CN-121975784-A - Hemoglobin-based micro-nano reactor and preparation method and application thereof

Abstract

The invention discloses a hemoglobin-based micro-nano reactor, a preparation method and application thereof, belongs to the technical field of biochemistry, and particularly relates to a hemoglobin-based micro-nano reactor which is formed by self-assembly of a hemoglobin-polylactic acid conjugate. The hemoglobin-based micro-nano reactor has a space structure similar to cells, has multiple functions of a drug delivery system and the micro-nano reactor, utilizes hemoglobin biomolecules in the hemoglobin-based micro-reactor to catalyze atom transfer radical polymerization reaction, can synthesize polymers in living cells under the condition of no need of additional catalyst or genetic modification, has the advantages of simple synthesis, good biocompatibility, various catalytic polymers and the like, and is suitable for the fields of cell engineering, biological material preparation, fluorescent labeling, polymer production by a fermentation method and the like.

Inventors

• WANG LINA
• WANG YUPENG
• HU YUHONG
• LI RUI
• HAN YANAN
• CHENG HONGYANG

Assignees

• 河北师范大学

Dates

Publication Date

20260505

Application Date

20260126

Claims (10)

1. 1. The hemoglobin-based micro-nano reactor is characterized by being formed by self-assembly of a hemoglobin-polylactic acid conjugate.
2. 2. The hemoglobin-based micro-nano reactor according to claim 1, wherein the particle size of the hemoglobin-based micro-nano reactor is 170 nm-2 μm, and the hemoglobin-based micro-nano reactor is a hollow vesicle structure.
3. 3. The hemoglobin-based micro-nano reactor according to claim 1, wherein the hemoglobin-polylactic acid conjugate is prepared by covalent linkage of hemoglobin subunits and alkenyl-terminated polylactic acid.
4. 4. The hemoglobin-based micro-nano reactor according to claim 3, wherein the alkenyl-terminated polylactic acid has a molecular weight of 15000 or 28000.
5. 5. The method for preparing the hemoglobin-based micro-nano reactor according to any one of claims 1 to 4, comprising the steps of: 1) Preparing an alkenyl-terminated polylactic acid by atom transfer radical polymerization; 2) Covalently linking the hemoglobin and the alkenyl-terminated polylactic acid obtained in the step 1) to form a hemoglobin-polylactic acid conjugate; 3) Dispersing the hemoglobin-polylactic acid conjugate obtained in the step 2) in a solvent to obtain a system A; 4) Dispersing a polymerization active monomer and a polymerization initiator in water to obtain a system B; 5) Dropwise adding the system A obtained in the step 3) into the system B obtained in the step 4) to obtain a system C; 6) Dialyzing the system C obtained in the step 5) to obtain the hemoglobin-based micro-nano reactor.
6. 6. The method according to claim 5, wherein the polymerization-active monomer in the step 4) comprises one or more of acrylamide, N-isopropyl acrylamide, N-acryloylmorpholine, hydroxyethyl methacrylate, and hydroxypropyl acrylate.
7. 7. The method according to claim 5, wherein the polymerization initiator in step 4) comprises one or more of ethyl α -bromophenylacetate, 2-hydroxyethyl-2-bromoisobutyrate, α -bromophenylacetic acid, and brominated polyethylene glycol.
8. 8. The use of the hemoglobin-based micro-nano reactor according to any one of claims 1-4 in catalyzing atom transfer radical polymerization.
9. 9. The use of claim 8, wherein the hemoglobin-based micro-nano reactor has a critical aggregation concentration of 0.018-0.024 mg/mL.
10. 10. The method according to claim 8, wherein the hemoglobin-based micro-nano reactor reaction system catalyzes atom transfer radical polymerization reaction at a controllable temperature, air is introduced to terminate the reaction, and an organic solvent is used for extraction to obtain the polymer.

Description

Hemoglobin-based micro-nano reactor and preparation method and application thereof Technical Field The invention belongs to the technical field of biochemistry, and particularly relates to a hemoglobin-based micro-nano reactor, and a preparation method and application thereof. Background As a bioorthogonal technology, the intracellular polymerization can directly synthesize polymers in living cells, and provides a brand new approach for cell function modification, novel biological material development and synthetic biology research. The traditional intracellular polymerization method is mostly dependent on a photoinitiation system or an exogenous inorganic catalyst, and has the problems of harsh reaction conditions, poor biocompatibility, easiness in damaging cells and the like. In addition, existing methods often require the expression of specific enzymes as catalysts by genetic modification, which is complex and costly to operate. Hemoglobin is the natural protein with the largest content in organisms, and has the advantages of wide sources, easy extraction, adjustable functions and the like. Hemoglobin has a special structure containing iron and has potential catalytic activity. If the natural catalytic capability of the hemoglobin can be utilized to trigger the polymerization reaction, genetic modification and exogenous catalyst are not needed, the intracellular polymerization process is obviously simplified, and the biocompatibility is improved. How to efficiently deliver the monomers and the initiator used by the hemoglobin and the synthetic polymer into the cells, and construct a stable polymerization system in the cells to realize efficient and controllable polymerization reaction is a technical problem to be solved in the field. The micro-nano carrier has wide application prospect in the fields of drug delivery, biocatalysis, tissue engineering and the like. Among them, protein-based nanocarriers have become a research hotspot in recent years due to good biocompatibility, biodegradability and functional adjustability. Therefore, in the art, development of a hemoglobin-based micro-nano carrier capable of efficiently supporting monomers and initiators used for synthesizing polymers, and realizing efficient and controllable intracellular polymerization reaction as a micro-nano reactor after being delivered into cells is desired. Disclosure of Invention In order to solve the technical problems, the invention provides a hemoglobin-based micro-nano reactor, a preparation method and application thereof, wherein the hemoglobin-based micro-nano reactor has a cell-like space structure, has multiple functions of a drug delivery system and the micro-nano reactor, utilizes the polymerization reaction of hemoglobin biomolecules catalytic atoms in the hemoglobin-based micro-reactor to transfer free radicals, and can synthesize polymers in living cells under the condition of no need of additional catalyst or genetic modification, and the micro-nano reactor has the advantages of simple synthesis, good biocompatibility, various catalytic polymer types and the like, and is suitable for the fields of cell engineering, biological material preparation, fluorescent labeling, polymer production by a fermentation method and the like. In order to achieve the above object, the present invention provides a hemoglobin-based micro-nano reactor formed by self-assembly of a hemoglobin-polylactic acid conjugate. Preferably, the particle size of the hemoglobin-based micro-nano reactor is 170 nm-2 mu m, and the hemoglobin-based micro-nano reactor is of a hollow vesicle structure. Preferably, the hemoglobin-polylactic acid conjugate is prepared by covalently linking a hemoglobin subunit to an alkenyl-terminated polylactic acid. Further preferably, the molecular weight of the alkenyl-terminated polylactic acid is 15000 or 28000. The invention also provides a preparation method of the hemoglobin-based micro-nano reactor, which comprises the following steps of 1) preparing alkenyl-terminated polylactic acid through atom transfer radical polymerization, 2) forming a hemoglobin-polylactic acid conjugate by covalent connection of hemoglobin and the alkenyl-terminated polylactic acid obtained in the step 1), 3) dispersing the hemoglobin-polylactic acid conjugate obtained in the step 2) in a solvent to obtain a system A, 4) dispersing a polymerization active monomer and a polymerization initiator in water to obtain a system B, 5) dropwise adding the system A obtained in the step 3) into the system B obtained in the step 4) to obtain a system C, and 6) dialyzing the system C obtained in the step 5) to obtain the hemoglobin-based micro-nano reactor. Preferably, the polymerization-active monomer in step 4) comprises one or more of acrylamide, N-isopropyl acrylamide, N-acryloylmorpholine, hydroxyethyl methacrylate, and hydroxypropyl acrylate. Preferably, the polymerization initiator in step 4) comprises one or more of ethyl α -bromophenylacetate