Immobilization of β-Glucosidase and Its Application for Enhancement of Aroma Precursors in Muscat Wine

Enzyme immobilization is becoming more widely practised in biotechnology because of the advantages that this method brings. In this study, commercial β-glucosidase for aroma released in winemaking was immobilized on diverse supports (alginate–chitin, chitosan–chitin) by using different methods. It was found that the most appropriate matrix was chitosan by adsorption and reticulation. The optimal immobilization conditions were pH 3.5, immobilization time 120 min, and concentration of cross-linker glutaraldehyde 0.25 %. Stability of the immobilized enzymes was assessed, which revealed a number of advantages, such as a lower enzyme dose required for immobilization (367 times lower than the free enzyme dose recommended by the manufacturer), high stability over time, and reusability. In vitro studies of cellobiose and in vivo studies of wine and aroma precursors isolated from grape must yielded similar outcomes with respect to enzyme hydrolysis of free and immobilized proteins.