Plant Molecular Biology

, 72:17 | Cite as

A novel 4-hydroxycoumarin biosynthetic pathway

  • Benye Liu
  • Torben Raeth
  • Till Beuerle
  • Ludger Beerhues


Coumarin forms in melilotoside (trans-ortho-coumaric acid glucoside)-containing plant species upon cell damage. In moldy melilotoside-containing plant material, trans-ortho-coumaric acid is converted by fungi to 4-hydroxycoumarin, two molecules of which spontaneously combine with formaldehyde to give dicoumarol. Dicoumarol causes internal bleeding in livestock and is the forerunner of the warfarin group of medicinal anticoagulants. Here, we report 4-hydroxycoumarin formation by biphenyl synthase (BIS). Two new BIS cDNAs were isolated from elicitor-treated Sorbus aucuparia cell cultures. The encoded isoenzymes preferred ortho-hydroxybenzoyl (salicoyl)-CoA as a starter substrate and catalyzed a single decarboxylative condensation with malonyl-CoA to give 4-hydroxycoumarin. When elicitor-treated S. aucuparia cell cultures were fed with the N-acetylcysteamine thioester of salicylic acid, 4-hydroxycoumarin accumulated in the culture medium. Incubation of the BIS isoenzymes with benzoyl-CoA and malonyl-CoA resulted in the formation of 3,5-dihydroxybiphenyl which is the precursor of the phytoalexins of the Maloideae.


4-Hydroxycoumarin Biphenyl synthase Type III polyketide synthase Salicylic acid Salicoyl-NAC Sorbus aucuparia 



This work was supported by a grant from the Deutsche Forschungsgemeinschaft (focus program 1152).


  1. Abd El-Mawla AMA, Beerhues L (2002) Benzoic acid biosynthesis in cell cultures of Hypericum androsaemum. Planta 214:727–733CrossRefPubMedGoogle Scholar
  2. Abe I, Abe T, Wanibuchi K, Noguchi H (2006) Enzymatic formation of quinolone alkaloids by a plant type III polyketide synthase. Org Lett 8:6063–6065CrossRefPubMedGoogle Scholar
  3. Aliotta G, Cafiero G, De Feo V, Sacchi R (1994) Potential allelochemicals from Ruta graveolens L. and their action on radish seeds. J Chem Ecol 20:2761–2775CrossRefGoogle Scholar
  4. Appendino G, Tagliapietra S, Nano GM, Picci V (1988a) Ferprenin, a prenylated coumarin from Ferula communis. Phytochemistry 27:944–946CrossRefGoogle Scholar
  5. Appendino G, Tagliapietra S, Gariboldi P, Nano GM, Picci V (1988b) ω-Oxygenated prenylated coumarins from Ferula communis. Phytochemistry 27:3619–3624CrossRefGoogle Scholar
  6. Arnoldi L, Ballero M, Fuzzati N, Maxia A, Percalli E, Pagni L (2004) HPLC-DAD-MS identification of bioactive secondary metabolites from Ferula communis roots. Fitoterapia 75:342–354CrossRefPubMedGoogle Scholar
  7. Austin MB, Noel JP (2003) The chalcone synthase superfamily of type III polyketide synthases. Nat Prod Rep 20:79–110CrossRefPubMedGoogle Scholar
  8. Beuerle T, Pichersky E (2002) Purification and characterization of benzoate: coenzyme A ligase from Clarkia breweri. Arch Biochem Biophys 400:258–264CrossRefPubMedGoogle Scholar
  9. Bourgaud F, Hehn A, Larbat R, Doerper S, Gontier E, Kellner S, Matern U (2006) Biosynthesis of coumarins in plants: a major pathway still to be unravelled for cytochrome P450 enzymes. Phytochem Rev 5:293–308CrossRefGoogle Scholar
  10. Brown SA (1986) Biochemistry of plant coumarins. In: Conn EE (ed) Recent advances in phytochemistry, vol 20: the shikimic acid pathway. Plenum Press, New York, pp 287–316Google Scholar
  11. Bye A, King HK (1970) The biosynthesis of 4-hydroxycoumarin and dicoumarol by Aspergillus fumigatus Fresenius. Biochem J 117:237–245PubMedGoogle Scholar
  12. Chexal KK, Fouweather C, Holker JS (1975) The biosynthesis of fungal metabolites. Part VII. Production and biosynthesis of 4,7-dimethoxy-5-methylcoumarin in Aspergillus variecolor. J Chem Soc (Perkin 1):554–556Google Scholar
  13. Ferrer JL, Jez JM, Bowman ME, Dixon RA, Noel JP (1999) Structure of chalcone synthase and the molecular basis of plant polyketide biosynthesis. Nat Struct Biol 6:775–784CrossRefPubMedGoogle Scholar
  14. Gaid MM, Sircar D, Beuerle T, Mitra A, Beerhues L (2009) Benzaldehyde dehydrogenase from chitosan-treated Sorbus aucuparia cell cultures. J Plant Physiol 166:1343–1349CrossRefPubMedGoogle Scholar
  15. Inoue T, Toyonaga T, Nagumo S, Nagai M (1989) Biosynthesis of 4-hydroxy-5-methylcoumarin in a Gerbera jamesonii hybrid. Phytochemistry 28:2329–2330CrossRefGoogle Scholar
  16. Jez JM, Bowman ME, Noel JP (2002) Expanding the biosynthetic repertoire of plant type III polyketide synthases by altering starter molecule specificity. Proc Natl Acad Sci USA 99:5319–5324CrossRefPubMedGoogle Scholar
  17. Kurosaki F, Itoh M, Kizawa Y, Nishi A (1993) Partial purification and characterization of a polyketide biosynthetic enzyme, 6-hydroxymellein synthase, in elicitor-treated carrot cell extracts. Arch Biochem Biophys 300:157–163CrossRefPubMedGoogle Scholar
  18. Liu B, Falkenstein-Paul H, Schmidt W, Beerhues L (2003) Benzophenone synthase and chalcone synthase from Hypericum androsaemum cell cultures: cDNA cloning, functional expression, and site-directed mutagenesis of two polyketide synthases. Plant J 34:847–855CrossRefPubMedGoogle Scholar
  19. Liu B, Beuerle T, Klundt T, Beerhues L (2004) Biphenyl synthase from yeast-extract-treated cell cultures of Sorbus aucuparia. Planta 218:492–496CrossRefPubMedGoogle Scholar
  20. Liu B, Raeth T, Beuerle T, Beerhues L (2007) Biphenyl synthase, a novel type III polyketide synthase. Planta 225:1495–1503CrossRefPubMedGoogle Scholar
  21. Matern U (1991) Coumarins and other phenylpropanoid compounds in the defense response of plant cells. Planta Med 57(Suppl 1):S15–S20CrossRefPubMedGoogle Scholar
  22. Matern U, Lüer P, Kreusch D (1999) Biosynthesis of coumarins. In: Sankawa U (ed) Comprehensive natural products chemistry, vol 1: polyketides and other secondary metabolites including fatty acids and their derivatives. Elsevier, Amsterdam, pp 623–637Google Scholar
  23. Murray RDH (1995) Coumarins. Nat Prod Rep 12:477–505CrossRefGoogle Scholar
  24. Murray RDH, Méndez J, Brown SA (1982) The natural coumarins: occurrence, chemistry and biochemistry. Wiley, ChichesterGoogle Scholar
  25. Oguro S, Akashi T, Ayabe S, Noguchi H, Abe I (2004) Probing biosynthesis of plant polyketides with synthetic N-acetylcysteamine thioesters. Biochem Biophys Res Commun 325:561–567CrossRefPubMedGoogle Scholar
  26. Späth E, Simon AFJ, Lintner J (1936) Die Konstitution des Ammoresinols (XIX. Mitteil. über natürliche Cumarine). Ber Dt Chem Ges 69:1656–1664CrossRefGoogle Scholar
  27. Suo Z, Chen H, Walsh CT (2000) Acyl-CoA hydrolysis by the high molecular weight protein 1 subunit of yersiniabactin synthetase: mutational evidence for a cascade of four acyl-enzyme intermediates during hydrolytic editing. Proc Natl Acad Sci USA 97:14188–14193CrossRefPubMedGoogle Scholar
  28. Valle MG, Appendino G, Nano GM, Picci V (1987) Prenylated coumarins and sesquiterpenoids from Ferula communis. Phytochemistry 26:253–256CrossRefGoogle Scholar
  29. Wildermuth MC (2006) Variations on a theme: synthesis and modification of plant benzoic acids. Curr Opin Plant Biol 9:288–296CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Benye Liu
    • 1
    • 2
  • Torben Raeth
    • 1
  • Till Beuerle
    • 1
  • Ludger Beerhues
    • 1
  1. 1.Institut für Pharmazeutische BiologieBraunschweigGermany
  2. 2.Key Laboratory of Photosynthesis and Environmental Molecular PhysiologyInstitute of Botany, The Chinese Academy of SciencesBeijingChina

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