Martínez-Baquero Raúl, Martínez-Tomé María José, Gómez Javier, Esquembre Rocío, Mateo C Reyes
Instituto de Investigación, Desarrollo e Innovación en Biotecnología Sanitaria de Elche (IDiBE), Universidad Miguel Hernández (UMH), 03202 Elche, Spain.
Biosensors (Basel). 2025 Sep 17;15(9):615. doi: 10.3390/bios15090615.
Eutectogels are advanced gel-based systems that integrate deep eutectic solvents (DES) into polymer networks. In this study, we report the first detailed characterization of an enzyme-containing eutectogel, representing a significant step toward advanced biosensing and biocatalytic applications. Specifically, we have incorporated pancreatic lipase, one of the main target enzymes in the treatment of obesity, in eutectogels via UV-induced radical polymerization of suitable precursors in appropriate DESs. Prior to immobilization, the enzyme was solubilized in selected DESs and its activity and conformational stability were evaluated using colorimetry and intrinsic fluorescence. Combinations of choline chloride/glycerol and tetramethylammonium chloride/glycerol were shown to be effective media for preserving and enhancing enzymatic function and conformational stability. The enzyme immersed in eutectogel exhibited high structural integrity and excellent thermal stability, maintaining its activity over several weeks. The ability of this new material to screen enzyme inhibitors was assessed using orlistat, a well-established anti-obesity agent. The results demonstrated clear detection of the drug's inhibitory effect, even at nanomolar concentrations, highlighting the material's potential as a screening platform for novel inhibitors with prospective anti-obesity activity. Furthermore, the device proved effective in quantifying drug presence, offering a promising and highly sensitive tool for pharmaceutical quality control applications.
低共熔凝胶是一种先进的基于凝胶的体系，它将低共熔溶剂（DES）整合到聚合物网络中。在本研究中，我们首次详细表征了一种含酶低共熔凝胶，这是朝着先进的生物传感和生物催化应用迈出的重要一步。具体而言，我们通过在合适的DES中对合适的前体进行紫外诱导自由基聚合，将肥胖治疗中的主要靶标酶之一胰脂肪酶纳入低共熔凝胶中。在固定化之前，将酶溶解在选定的DES中，并使用比色法和固有荧光评估其活性和构象稳定性。结果表明，氯化胆碱/甘油和四甲基氯化铵/甘油的组合是保存和增强酶功能及构象稳定性的有效介质。浸入低共熔凝胶中的酶表现出高度的结构完整性和出色的热稳定性，其活性在数周内得以维持。使用已确立的抗肥胖药物奥利司他评估了这种新材料筛选酶抑制剂的能力。结果表明，即使在纳摩尔浓度下，也能清晰检测到该药物的抑制作用，突出了该材料作为具有潜在抗肥胖活性的新型抑制剂筛选平台的潜力。此外，该装置在定量药物存在方面被证明是有效的，为药物质量控制应用提供了一种有前景且高度灵敏的工具。
