Multienzyme engineering in the cellar: Versatile biotechnological potential of pectolytic-based clarification enzymes in rosé winemaking

Commercial pectolytic enzymes are widely used in winemaking for must clarification, but their side activities remain scarcely explored. The present study evaluated three commercial clarification enzymes (E1, E2, E3) for their collateral activities and their technological impact in both laboratory-scale and pilot-scale rosé winemaking. Enzymatic assays revealed that E1 exhibited the broadest and most intense accessory activity profile, including significant hemicellulolytic and glycosidase activities. Laboratory trials highlighted beneficial side effects, including increased yeast assimilable nitrogen and decreased calcium ions, an impact of potential interest for calcium tartrate stabilization. Pilot-scale rosé winemaking with Bombino nero and Primitivo grapes demonstrated that clarification enzymes can significantly modulate rosé wine composition, affecting colour indices and volatile profiles in a cultivar-dependent manner. Commercial pectolytic preparations can be considered versatile enzymatic cocktails that can be tailored to grape properties to optimise wine complexity and stability, moving beyond basic clarification toward multienzyme engineering. • Pectolytic enzymes were studied for rosé must clarification impact. • Enzymes show side activities (like -glycosidases) active at wine pH. • Clarification enzymes increase YAN and reduce calcium ions in must. • Enzyme impact on color and volatiles is cultivar-specific (Bombino/Primitivo).