Plant Molecular Biology

, Volume 64, Issue 1, pp 145–159

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

  • Venkataramaiah Mahesh
  • Rachel Million-Rousseau
  • Pascaline Ullmann
  • Nathalie Chabrillange
  • José Bustamante
  • Laurence Mondolot
  • Marc Morant
  • Michel Noirot
  • Serge Hamon
  • Alexandre de Kochko
  • Danièle Werck-Reichhart
  • Claudine Campa
Article

DOI: 10.1007/s11103-007-9141-3

Cite this article as:
Mahesh, V., Million-Rousseau, R., Ullmann, P. et al. Plant Mol Biol (2007) 64: 145. doi:10.1007/s11103-007-9141-3

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.

Keywords

Caffeoyl quinic acidsChlorogenic acid Coffea canephoraCytochrome P450 hydroxylase

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

Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • Venkataramaiah Mahesh
    • 1
    • 4
  • Rachel Million-Rousseau
    • 2
  • Pascaline Ullmann
    • 2
  • Nathalie Chabrillange
    • 1
  • José Bustamante
    • 1
  • Laurence Mondolot
    • 3
  • Marc Morant
    • 5
  • Michel Noirot
    • 1
  • Serge Hamon
    • 1
  • Alexandre de Kochko
    • 1
  • Danièle Werck-Reichhart
    • 2
  • Claudine Campa
    • 1
  1. 1.Laboratoire de Génomique et Qualité du caféIRD, UMR 1097 DGPCMontpellier cedex 5France
  2. 2.Department of Plant Stress Response, Institute of Plant Molecular Biology, CNRS-UPR 2357Université Louis PasteurStrasbourgFrance
  3. 3.Laboratoire de Botanique, Phytochimie et MycologieUMR 5175 CEFE-CNRS, Faculté de PharmacieMontpellier cedex 5France
  4. 4.Avesthagen graineA Plant Genome Biology LaboratoryBangaloreIndia
  5. 5.Department of Plant BiologyFredericksberg CDenmark