Amorphous metal-organic frameworks on PtCu hydrogels: Enzyme immobilization platform with boosted activity and stability for sensitive biosensing

Cell-free enzymatic catalysis (CFEC) is emerging biotechnology that simulates biological transformations without living cells. However, the high cost of separation and preparation of the enzyme has hindered the practical application of the CFEC. Enzyme immobilization technologies using solid supports to stabilize enzymes have been regarded as an efficient strategy to address this issue. Nevertheless, the activity and stability of the immobilized enzymes are still crucial challenges for working in vitro. Herein, an enzyme immobilization platform is developed by using PtCu hydrogels coated with amorphous metallic-organic frameworks (MOFs) as multifunctional carriers to encapsulate horseradish peroxidase (HRP). Specifically, PtCu hydrogels acting as a "reservoir of metal ions" can interact with the immobilized enzyme and facilitate electron transfer, leading to the boosted enzyme catalytic performances. Furthermore, amorphous MOFs on the surface of PtCu hydrogels serve as an "armor" to protect the internal enzymes from various perturbation environments. The resultant enzyme immobilization platform (PtCu@HRP@ZIF-8) not only shows an approximately 2.4-fold enhanced activity compared with free enzyme but also exhibits improved stability against harsh conditions. The PtCu@HRP@ZIF-8-based biosensor is constructed for sensitive sensing of organophosphorus pesticides (OPs). The proposed biosensor exhibits a favorable linear relationship with the concentration of paraoxon-ethyl from 6 to 800 ng/mL, with a low detection limit of 1.8 ng/mL. This work reveals the promising potential of our proposed enzyme immobilization platform in practical applications.