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Functional characterization of two p-coumaroyl ester 3′-hydroxylase genes from coffee tree: evidence of a candidate for chlorogenic acid biosynthesis

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Abstract

Chlorogenic acid (5-CQA) is one of the major soluble phenolic compounds that is accumulated in coffee green beans. With other hydroxycinnamoyl quinic acids (HQAs), this compound is accumulated in particular in green beans of the cultivated species Coffea canephora. Recent work has indicated that the biosynthesis of 5-CQA can be catalyzed by a cytochrome P450 enzyme, CYP98A3 from Arabidopsis. Two full-length cDNA clones (CYP98A35 and CYP98A36) that encode putative p-coumaroylester 3′-hydroxylases (C3′H) were isolated from C. canephora cDNA libraries. Recombinant protein expression in yeast showed that both metabolized p-coumaroyl shikimate at similar rates, but that only one hydroxylates the chlorogenic acid precursor p-coumaroyl quinate. CYP98A35 appears to be the first C3′H capable of metabolising p-coumaroyl quinate and p-coumaroyl shikimate with the same efficiency. We studied the expression patterns of both genes on 4-month old C. canephora plants and found higher transcript levels in young and in highly vascularized organs for both genes. Gene expression and HQA content seemed to be correlated in these organs. Histolocalization and immunolocalization studies revealed similar tissue localization for caffeoyl quinic acids and p-coumaroylester 3′-hydroxylases. The results indicated that HQA biosynthesis and accumulation occurred mainly in the shoot tip and in the phloem of the vascular bundles. The lack of correlation between gene expression and HQA content observed in some organs is discussed in terms of transport and accumulation mechanisms.

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Abbreviations

CQA:

caffeoyl quinic acid

5-CQA:

chlorogenic acid

HQA:

hydroxycinnamoyl quinic acid

C3′H:

p-coumaroyl ester 3′-hydroxylases

HQT:

hydroxycinnamoyl-CoA: quinate hydroxycinnamoyl transferase

HCT:

hydroxycinnamoyl-CoA: shikimate/quinate hydroxycinnamoyl transferase

CODEHOP:

consensus degenerate hybrid oligonucleotide primers

CYP:

cytochrome P450 monooxygenases

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Acknowledgements

Mahesh V. was granted by Avesthagen graine and J. Bustamante by INIA from Venezuela

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Authors and Affiliations

  1. Laboratoire de Génomique et Qualité du café, IRD, UMR 1097 DGPC, 911 Avenue Agropolis, BP 64501, 34394, Montpellier cedex 5, France

    Venkataramaiah Mahesh, Nathalie Chabrillange, José Bustamante, Michel Noirot, Serge Hamon, Alexandre de Kochko & Claudine Campa

  2. Department of Plant Stress Response, Institute of Plant Molecular Biology, CNRS-UPR 2357, Université Louis Pasteur, 28 rue Goethe, 67083, Strasbourg, France

    Rachel Million-Rousseau, Pascaline Ullmann & Danièle Werck-Reichhart

  3. Laboratoire de Botanique, Phytochimie et Mycologie, UMR 5175 CEFE-CNRS, Faculté de Pharmacie, 15 av. Charles Flahault, BP 14491, 34093, Montpellier cedex 5, France

    Laurence Mondolot

  4. Avesthagen graine, A Plant Genome Biology Laboratory, 9th floor, Discoverer, ITPL, Bangalore, India

    Venkataramaiah Mahesh

  5. Department of Plant Biology, Thorvaldsensvej 40, 1871, Fredericksberg C, Copenhagen, Denmark

    Marc Morant

Authors
  1. Venkataramaiah Mahesh
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  2. Rachel Million-Rousseau
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  3. Pascaline Ullmann
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  4. Nathalie Chabrillange
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  5. José Bustamante
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  6. Laurence Mondolot
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  7. Marc Morant
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  8. Michel Noirot
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  9. Serge Hamon
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  10. Alexandre de Kochko
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  11. Danièle Werck-Reichhart
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  12. Claudine Campa
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Corresponding author

Correspondence to Claudine Campa.

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Mahesh, V., Million-Rousseau, R., Ullmann, P. et al. Functional characterization of two p-coumaroyl ester 3′-hydroxylase genes from coffee tree: evidence of a candidate for chlorogenic acid biosynthesis. Plant Mol Biol 64, 145–159 (2007). https://doi.org/10.1007/s11103-007-9141-3

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  • DOI: https://doi.org/10.1007/s11103-007-9141-3

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