Biosynthesis of podophyllotoxin in Linum album cell cultures

Veronique Seidel, Jorg Windhövel, Graham Eaton, A. Wilhelm Alfermann, Randolph R.J. Arroo, Manuel Medarde, Maike Petersen, Jack G. Woolley

Research output: Contribution to journalArticle

56 Citations (Scopus)

Abstract

Cell cultures of Linum album Kotschy ex Boiss. (Linaceae) showing high accumulation of the lignan podophyllotoxin (PTOX) were established. Enzymological studies revealed highest activities of phenylalanine ammonia-lyase, cinnamyl alcohol dehydrogenase, 4-hydroxycinnamate:CoA ligase and cinnamoyl-CoA:-NADP oxidoreductase immediately prior to PTOX accumulation. To investigate PTOX biosynthesis, feeding experiments were performed with [2- 13 C]3¢,4¢-dimethoxycinnamic acid, [2-13 C]3¢,4¢-methylenedioxycinnamic acid (MDCA), [2-13C]3¢,4¢,5¢-trimethoxycinnamic acid,[2- 13C]sinapic acid, [2-13C]- and [2,3-13C2]ferulic acid.Analysis of the metabolites by HPLC coupled to tandem mass spectrometry revealed incorporation of label from ferulic acid into PTOX and deoxypodophyllotoxin (DOP). In addition, MDCA was also unambiguously incorporated intact into PTOX. These observations suggest that in L. album both ferulic acid and methylenedioxy-substituted cinnamic acid can be incorporated into lignans. Furthermore, it appears that, in this species, the hydroxylation of DOP is a rate-limiting point in the pathway leading to PTOX.
LanguageEnglish
Pages1031-1039
Number of pages9
JournalPlanta
Volume215
Issue number6
DOIs
Publication statusPublished - 1 Oct 2002

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Podophyllotoxin
Flax
ferulic acid
cell culture
Cell Culture Techniques
biosynthesis
sinapic acid
Lignans
acids
lignans
Linaceae
Phenylalanine Ammonia-Lyase
cinnamyl alcohol dehydrogenase
Acids
cinnamic acid
oxidoreductases
hydroxylation
phenylalanine ammonia-lyase
Hydroxylation
Tandem Mass Spectrometry

Keywords

  • podophyllotoxin
  • biosynthesis
  • linum album

Cite this

Seidel, V., Windhövel, J., Eaton, G., Alfermann, A. W., Arroo, R. R. J., Medarde, M., ... Woolley, J. G. (2002). Biosynthesis of podophyllotoxin in Linum album cell cultures. Planta, 215(6), 1031-1039. https://doi.org/10.1007/s00425-002-0834-1
Seidel, Veronique ; Windhövel, Jorg ; Eaton, Graham ; Alfermann, A. Wilhelm ; Arroo, Randolph R.J. ; Medarde, Manuel ; Petersen, Maike ; Woolley, Jack G. / Biosynthesis of podophyllotoxin in Linum album cell cultures. In: Planta. 2002 ; Vol. 215, No. 6. pp. 1031-1039.
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Seidel, V, Windhövel, J, Eaton, G, Alfermann, AW, Arroo, RRJ, Medarde, M, Petersen, M & Woolley, JG 2002, 'Biosynthesis of podophyllotoxin in Linum album cell cultures' Planta, vol. 215, no. 6, pp. 1031-1039. https://doi.org/10.1007/s00425-002-0834-1

Biosynthesis of podophyllotoxin in Linum album cell cultures. / Seidel, Veronique; Windhövel, Jorg; Eaton, Graham; Alfermann, A. Wilhelm; Arroo, Randolph R.J.; Medarde, Manuel; Petersen, Maike; Woolley, Jack G.

In: Planta, Vol. 215, No. 6, 01.10.2002, p. 1031-1039.

Research output: Contribution to journalArticle

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T1 - Biosynthesis of podophyllotoxin in Linum album cell cultures

AU - Seidel, Veronique

AU - Windhövel, Jorg

AU - Eaton, Graham

AU - Alfermann, A. Wilhelm

AU - Arroo, Randolph R.J.

AU - Medarde, Manuel

AU - Petersen, Maike

AU - Woolley, Jack G.

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N2 - Cell cultures of Linum album Kotschy ex Boiss. (Linaceae) showing high accumulation of the lignan podophyllotoxin (PTOX) were established. Enzymological studies revealed highest activities of phenylalanine ammonia-lyase, cinnamyl alcohol dehydrogenase, 4-hydroxycinnamate:CoA ligase and cinnamoyl-CoA:-NADP oxidoreductase immediately prior to PTOX accumulation. To investigate PTOX biosynthesis, feeding experiments were performed with [2- 13 C]3¢,4¢-dimethoxycinnamic acid, [2-13 C]3¢,4¢-methylenedioxycinnamic acid (MDCA), [2-13C]3¢,4¢,5¢-trimethoxycinnamic acid,[2- 13C]sinapic acid, [2-13C]- and [2,3-13C2]ferulic acid.Analysis of the metabolites by HPLC coupled to tandem mass spectrometry revealed incorporation of label from ferulic acid into PTOX and deoxypodophyllotoxin (DOP). In addition, MDCA was also unambiguously incorporated intact into PTOX. These observations suggest that in L. album both ferulic acid and methylenedioxy-substituted cinnamic acid can be incorporated into lignans. Furthermore, it appears that, in this species, the hydroxylation of DOP is a rate-limiting point in the pathway leading to PTOX.

AB - Cell cultures of Linum album Kotschy ex Boiss. (Linaceae) showing high accumulation of the lignan podophyllotoxin (PTOX) were established. Enzymological studies revealed highest activities of phenylalanine ammonia-lyase, cinnamyl alcohol dehydrogenase, 4-hydroxycinnamate:CoA ligase and cinnamoyl-CoA:-NADP oxidoreductase immediately prior to PTOX accumulation. To investigate PTOX biosynthesis, feeding experiments were performed with [2- 13 C]3¢,4¢-dimethoxycinnamic acid, [2-13 C]3¢,4¢-methylenedioxycinnamic acid (MDCA), [2-13C]3¢,4¢,5¢-trimethoxycinnamic acid,[2- 13C]sinapic acid, [2-13C]- and [2,3-13C2]ferulic acid.Analysis of the metabolites by HPLC coupled to tandem mass spectrometry revealed incorporation of label from ferulic acid into PTOX and deoxypodophyllotoxin (DOP). In addition, MDCA was also unambiguously incorporated intact into PTOX. These observations suggest that in L. album both ferulic acid and methylenedioxy-substituted cinnamic acid can be incorporated into lignans. Furthermore, it appears that, in this species, the hydroxylation of DOP is a rate-limiting point in the pathway leading to PTOX.

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Seidel V, Windhövel J, Eaton G, Alfermann AW, Arroo RRJ, Medarde M et al. Biosynthesis of podophyllotoxin in Linum album cell cultures. Planta. 2002 Oct 1;215(6):1031-1039. https://doi.org/10.1007/s00425-002-0834-1