Lordelo Nascimento Luciana, Pita Bruna Louise de Moura, Rodrigues César de Almeida, Dos Santos Paulo Natan Alves, Almeida Yslaine Andrade de, Ferreira Larissa da Silveira, de Oliveira Maira Lima, de Almeida Lorena Santos, Soares Cleide Maria Faria, de Souza Dias Fabio, Fricks Alini Tinoco
Programa de Pós-Graduação em Ciência de Alimentos, Universidade Federal da Bahia, Salvador 40170-115, BA, Brazil.
Departamento de Análises Bromatológicas, Faculdade de Farmácia, Universidade Federal da Bahia, Salvador 40170-115, BA, Brazil.
Molecules. 2025 Jul 30;30(15):3207. doi: 10.3390/molecules30153207.
This study reports the development of a sustainable biocatalyst system for free fatty acid (FFA) production from cocoa bean shell (CBS) oil using lipase (). 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·min (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; ΔH‡ = 29.7 kJ mol), 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.
本研究报道了一种可持续生物催化剂系统的开发，该系统使用脂肪酶从可可豆壳（CBS）油中生产游离脂肪酸（FFA）。CBS被探索用作载体材料和反应底物。通过物理吸附或共价结合，使用有机（CBS）、无机（二氧化硅）和混合（CBS-二氧化硅）载体制备了六种固定化体系。其中，共价固定在CBS上的酶（ORG-CB）表现出最平衡的性能，催化效率（Ke）为0.063 mM·min（游离酶的18.6%），具有广泛的pH-温度耐受性，并且在八个重复使用循环后活性保留超过50%。热力学分析证实ORG-CB的耐热性增强（Ed = 32.3 kJ/mol；ΔH‡ = 29.7 kJ/mol），而动力学评估表明，在长时间加热下，其热失活比游离酶更快。在应用试验中，ORG-CB从CBS油中实现了60.1%的FFA转化率，优于游离酶（49.9%）。这些发现验证了CBS作为酶固定化和农业工业废弃物增值的双功能材料。结果还强化了固定化化学和载体组成对生物催化剂操作和热性能的影响，有助于推进基于酶的加工中的绿色化学策略。
