Spatially ordered immobilization of cascade enzymes for the construction of a robust colorimetric hydrogel sensor

Hongxia Li; Herui Wang; Xin Yang; Hongru Zheng; Chunyan Sun; Haiqing Ye; Chenzhong Li; Xu Yan

doi:10.1016/j.bios.2024.116939

Spatially ordered immobilization of cascade enzymes for the construction of a robust colorimetric hydrogel sensor

Biosens Bioelectron. 2024 Nov 14:270:116939. doi: 10.1016/j.bios.2024.116939. Online ahead of print.

Authors

Hongxia Li¹, Herui Wang², Xin Yang², Hongru Zheng³, Chunyan Sun², Haiqing Ye⁴, Chenzhong Li⁵, Xu Yan⁶

Affiliations

¹ 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.
² Department of Food Quality and Safety, College of Food Science and Engineering, Jilin University, Changchun, 130062, China.
³ Jilin Province Product Quality Supervision and Inspection Institute, Changchun, 130103, Jilin, China.
⁴ Department of Food Quality and Safety, College of Food Science and Engineering, Jilin University, Changchun, 130062, China. Electronic address: yehq@jlu.edu.cn.
⁵ Biomedical Engineering Division, Chinese University of Hong Kong (Shenzhen), Shenzhen, 518172, China. Electronic address: lichenzhong@cuhk.edu.cn.
⁶ College of Electronic Science and Engineering, Jilin University, Changchun, 130012, China. Electronic address: yanx@jlu.edu.cn.

PMID: 39566331
DOI: 10.1016/j.bios.2024.116939

Abstract

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 NO₂^- 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.

Keywords: Biosensor; Cascade catalysis; Enzyme immobilization; Hydrogel; Spatial control.