This study reports the development of a sustainable biocatalyst system for free fatty acid (FFA) production from cocoa bean shell (CBS) oil using Burkholderia cepacia lipase (BCL). CBS was explored as both a support material and a reaction substrate. Six immobilized systems were prepared using organic (CBS), inorganic (silica), and hybrid (CBS–silica) supports via physical adsorption or covalent binding. Among them, the covalently immobilized enzyme on CBS (ORG-CB) showed the most balanced performance, achieving a catalytic efficiency (Ke) of 0.063 mM−1·min−1 (18.6% of the free enzyme), broad pH–temperature tolerance, and over 50% activity retention after eight reuse cycles. Thermodynamic analysis confirmed enhanced thermal resistance for ORG-CB (Ed = 32.3 kJ mol−1; ΔH‡ = 29.7 kJ mol−1), while kinetic evaluation revealed that its thermal deactivation occurred faster than for the free enzyme under prolonged heating. In application trials, ORG-CB reached 60.1% FFA conversion from CBS oil, outperforming the free enzyme (49.9%). These findings validate CBS as a dual-function material for enzyme immobilization and valorization of agro-industrial waste. The results also reinforce the impact of immobilization chemistry and support composition on the operational and thermal performance of biocatalysts, contributing to the advancement of green chemistry strategies in enzyme-based processing.
This study aimed to evaluate the effects of high-intensity ultrasound (40 W/5 min), applied with and without mild heating (59 °C and 23 °C), and of pasteurization (63 °C/30 min), on the physicochemical, rheological, and microbiological parameters, as well as on ascorbate oxidase activity, total carotenoid content, phenolic compound profile, and antioxidant capacity of passion fruit (Passiflora edulis Sims.) pulps. Ultrasound processing induced changes in color (L*, a*, and b*), resulting in high ∆E values. Following ultrasound treatment, an increase in apparent viscosity at 100 s−1 was observed. Ultrasound also promoted partial inactivation of ascorbate oxidase and a significant reduction in mold and yeast counts. Moreover, the application of ultrasound without heating (US-20) promoted the retention of 55% of ascorbic acid after 63 days of storage. The condition with heating (US-60) led to an increase in catechin content in both bright red passion fruit pulp (173.96%) and yellow passion fruit pulp (5.89%), demonstrating a balance between the retention of bioactive compounds, microbial inactivation, and reduction in ascorbate oxidase activity. Therefore, these results highlight ultrasound as a non-thermal and sustainable technology capable of extending shelf life, maximizing the preservation of bioactive compounds, and enhancing the functional properties of fruit pulps.