Cytochrome P450s from Cynara cardunculus L. CYP71AV9 andCYP71BL5, catalyze distinct hydroxylations in the sesquiterpenelactone biosynthetic pathway

K. Eljounaidi, K. Cankar, C. Comino, A. Moglia, A. Hehn, F. Bourgaud, H.J. Bouwmeester, B. Menin, S. Lanteri, M.J. Beekwilder

Research output: Contribution to journalArticleAcademicpeer-review

24 Citations (Scopus)

Abstract

Cynara cardunculus (Asteraceae) is a cross pollinated perennial crop which includes the two cultivatedtaxa globe artichoke and cultivated cardoon. The leaves of these plants contain high concentrationsof sesquiterpene lactones (STLs) among which cynaropicrin is the most represented, and has recentlyattracted attention because of its therapeutic potential as anti-tumor and anti-photoaging agent. Costuno-lide is considered the common precursor of the STLs and three enzymes are involved in its biosyntheticpathway: i.e. the germacrene A synthase (GAS), the germacrene A oxidase (GAO) and the costunolidesynthase (COS). Here we report on the isolation of two P450 genes, (i.e. CYP71AV9 and CYP71BL5), ina set of ~19,000 C. cardunculus unigenes, and their functional characterization in yeast and in planta.The metabolite analyses revealed that the co-expression of CYP71AV9 together with GAS resulted in thebiosynthesis of germacra-1(10),4,11(13)-trien-12-oic acid in yeast. The co-expression of CYP71BL5 andCYP71AV9 with GAS led to biosynthesis of the free costunolide in yeast and costunolide conjugates inNicotiana benthamiana, demonstrating their involvement in STL biosynthesis as GAO and COS enzymes.The substrate specificity of CYP71AV9 was investigated by testing its ability to convert amorpha-4,11-diene, (+)-germacrene D and cascarilladiene to their oxidized products when co-expressed in yeast withthe corresponding terpene synthases.
LanguageEnglish
Pages59-68
JournalPlant Science
Volume223
DOIs
Publication statusPublished - 2014

Fingerprint

Cynara
Cynara cardunculus
germacrene
Biosynthetic Pathways
hydroxylation
Cytochromes
Hydroxylation
cytochromes
biochemical pathways
Sesquiterpenes
Lactones
Yeasts
sesquiterpenoid lactones
yeasts
Oxidoreductases
Cynara scolymus
photoaging
Amorpha
Asteraceae
biosynthesis

Keywords

  • artichoke leaf extract
  • globe artichoke
  • scolymus l.
  • biological-activities
  • yeast expression
  • cynaropicrin
  • synthase
  • chicory
  • (+)-germacrene
  • costunolide

Cite this

Eljounaidi, K. ; Cankar, K. ; Comino, C. ; Moglia, A. ; Hehn, A. ; Bourgaud, F. ; Bouwmeester, H.J. ; Menin, B. ; Lanteri, S. ; Beekwilder, M.J. / Cytochrome P450s from Cynara cardunculus L. CYP71AV9 andCYP71BL5, catalyze distinct hydroxylations in the sesquiterpenelactone biosynthetic pathway. In: Plant Science. 2014 ; Vol. 223. pp. 59-68.
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abstract = "Cynara cardunculus (Asteraceae) is a cross pollinated perennial crop which includes the two cultivatedtaxa globe artichoke and cultivated cardoon. The leaves of these plants contain high concentrationsof sesquiterpene lactones (STLs) among which cynaropicrin is the most represented, and has recentlyattracted attention because of its therapeutic potential as anti-tumor and anti-photoaging agent. Costuno-lide is considered the common precursor of the STLs and three enzymes are involved in its biosyntheticpathway: i.e. the germacrene A synthase (GAS), the germacrene A oxidase (GAO) and the costunolidesynthase (COS). Here we report on the isolation of two P450 genes, (i.e. CYP71AV9 and CYP71BL5), ina set of ~19,000 C. cardunculus unigenes, and their functional characterization in yeast and in planta.The metabolite analyses revealed that the co-expression of CYP71AV9 together with GAS resulted in thebiosynthesis of germacra-1(10),4,11(13)-trien-12-oic acid in yeast. The co-expression of CYP71BL5 andCYP71AV9 with GAS led to biosynthesis of the free costunolide in yeast and costunolide conjugates inNicotiana benthamiana, demonstrating their involvement in STL biosynthesis as GAO and COS enzymes.The substrate specificity of CYP71AV9 was investigated by testing its ability to convert amorpha-4,11-diene, (+)-germacrene D and cascarilladiene to their oxidized products when co-expressed in yeast withthe corresponding terpene synthases.",
keywords = "artichoke leaf extract, globe artichoke, scolymus l., biological-activities, yeast expression, cynaropicrin, synthase, chicory, (+)-germacrene, costunolide",
author = "K. Eljounaidi and K. Cankar and C. Comino and A. Moglia and A. Hehn and F. Bourgaud and H.J. Bouwmeester and B. Menin and S. Lanteri and M.J. Beekwilder",
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Cytochrome P450s from Cynara cardunculus L. CYP71AV9 andCYP71BL5, catalyze distinct hydroxylations in the sesquiterpenelactone biosynthetic pathway. / Eljounaidi, K.; Cankar, K.; Comino, C.; Moglia, A.; Hehn, A.; Bourgaud, F.; Bouwmeester, H.J.; Menin, B.; Lanteri, S.; Beekwilder, M.J.

In: Plant Science, Vol. 223, 2014, p. 59-68.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Cytochrome P450s from Cynara cardunculus L. CYP71AV9 andCYP71BL5, catalyze distinct hydroxylations in the sesquiterpenelactone biosynthetic pathway

AU - Eljounaidi, K.

AU - Cankar, K.

AU - Comino, C.

AU - Moglia, A.

AU - Hehn, A.

AU - Bourgaud, F.

AU - Bouwmeester, H.J.

AU - Menin, B.

AU - Lanteri, S.

AU - Beekwilder, M.J.

PY - 2014

Y1 - 2014

N2 - Cynara cardunculus (Asteraceae) is a cross pollinated perennial crop which includes the two cultivatedtaxa globe artichoke and cultivated cardoon. The leaves of these plants contain high concentrationsof sesquiterpene lactones (STLs) among which cynaropicrin is the most represented, and has recentlyattracted attention because of its therapeutic potential as anti-tumor and anti-photoaging agent. Costuno-lide is considered the common precursor of the STLs and three enzymes are involved in its biosyntheticpathway: i.e. the germacrene A synthase (GAS), the germacrene A oxidase (GAO) and the costunolidesynthase (COS). Here we report on the isolation of two P450 genes, (i.e. CYP71AV9 and CYP71BL5), ina set of ~19,000 C. cardunculus unigenes, and their functional characterization in yeast and in planta.The metabolite analyses revealed that the co-expression of CYP71AV9 together with GAS resulted in thebiosynthesis of germacra-1(10),4,11(13)-trien-12-oic acid in yeast. The co-expression of CYP71BL5 andCYP71AV9 with GAS led to biosynthesis of the free costunolide in yeast and costunolide conjugates inNicotiana benthamiana, demonstrating their involvement in STL biosynthesis as GAO and COS enzymes.The substrate specificity of CYP71AV9 was investigated by testing its ability to convert amorpha-4,11-diene, (+)-germacrene D and cascarilladiene to their oxidized products when co-expressed in yeast withthe corresponding terpene synthases.

AB - Cynara cardunculus (Asteraceae) is a cross pollinated perennial crop which includes the two cultivatedtaxa globe artichoke and cultivated cardoon. The leaves of these plants contain high concentrationsof sesquiterpene lactones (STLs) among which cynaropicrin is the most represented, and has recentlyattracted attention because of its therapeutic potential as anti-tumor and anti-photoaging agent. Costuno-lide is considered the common precursor of the STLs and three enzymes are involved in its biosyntheticpathway: i.e. the germacrene A synthase (GAS), the germacrene A oxidase (GAO) and the costunolidesynthase (COS). Here we report on the isolation of two P450 genes, (i.e. CYP71AV9 and CYP71BL5), ina set of ~19,000 C. cardunculus unigenes, and their functional characterization in yeast and in planta.The metabolite analyses revealed that the co-expression of CYP71AV9 together with GAS resulted in thebiosynthesis of germacra-1(10),4,11(13)-trien-12-oic acid in yeast. The co-expression of CYP71BL5 andCYP71AV9 with GAS led to biosynthesis of the free costunolide in yeast and costunolide conjugates inNicotiana benthamiana, demonstrating their involvement in STL biosynthesis as GAO and COS enzymes.The substrate specificity of CYP71AV9 was investigated by testing its ability to convert amorpha-4,11-diene, (+)-germacrene D and cascarilladiene to their oxidized products when co-expressed in yeast withthe corresponding terpene synthases.

KW - artichoke leaf extract

KW - globe artichoke

KW - scolymus l.

KW - biological-activities

KW - yeast expression

KW - cynaropicrin

KW - synthase

KW - chicory

KW - (+)-germacrene

KW - costunolide

U2 - 10.1016/j.plantsci.2014.03.007

DO - 10.1016/j.plantsci.2014.03.007

M3 - Article

VL - 223

SP - 59

EP - 68

JO - Plant Science

T2 - Plant Science

JF - Plant Science

SN - 0168-9452

ER -