Introduction. The traditional curing of vanilla pods includes “killing” andsweating 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 theirnon-aromatic glucosides is due to the action of an endogenous β-glucosidase, itsin vivo kinetics remained unknown. Materials and methods.Mature green vanilla pods were pretreated for 2 h at 50 ℃, 55 ℃ and 60 ℃, then stored for118 days at 27 ℃. Phenolic glucosides and their aglycons were extracted at regularintervals during the storage period and analyzed by HPLC. Results anddiscussion. All phenolic β-glucosides were slowly hydrolyzed during the storageperiod with production of vanillin, p-hydroxybenzaldehyde, vanillic acid,and other unknown aglycons. Most of the β-glucosidase was heat-denatured by thepretreatment, and analysis of its kinetic parameters showed that it adopts, invivo, an allosteric mode of functioning with a lower affinity for glucovanillinthan in vitro, where it behaves as a Michaelian enzyme.Conclusion. Extensive research is needed to confirm the allostericmechanism of the vanilla β-glucosidase in vivo.