Enzyme-assisted extraction in liquid-liquid two-phase aqueous system was applied for the first time in order to extract genipin from genipap. The effect of different commercial enzymes, their concentrations, and extraction parameters were investigated. Moreover, chitosan gels were prepared, crosslinked with glutaraldehyde or genipin and characterized by their textural and rheological properties. The crosslinked chitosan was used as support for the immobilization of model β-galactosidases. Among the different commercial enzymes tested for extraction, Celluclast 10% (36 °C and pH 3.7) provided an extraction of 196 mg.g of genipin. Chitosan gels crosslinked with genipin 0.5% showed better textural and similar rheological properties when compared to the chitosan crosslinked with glutaraldehyde 3%. The percentage of lactose hydrolysis by the immobilized K. lactis β-galactosidase using genipin as a crosslinker was 87%. Thus, the genipin obtained in this work proved to be an excellent alternative to the use of glutaraldehyde in chitosan crosslinking applications.

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In this work, we studied the covalent immobilization of α‐acetolactate decarboxylase (ALDC) on glutaraldehyde activated chitosan beads for application in beers to prevent off‐flavor. The immobilized preparation was characterized regarding yield, efficiency, activity recovery, thermal and pH stabilities, and operational stability in beer. The concentration of activating agent was evaluated and at the best condition an immobilization yield of 84% and activity recovery of 9.1% were verified. Optimal pH was 6.0 for free and immobilized enzymes, however, the immobilized enzyme showed higher relative activity at the tested pH, with emphasis on the pH range from 5.0 to 6.5. Besides, immobilized enzyme showed stabilization factor of almost sevenfold at 60°C compared to free form. The immobilized biocatalyst was tested in repeated batches in beer for α‐acetolactate conversion to acetoin, and after 12 cycles the enzyme presented 80% of its initial activity. To date, this is the most stable immobilized preparation of ALDC reported.

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