Biochemical and molecular characterization of [alpha]-D-galactosidase from coffee beans

Marraccini Pierre, Rogers William John, Caillet Victoria, Deshayes Alain, Granato Dominique, Lausanne Françoise, Lechat Sylvianne, Pridmore David, Pétiard Vincent. 2005. Biochemical and molecular characterization of [alpha]-D-galactosidase from coffee beans. Plant Physiology and Biochemistry, 43 : pp. 909-920.

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Abstract : [alpha]-D-Galactosidase ([alpha]-Gal; EC is one of three principal enzymes involved in the modification or degradation of plant cell wall galactomannans. In the present paper it is shown that [alpha]-galactosidase activities in field-grown coffee beans are variable amongst cultivars of the two species investigated (Coffea arabica and C. canephora var. Robusta). Higher activities were found in Arabica cultivars. Using beans from greenhouse-cultivated C. arabica as a model, we showed that [alpha]-Gal activity was undetectable in the bean perispem tissue, but increased gradually during the endosperm development, to reach a peak at approximately 30 weeks after flowering (WAF) which coincided with the hardening of the endosperm. [alpha]-Gal-specific transcripts detected at 22 and 27 WAF accompanied the peak of [alpha]-Gal activity, but were reduced to be undetectable in mature beans at 30 WAF, while [alpha]-Gal activity still persisted. Two isoforms were distinguished in 2-DE profiles of crude protein extracts by N-terminal sequencing analysis. Analysis of two-dimensional gel electrophoresis profiles demonstrated that both isoforms accumulated in a linear fashion throughout grain maturation. [alpha]-Gal activity was also observed to increase to high levels during in vitro germination of coffee beans suggesting an important function of this enzyme in this process. [alpha]-Gal cDNA sequences from Arabica and Robusta were sequenced and their deduced proteins appeared to be very similar, differing by only eight amino acids. Southern-blot analysis suggests that the enzyme was encoded by at least two genes in C. arabica that could explain the existence of the two isoforms identified in 2-DE profiles. (Résumé d'auteur)

Mots-clés Agrovoc : Coffea arabica, Coffea canephora, Alpha galactosidase, Mannane, Paroi cellulaire, Fève de café, Développement biologique, Germination, Activité enzymatique, Variété

Mots-clés complémentaires : Cultivar

Classification Agris : F60 - Plant physiology and biochemistry
F30 - Plant genetics and breeding

Champ stratégique Cirad : Axe 1 (2005-2013) - Intensification écologique

Auteurs et affiliations

  • Marraccini Pierre, CIRAD-AMIS-UMR PIA (BRA) ORCID: 0000-0001-7637-6811
  • Rogers William John, Centre de recherches Nestlé (FRA)
  • Caillet Victoria, Centre de recherches Nestlé (FRA)
  • Deshayes Alain, Centre de recherches Nestlé (FRA)
  • Granato Dominique, NESTEC (CHE)
  • Lausanne Françoise, Centre de recherches Nestlé (FRA)
  • Lechat Sylvianne, Centre de recherches Nestlé (FRA)
  • Pridmore David, NESTEC (CHE)
  • Pétiard Vincent, Centre de recherches Nestlé (FRA)

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