Multi-enzyme co-immobilized nano-assemblies: Bringing enzymes together for expanding bio-catalysis scope to meet biotechnological challenges.

Tailoring metal-organic frameworks-based nanozymes for bacterial theranostics.

Enzymes are the highly efficient biocatalyst in modern biotechnological industries. Due to the fragile property exposed to the external stimulus, the application of enzymes is highly limited. The immobilized enzyme by polymer has become a research hotspot to empower enzymes with more extraordinary properties and broader usage. Compared with free enzyme, polymer immobilized enzymes improve thermal and operational stability in harsh environments, such as extreme pH, temperature and concentration. Furthermore, good reusability is also highly expected. The first part of this study reviews the three primary immobilization methods: physical adsorption, covalent binding and entrapment, with their advantages and drawbacks. The second part of this paper includes some polymer applications and their derivatives in the immobilization of enzymes.

Immobilization of Enzymes by Polymeric Materials

Although capsule-like materials as host carriers for enzyme encapsulation have been a hot topic in recent years, creating an ideal microenvironment for enhanced enzymatic performance is still a formidable challenge. Herein, we created a template-free method to in situ encapsulate natural enzymes in hollow covalent organic framework (COF) capsules at room temperature. The COF crystallites migrated from the inner core and self-assembled at the outside walls during the inside-out Ostwald ripening process, retaining the enzymes in the cavity. The adjustable hollow structure of the enzyme@COF capsule allowed the basic vibration of the enzyme to maintain a certain degree of freedom, thus significantly enhancing the enzymatic bioactivity. The hollow enzyme@COF capsule has large mesoporous tunnels allowing the efficient transport. In addition, the enzyme encapsulated in the capsule showed superior activity and ultrahigh stability under various extreme conditions that may lead to enzyme inactivation, such as high temperature, organic solvents, chelates, and the denaturing agent. Finally, the prepared hollow GOx@COF capsule was used for electrochemical sensing of glucose in human serum, and the electrochemical sensor exhibited high selectivity and satisfactory test results. This research not only provides a new way for COFs to encapsulate enzymes but also has potential applications in biocatalysis and biosensing, making artificial organelles possible.

Template-Free In Situ Encapsulation of Enzymes in Hollow Covalent Organic Framework Capsules for the Electrochemical Analysis of Biomarkers.

Co-delivery of enzymes and photosensitizers via metal-phenolic network capsules for enhanced photodynamic therapy

Inspired by the unique biological microenvironments of eukaryotic cells, hollow capsules are promising to immobilize enzymes due to their advantages for physical protection and improved activity of enzymes. Herein, we report a facile method to fabricate silica (SiO2) capsules using zeolitic imidazole framework-8 nanoparticles (ZIF-8 NPs) as templates for enzyme immobilization and catalysis. Enzyme-encapsulated SiO2 capsules are obtained by encapsulation of enzymes in ZIF-8 NPs and subsequent coating of silica layers, followed by the removal of templates in a mild condition (i.e., ethylenediaminetetraacetic acid (EDTA) solution). The enzyme (i.e., horseradish peroxidase, HRP) activity in SiO2 capsules is improved more than 15 times compared to that of enzyme-loaded ZIF-8 NPs. Enzymes in SiO2 capsules maintain a high relative activity after being subjected to high temperature, enzymolysis, and recycling compared to free enzymes. In addition, multienzymes (e.g., glucose oxidase and HRP) can also be coencapsulated within SiO2 capsules to show a reaction with a high cascade catalytic efficacy. This work provides a versatile strategy for enzyme immobilization and protection with potential applications in biocatalysis.

Silica Capsules Templated from Metal–Organic Frameworks for Enzyme Immobilization and Catalysis

Understanding the impact of the physicochemical properties of nanoparticles (NPs) on cellular uptake is important to design optimal drug-delivery nanocarriers. Therein, the influence of NP elasticity on bio-nano-interactions remains elusive due to the complexity of factors affecting cellular uptake. Herein, we synthesized SiO2 capsules with tunable elasticity using metal-organic frameworks as templates to investigate their interactions with cells. Young's moduli of the resultant water-filled SiO2 capsules with identical size, shape, composition, and surface charge can be controlled from 3.8 MPa to 4.7 GPa via the variation of capsule shell thickness. As a result, increased elasticity of SiO2 capsules results in higher cellular uptake. Stiff SiO2 capsules have almost 9 times as much cellular uptake as the soft ones. In addition, the elasticity of SiO2 capsules influences cellular uptake pathways, where the clathrin-mediated pathway is preferred for stiff capsules while the uptake of the soft capsules is mostly mediated by a caveolae-dependent pathway. This work confirms the important role of NP elasticity in nonspecific cell interactions, which can provide a foundational understanding for engineering drug-delivery nanocarriers.

Xuebin Ma

Papers

Silica Capsules Templated from Metal–Organic Frameworks for Enzyme Immobilization and Catalysis

Effect of Elasticity of Silica Capsules on Cellular Uptake.