Sustainable Enzymatic Production of Bioactive Compounds: From Designing to Bioavailability

This study evaluates the feasibility and effectiveness of using immobilized pectinase enzymes for juice processing to reduce cloudiness while preserving nutritional and bioactive properties. The research is driven by the increasing demand for innovative food products that offer enhanced functionality and health benefits. It focuses on the development and application of immobilized biocatalysts in bioprocessing, specifically using pectinase encapsulated in a sol–gel matrix. Reaction parameters for the interaction between immobilized pectinase and its primary substrate, pectin, were optimized through systematic experimentation. Optimal conditions were established, achieving enhanced enzyme activity and stability with 0.15 g of lens-shaped capsules containing 10.0 mg/mL pectinase in 24-well microplates as microreactors. Kinetic studies indicated improved substrate affinity after immobilization (Km = 0.115 mg/mL), particularly when magnetized (Kmi = 0.041 mg/mL). Operational stability and reusability assessments demonstrated potential for extended use with magnetized pectinase capsules retaining higher residual activity after a fourth reuse cycle (155% > 75%). The application of immobilized pectinase in processing peach nectar successfully reduced cloudiness and increased the release of bioactive compounds, enhancing antioxidant and anti-inflammatory activities, as evaluated by the DPPH (2,2-diphenyl-1-picrylhydrazyl) assay and the albumin method, respectively. In vitro digestion studies revealed dynamic activity profile changes, highlighting the impact of juice bioprocessing on bioavailability.