Chiappini Viviana, Casbarra Debora, Astolfi Maria Luisa, Girelli Anna Maria
Department of Chemistry, Sapienza University of Rome, P.le A. Moro 5, Rome 00185, Italy.
Department of Chemistry, Sapienza University of Rome, P.le A. Moro 5, Rome 00185, Italy.
J Biotechnol. 2024 Sep 10;392:118-127. doi: 10.1016/j.jbiotec.2024.07.001. Epub 2024 Jul 3.
This study aimed at Candida rugosa lipase immobilization on a low-cost and readily available support. Among agro-industrial crops, hemp tea waste was chosen as the carrier because it provides higher immobilization performance than hemp flower and leaf wastes. Support characterization by ATR-FTIR, SEM and elemental analysis and the optimization of the adsorption immobilization process were performed. The lipase adsorption immobilization was obtained by soaking the support with hexane under mild agitation for 2 h and a successively incubating the enzyme for 1 h at room temperature without removing the solvent. The esterification of oleic acid with n-decanol was tested in a solvent-free system by studying some parameters that influence the reaction, such as the substrates molar ratio, the lipase activity/oleic acid ratio, reaction temperature and the presence/absence of molecular sieves. The biocatalyst showed the ability to bring the esterification reaction to equilibrium under 60 min and good reusability (maintaining 60 % of its original activity after three successive esterification reactions) but low conversion (21 %) at the optimized conditions (40 °C, 1:2 substrates molar ratio, 0.56 lipase/oleic acid ratio, without sieves). Comparing the results with those obtained by free lipase form at the same activity (1 U) and experimental conditions, slightly higher conversion (%) appeared for the free lipase. All this highlighted that probably the source of lipase for its carbohydrate-binding pocket and lid structure affected the esterification of oleic acid but certainly, the immobilization didn't induce any lipase conformational change also allowing the reuse of the catalytic material.
本研究旨在将皱褶假丝酵母脂肪酶固定在低成本且易于获得的载体上。在农业工业作物中,选择麻茶叶废料作为载体,因为它比麻花花叶废料具有更高的固定化性能。通过衰减全反射傅里叶变换红外光谱(ATR-FTIR)、扫描电子显微镜(SEM)和元素分析对载体进行了表征,并对吸附固定化过程进行了优化。脂肪酶的吸附固定化是通过将载体在温和搅拌下用己烷浸泡2小时,然后在室温下连续孵育酶1小时,且不除去溶剂来实现的。在无溶剂体系中,通过研究一些影响反应的参数,如底物摩尔比、脂肪酶活性/油酸比、反应温度以及分子筛的存在与否,对油酸与正癸醇的酯化反应进行了测试。该生物催化剂在60分钟内能够使酯化反应达到平衡,并且具有良好的可重复使用性(在连续三次酯化反应后仍保持其原始活性的60%),但在优化条件(40℃、底物摩尔比1:2、脂肪酶/油酸比0.56、无分子筛)下转化率较低(21%)。将结果与在相同活性(1 U)和实验条件下游离脂肪酶的结果进行比较,游离脂肪酶的转化率略高。所有这些都表明,可能是脂肪酶的碳水化合物结合口袋和盖子结构的来源影响了油酸的酯化反应,但可以肯定的是,固定化并没有引起任何脂肪酶构象变化,还允许催化材料的重复使用。