RESUMEN
This study reports the immobilization of a ß-CGTase on glutaraldehyde pre-activated silica and its use to production of cyclodextrins in batch and continuous reactions. We were able to modulate the cyclodextrin production (α-, ß- and γ-CD) by immobilization and changing the reaction conditions. In batch reactions, the immobilized enzyme reached to maximum productions of 4.9mgmL-1 of α-CD, 3.6mgmL-1 of ß-CD and 3.5mgmL-1 of γ-CD at different conditions of temperature, pH and reaction time. In continuous reactor, varying the residence time and pH it was possible to produce at pH 4.0 and 141min of residence time preferentially γ-CD (0.75 and 3.36mgmL-1 of α- and γ-CD, respectively), or at pH 8.0 and 4.81min α- and ß-CDs (3.44 and 3.51mgmL-1).
Asunto(s)
Enzimas Inmovilizadas/química , Glucosiltransferasas/química , gamma-Ciclodextrinas/síntesis química , Concentración de Iones de HidrógenoRESUMEN
Cyclodextrin glycosyltransferase (CGTase) from Thermoanaerobacter sp. was covalently immobilized on glutaraldehyde-activated chitosan spheres and used in a packed bed reactor to investigate the continuous production of ß-cyclodextrin (ß-CD). The optimum temperatures were 75 °C and 85 °C at pH 6.0, respectively for free and immobilized CGTase, and the optimum pH (5.0) was the same for both at 60 °C. In the reactor, the effects of flow rate and substrate concentration in the ß-CD production were evaluated. The optimum substrate concentration was 4% (w/v), maximizing the ß-CD production (1.32 g/L) in a flow rate of 3 mL/min. In addition, the biocatalyst had good operational stability at 60 °C, maintaining 61% of its initial activity after 100 cycles of batch and 100% after 100 h of continuous use. These results suggest the possibility of using this immobilized biocatalyst in continuous production of CDs.