin

KW - arabidopsis

KW - reductase

KW - precursors

U2 - 10.1002/biot.201400288

DO - 10.1002/biot.201400288

M3 - Article

VL - 10

SP - 180

EP - 189

JO - Biotechnology Journal

T2 - Biotechnology Journal

JF - Biotechnology Journal

SN - 1860-6768

IS - 1

ER -