CN-121991940-A - Enzyme-loaded porous gel capable of being rapidly reconstituted, preparation method and flow microreactor

CN121991940ACN 121991940 ACN121991940 ACN 121991940ACN-121991940-A

Abstract

The invention relates to a rapid-reconfiguration enzyme-carrying porous gel, a preparation method and a flow microreactor, wherein amphiphilic nano particles are used as a stabilizer, and a porous gel structure with uniform and adjustable pore diameter is formed by introducing bubbles into a high polymer solution and inducing polymerization and crosslinking, and the amphiphilic nano particles can be adsorbed on a gas-liquid interface to stabilize the bubbles and can carry enzyme molecules on the surface of the amphiphilic nano particles to form a porous reaction medium with catalytic activity. The obtained porous gel has good mechanical strength, controllable pore diameter and porosity, and can realize rapid reconstruction of a porous structure, thereby widening the applicable scene of the microreactor and remarkably prolonging the effective service period of enzyme when the microreactor is blocked.

Inventors

ZHANG SHAOBIN
LI PEIHAO
YU ZIYI
LI JUAN

Assignees

南京工业大学

Dates

Publication Date: 20260508
Application Date: 20251231

Claims (10)

1. The enzyme-carrying porous gel capable of being rapidly reconstituted is characterized by comprising a solid gel network and amphiphilic nano-particles with the surfaces loaded with enzymes, wherein the solid gel network forms a main body structure of the enzyme-carrying porous gel, and the amphiphilic nano-particles are distributed in the solid gel network; The solid gel network comprises a polymer material which can be degraded or can realize reversible sol-gel conversion, so that the solid gel network can be degraded or converted into a solution state under a specific action; the amphiphilic nanoparticles comprise a material that is insensitive to the specific effects, such that the amphiphilic nanoparticles are retained during the transition of the solid gel network into solution and used to reconstruct other enzyme-loaded porous gels.
2. The enzyme-loaded porous gel according to claim 1, wherein the enzyme is loaded on the surface of the amphiphilic nano particles in a physical adsorption or chemical coupling manner and distributed on the pore walls of the solid gel network along with the amphiphilic nano particles, thereby realizing an enzyme catalytic function.
3. The enzyme-loaded porous gel according to claim 1, wherein the pores of the solid gel network are prepared according to a bubble template method, the amphiphilic nanoparticles are used for stabilizing a bubble structure, and the pore size distribution and the porosity of the solid gel network are controlled by regulating the concentration of the amphiphilic nanoparticles.
4. A flow microreactor based on a rapidly reconfigurable enzyme-loaded porous gel, the flow microreactor comprising: A microreactor body including a through channel through which a reactant flows; a porous enzyme-loaded gel according to any one of claims 1-3, disposed within the through-channels for enzyme catalysis of the reactants.
5. A method for preparing a rapidly reconfigurable enzyme-loaded porous gel according to any one of claims 1 to 3, comprising the steps of: (1) Fixing one or more enzymes on the surface of the amphiphilic nano-particles to obtain the amphiphilic nano-particles with the surfaces loaded with the enzymes; (2) Mixing amphiphilic nano particles with enzyme loaded on the surfaces with one or more precursor polymers, introducing bubbles through a bubble template method to form a compact bubble structure, and solidifying the polymer mixed solution to obtain the enzyme-loaded porous gel; Or alternatively (1) Mixing the amphiphilic nano particles with one or more precursor polymers, introducing bubbles through a bubble template method to form a compact bubble structure, and solidifying the polymer mixed solution to obtain porous gel; (2) And fixing one or more enzymes on the surfaces of amphiphilic nano particles on the pore walls of the porous gel to obtain the enzyme-carrying porous gel.
6. The method of manufacturing according to claim 5, further comprising the steps of: Degrading or colloidizing the enzyme-carrying porous gel through the specific action to remove a solid gel network in the enzyme-carrying porous gel, wherein the specific action is a treatment mode which is matched with the material property of the solid gel network and comprises enzyme response degradation, pH response degradation, temperature response gel-collosol conversion and the like; separating the amphiphilic nanoparticles loaded with enzymes on the surface from the solution obtained in the previous step by one or more methods of centrifugation, dialysis or filtration; And (3) remixing the amphiphilic nano particles with the enzyme loaded on the surfaces with one or more precursor polymers, introducing bubbles through a bubble template method to form a compact bubble structure, and solidifying the polymer mixed solution to obtain the reconstructed enzyme-loaded porous gel.
7. The preparation method of the nano-particles according to claim 5 or 6, wherein the amphiphilic nano-particles are organic or inorganic amphiphilic nano-particles, wherein the organic amphiphilic nano-particles comprise block self-assembled nano-particles, natural polymer-based amphiphilic nano-particles and the like, and the inorganic amphiphilic nano-particles comprise silicon-based amphiphilic nano-particles, metal/metal oxide-based amphiphilic nano-particles, carbon-based amphiphilic nano-particles and the like.
8. The method according to claim 5 or 6, wherein the precursor polymer is one or more of a natural polymer group and its modified derivative, a synthetic polymer group, and a composite polymer group.
9. The method of claim 5 or 6, wherein the immobilization of one or more of the enzymes to the amphiphilic nanoparticles is achieved by means of physical adsorption or chemical coupling.
10. The method according to claim 5 or 6, wherein the bubble template method comprises forming bubbles in the polymer mixed solution by direct bubbling, microfluidic technology or high-speed shearing by a homogenizer, and the like, patterning the polymer mixed solution containing bubbles to form a specific geometric structure, and then solidifying.

Description

Enzyme-loaded porous gel capable of being rapidly reconstituted, preparation method and flow microreactor Technical Field The invention relates to the technical field of enzyme catalysis, in particular to a rapid-reconfigurable enzyme-carrying porous gel, a preparation method and a flow microreactor. Background The enzyme catalysis technology has important application in the fields of fine chemical engineering, green chemistry, biological manufacturing and the like. In order to achieve the continuity and the high efficiency of the enzyme catalysis process, a flow microreactor constructed based on porous gel materials has been receiving a great deal of attention. Such porous gels typically form a three-dimensional porous network by physical or chemical cross-linking and immobilize the enzyme molecules on the pore wall surface to build a stable enzyme catalytic system. In the prior art, to achieve stable loading of enzymes, it is often necessary to introduce polymers with reactive functional groups to form porous gel structures that can immobilize the enzyme. However, in the actual immobilization process, a large number of functional groups in the gel main body cannot participate in enzyme immobilization, and only a small number of functional groups on the surface of the pore wall can be effectively utilized, so that the enzyme loading form is limited and the loading amount is insufficient, thereby influencing the catalytic efficiency. In addition, the porous gel flow microreactor is easy to block under the continuous flow condition, the enzymes fixed on the pore wall are difficult to realize effective recovery after blocking, and the enzymes can only be discarded along with the whole gel, so that enzyme resource waste is caused, and the running cost is obviously increased. Further, the pore size, porosity and channel network of conventional porous gels are essentially defined after solidification and formation, and are difficult to readjust according to the reaction requirements, resulting in such porous gels being only suitable for use in reactors having specific flow rate ranges, substrate diffusion conditions and enzyme systems. When the switching between different reaction scenes is needed (such as the switching from a small molecular substrate to a large molecular substrate or the operation between different flow rates and pressure ranges), the existing porous gel structure cannot be quickly reconstructed, and the multi-scene adaptation requirement of continuous flow reaction is difficult to meet. Disclosure of Invention The invention aims to solve the problems that the existing porous gel-based microreactor is limited in enzyme load form and insufficient in load capacity, enzymes are difficult to recycle when channels are blocked, the microreactor structure is difficult to regulate and control as required, and the like, and provides an enzyme-loaded porous gel capable of being rapidly reconstructed, a preparation method and a flowing microreactor so as to realize multiple advantages of high-efficiency enzyme load, rapid recovery and adjustable reactor structure. As one aspect of the present invention, there is provided a rapid-reconfigurable enzyme-carrying porous gel comprising a solid gel network and amphiphilic nanoparticles carrying enzymes on the surface, the solid gel network constituting the main structure of the enzyme-carrying porous gel, the amphiphilic nanoparticles being distributed in the solid gel network; wherein the solid gel network comprises a polymer material which can be degraded or can realize reversible sol-gel conversion, so that the solid gel network can be degraded or converted into a solution under a specific action; the amphiphilic nanoparticles comprise a material that is insensitive to the specific effects, such that the amphiphilic nanoparticles are retained during the transition of the solid gel network into solution and used to reconstruct other enzyme-loaded porous gels. Optionally, with reference to any one of the foregoing aspects, in another implementation manner of this aspect, the enzyme is supported on a surface of the amphiphilic nanoparticle by a physical adsorption or chemical coupling manner, and is distributed on a pore wall of the solid gel network along with the amphiphilic nanoparticle, so as to implement an enzyme catalytic function. Optionally, in combination with any of the above aspects, in another implementation of the present aspect, the enzyme species includes one or more, thereby implementing a single or multiple enzyme coupled catalytic function. Optionally, with reference to any one of the foregoing aspects, in another implementation manner of this aspect, the pores of the solid gel network are prepared according to a bubble template method, the amphiphilic nanoparticles are used for stabilizing a bubble structure, and the pore size distribution and the porosity of the solid gel network are controlled by regulating the concentration of the amphip