Li Hongxia, Wang Herui, Yang Xin, Zheng Hongru, Sun Chunyan, Ye Haiqing, Li Chenzhong, Yan Xu
Department of Food Quality and Safety, College of Food Science and Engineering, Jilin University, Changchun, 130062, China; College of Electronic Science and Engineering, Jilin University, Changchun, 130012, China.
Department of Food Quality and Safety, College of Food Science and Engineering, Jilin University, Changchun, 130062, China.
Biosens Bioelectron. 2025 Feb 15;270:116939. doi: 10.1016/j.bios.2024.116939. Epub 2024 Nov 14.
The development of a suitable mimetic scaffolds for maintaining high activity and stability of co-immobilized multi-enzymes is a key challenge in biotechnology. Herein, we achieved the regular distribution of cascade enzymes through spatially controlled hierarchical loading into protein-inorganic hybrid nanoflowers using a mild biomineralization technique. The comprehensive understanding of sequential regulation in constructing controlled nanoarchitecture enables to combine a continuous reaction and achieve tailoring catalysis for biomimetic application. The ordered-assembled cascade enzymes showed stronger bioactivity in comparison with the disordered format or inappropriate loading format. The stability of the enzyme is incrementally improved by an efficient dual-enhanced mode of immobilizing the free enzyme into hybrid nanoflowers and encapsulating it in a hydrogel system, addressing the inherent fragility of natural enzymes. Benefiting from the structural integration, a protein-inorganic hybrid nanoflowers-embedded hydrogel sensor is constructed for on-site detecting NO with a detection limit of 5.08 μM. This work showcases a convenient approach for the efficient design of the ideal cascade biocatalysts, and supports the development of portable devices for practical application.
开发一种合适的模拟支架以维持共固定化多酶的高活性和稳定性是生物技术中的一项关键挑战。在此，我们通过使用温和的生物矿化技术将级联酶空间控制地分级负载到蛋白质-无机杂化纳米花中，实现了级联酶的规则分布。对构建可控纳米结构中顺序调控的全面理解使得能够结合连续反应并实现用于仿生应用的定制催化。与无序形式或不合适的负载形式相比，有序组装的级联酶表现出更强的生物活性。通过将游离酶固定到杂化纳米花中并将其封装在水凝胶系统中的高效双增强模式，逐步提高了酶的稳定性，解决了天然酶固有的脆弱性问题。受益于结构整合，构建了一种嵌入蛋白质-无机杂化纳米花的水凝胶传感器，用于现场检测NO，检测限为5.08 μM。这项工作展示了一种有效设计理想级联生物催化剂的便捷方法，并支持了实际应用中便携式设备的开发。
