Skip to main content Accessibility help

In vivo kinetics of β-glucosidase towards glucovanillin and related phenolic glucosides in heat-treated vanilla pod (Vanilla planifolia, Orchidaceae)

  • Jean-Marc Brillouet (a1) (a2) and Éric Odoux (a1)


Introduction. The traditional curing of vanilla pods includes “killing” and sweating steps when pods are exposed to heat (35–65 ℃) for various lengths of time. Although it is known that liberation of vanillin and other phenolics from their non-aromatic glucosides is due to the action of an endogenous β-glucosidase, its in vivo kinetics remained unknown. Materials and methods. Mature green vanilla pods were pretreated for 2 h at 50 ℃, 55 ℃ and 60 ℃, then stored for 118 days at 27 ℃. Phenolic glucosides and their aglycons were extracted at regular intervals during the storage period and analyzed by HPLC. Results and discussion. All phenolic β-glucosides were slowly hydrolyzed during the storage period with production of vanillin, p-hydroxybenzaldehyde, vanillic acid, and other unknown aglycons. Most of the β-glucosidase was heat-denatured by the pretreatment, and analysis of its kinetic parameters showed that it adopts, in vivo, an allosteric mode of functioning with a lower affinity for glucovanillin than in vitro, where it behaves as a Michaelian enzyme. Conclusion. Extensive research is needed to confirm the allosteric mechanism of the vanilla β-glucosidase in vivo.


Corresponding author

* Correspondence and reprints


Hide All
[1] Cameron K.M., Recent advances in the systematic biology of vanilla and related orchids (Orchidaceae: subfamily Vanilloideae), in: Proc. Vanilla, First Int. Cong., Princeton, USA, Carol Stream, Allured Publ. Corp., USA, 2005, pp. 89–93.
[2] Havkin-Frenkel, D., French, J.C., Pak, F. Frenkel, C., Inside vanilla: Vanilla planifolia’s botany, curing options and future market prospects, Perfum. Flavour. 30 (2005) 3655.
[3] Odoux, E., Brillouet, J.-M., Anatomy, histochemistry and biochemistry of glucovanillin, oleoresin and mucilage accumulation sites in green mature vanilla pod (Vanilla planifolia; Orchidaceae): a comprehensive and critical reexamination, Fruits 64 (2009) 221241.
[4] Odoux, E., Escoute, J., Verdeil, J.-L. Brillouet, J.-M., Localization of β-glucosidase activity and glucovanillin in vanilla bean (Vanilla planifolia Andrews), Ann. Bot. 92 (2003) 437444.
[5] Odoux, E., Chauwin, A. Brillouet, J.-M., Purification and characterization of vanilla bean (Vanilla planifolia Andrews) β-D-glucosidase, J. Agric. Food Chem. 51 (2003) 31683173.
[6] Kanisawa, T Kanisawa, T., Flavor development in vanilla beans, Kouryou 180 (1993) 113123.
[7] Arana, F.E Arana, F.E., Action of a β-glucosidase in the curing of vanilla, Food Res. 288 (1943) 343351.
[8] Odoux, E Odoux, E., Changes in vanillin and glucovanillin concentrations during the various stages of the process traditionally used for curing Vanilla fragrans in Réunion, Fruits 55 (2000) 119125.
[9] Dignum, M.J.W., Kerler, J. Verpoorte, R., Vanilla curing under laboratory conditions, Food Chem. 79 (2002) 165171.
[10] Dignum, M.J.W., van der Heijden, R., Kerler, J., Winkel, C. Verpoorte, R., Identification of glucosides in green beans of Vanilla planifolia Andrews and kinetics of vanilla β-glucosidase, Food Chem. 85 (2004) 199205.
[11] Odoux, E., Escoute, J. Verdeil, J.-L., The relation between glucovanillin, β-glucosidase activity and cellular compartmentation during the senescence, freezing and traditional curing of vanilla beans, Ann. Appl. Biol. 149 (2006) 4352.
[12] Márquez, O. Waliszewski, K.N., The effect of thermal treatment on β-glucosidase inactivation in vanilla bean (Vanilla planifolia Andrews), Int. J. Food Sci. Technol. 43 (2008) 19931999.
[13] Roux P., Études morphologiques et anatomiques dans le genre Vanilla, in: Bouriquet G. (Ed.), Le vanillier et la vanille dans le monde, Lechevalier, Paris, France, 1954, pp. 44–92.
[14] French J.C., Development of vanilla-bearing placental trichomes, in: Proc. Vanilla, First Int. Cong., Princeton, USA, Carol Stream, Allured Publ. Corp., USA, 2005, pp. 71–77.
[15] Krishnakumar, V., Bindumol, G.P., Potty, S.N. Govindaraju, C., Processing of vanilla (Vanilla planifolia Andrews) beans – Influence of storing fresh beans, killing temperature and duration of killing on quality parameters, J. Spices Aromat. Crops 16 (2007) 3137.
[16] Hanum, T Hanum, T., Changes in vanillin and activity of β-glucosidase and oxidases during post harvest processing of vanilla beans (Vanilla planifolia), Bull. Teknol. Ind. Pagan 8 (1997) 4652.
[17] Márquez, O., Waliszewski, K.N. Oliart, R.M., Pardio V.T. Purification and characterization of cell-wall bound peroxidase from vanilla bean, Lebens. Wiss. u-Technol. 41 (2008) 13721379.
[18] Waliszewski, K., Márquez, O. Pardio, V.T., Quantification and characterization of polyphenol oxidase from vanilla bean, Food Chem. 117 (2009) 196203.
[19] Kanisawa T., Tokoro K., Kawahara S., Flavour development in the beans of Vanilla planifolia, in: Kurihara K., Suzuki N., Ogawa H. (Eds.), Olfaction taste XI, Proc. Int. Symp., Springer, Tokyo, Japan, 1994, pp. 268–270.
[20] Perez Silva A., Contribution à l’étude de la genèse des composés d’arôme au cours du procédé mexicain de transformation de la vanille (Vanilla planifolia Jackson), PhD Thesis, Univ. Montpellier II, France, 2006.
[21] Yonetani, T., Park, S., Tsuneshige, A., Imai, K. Kanaori, K., Global allostery model of hemoglobin – Modulation of O2 affinity, cooperativity, and Bohr effect by heterotropic allosteric effectors, J. Biol. Chem. 277 (2002) 3450834520.
[22] Perez-Silva, A., Odoux, E., Brat, P., Ribeyre, F., Rodriguez-Jimenes, G., Robles-Olvera, V., Garcia-Alvarado, M.A. Günata, Z., GC-MS and GC-olfactometry analysis of aroma compounds in a representative organic aroma extract from cured vanilla (Vanilla planifolia G. Jackson) beans, Food Chem. 99 (2006) 728735.
[23] Roberts, J.K.M., Ray, P.M., Wade-Jardetzky, N. Jardetzky, O., Estimation of cytoplasmic and vacuolar pH in higher plants by 31P NMR, Nat. 283 (1980) 870872.
[24] Mariezcurrena, M.D., Zavaleta, H.A., Waliszewski, K.N. Sanchez, V., The effect of killing conditions on the structural changes in vanilla (Vanilla planifolia Andrews) pods during the curing process, Int. J. Food Sci. Technol. 43 (2008) 14521457.
[25] Ranadive A.S., Szkutnica K., Guerrera J.G., Frenkel C., Vanillin biosynthesis in vanilla beans, in: Proc. IX Int. Cong. Ess. Oils, Singapore, 1983, pp. 147–154.


In vivo kinetics of β-glucosidase towards glucovanillin and related phenolic glucosides in heat-treated vanilla pod (Vanilla planifolia, Orchidaceae)

  • Jean-Marc Brillouet (a1) (a2) and Éric Odoux (a1)


Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed