CN110585167A - Silk fibroin microcapsule loaded with horseradish peroxidase and preparation method thereof - Google Patents
Silk fibroin microcapsule loaded with horseradish peroxidase and preparation method thereof Download PDFInfo
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- 108010022355 Fibroins Proteins 0.000 title claims abstract description 53
- 239000003094 microcapsule Substances 0.000 title claims abstract description 51
- 108010001336 Horseradish Peroxidase Proteins 0.000 title claims abstract description 47
- 238000002360 preparation method Methods 0.000 title claims abstract description 15
- PTFCDOFLOPIGGS-UHFFFAOYSA-N Zinc dication Chemical compound [Zn+2] PTFCDOFLOPIGGS-UHFFFAOYSA-N 0.000 claims abstract description 12
- 238000011068 loading method Methods 0.000 claims abstract description 9
- 239000000243 solution Substances 0.000 claims description 63
- 238000003756 stirring Methods 0.000 claims description 28
- 239000011259 mixed solution Substances 0.000 claims description 21
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 17
- 239000008367 deionised water Substances 0.000 claims description 13
- 229910021641 deionized water Inorganic materials 0.000 claims description 13
- 241000255789 Bombyx mori Species 0.000 claims description 9
- 238000005406 washing Methods 0.000 claims description 7
- 238000001914 filtration Methods 0.000 claims description 6
- 239000012528 membrane Substances 0.000 claims description 6
- 239000011148 porous material Substances 0.000 claims description 6
- JIAARYAFYJHUJI-UHFFFAOYSA-L zinc dichloride Chemical compound [Cl-].[Cl-].[Zn+2] JIAARYAFYJHUJI-UHFFFAOYSA-L 0.000 claims description 6
- ONDPHDOFVYQSGI-UHFFFAOYSA-N zinc nitrate Chemical compound [Zn+2].[O-][N+]([O-])=O.[O-][N+]([O-])=O ONDPHDOFVYQSGI-UHFFFAOYSA-N 0.000 claims description 6
- 238000002156 mixing Methods 0.000 claims description 4
- 239000000126 substance Substances 0.000 claims description 4
- ZOIORXHNWRGPMV-UHFFFAOYSA-N acetic acid;zinc Chemical compound [Zn].CC(O)=O.CC(O)=O ZOIORXHNWRGPMV-UHFFFAOYSA-N 0.000 claims description 3
- 239000004246 zinc acetate Substances 0.000 claims description 3
- 239000011592 zinc chloride Substances 0.000 claims description 3
- 235000005074 zinc chloride Nutrition 0.000 claims description 3
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 claims description 3
- 229960001763 zinc sulfate Drugs 0.000 claims description 3
- 229910000368 zinc sulfate Inorganic materials 0.000 claims description 3
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 abstract description 20
- 238000000034 method Methods 0.000 abstract description 11
- 238000006243 chemical reaction Methods 0.000 abstract description 6
- 239000003937 drug carrier Substances 0.000 abstract description 6
- 239000003960 organic solvent Substances 0.000 abstract description 2
- 239000002245 particle Substances 0.000 description 9
- 102000004190 Enzymes Human genes 0.000 description 7
- 108090000790 Enzymes Proteins 0.000 description 7
- 229940088598 enzyme Drugs 0.000 description 7
- 239000004005 microsphere Substances 0.000 description 7
- 239000013078 crystal Substances 0.000 description 4
- 239000000706 filtrate Substances 0.000 description 4
- 229920005610 lignin Polymers 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 238000004132 cross linking Methods 0.000 description 3
- 239000000017 hydrogel Substances 0.000 description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 239000002131 composite material Substances 0.000 description 2
- 239000011258 core-shell material Substances 0.000 description 2
- 239000003431 cross linking reagent Substances 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000000839 emulsion Substances 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- 239000003642 reactive oxygen metabolite Substances 0.000 description 2
- 238000001878 scanning electron micrograph Methods 0.000 description 2
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- 229920001661 Chitosan Polymers 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 108010015776 Glucose oxidase Proteins 0.000 description 1
- 239000004366 Glucose oxidase Substances 0.000 description 1
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000010415 colloidal nanoparticle Substances 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- ZOMNIUBKTOKEHS-UHFFFAOYSA-L dimercury dichloride Chemical class Cl[Hg][Hg]Cl ZOMNIUBKTOKEHS-UHFFFAOYSA-L 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 230000001804 emulsifying effect Effects 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 229910021397 glassy carbon Inorganic materials 0.000 description 1
- 229940116332 glucose oxidase Drugs 0.000 description 1
- 235000019420 glucose oxidase Nutrition 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 239000003273 ketjen black Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000002088 nanocapsule Substances 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 230000020477 pH reduction Effects 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 229940043263 traditional drug Drugs 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/43—Enzymes; Proenzymes; Derivatives thereof
- A61K38/44—Oxidoreductases (1)
- A61K38/443—Oxidoreductases (1) acting on CH-OH groups as donors, e.g. glucose oxidase, lactate dehydrogenase (1.1)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/5005—Wall or coating material
- A61K9/5021—Organic macromolecular compounds
- A61K9/5052—Proteins, e.g. albumin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y111/00—Oxidoreductases acting on a peroxide as acceptor (1.11)
- C12Y111/01—Peroxidases (1.11.1)
- C12Y111/01007—Peroxidase (1.11.1.7), i.e. horseradish-peroxidase
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicinal Chemistry (AREA)
- Organic Chemistry (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Biochemistry (AREA)
- Epidemiology (AREA)
- General Engineering & Computer Science (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Toxicology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Gastroenterology & Hepatology (AREA)
- Immunology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Genetics & Genomics (AREA)
- Cosmetics (AREA)
Abstract
The invention discloses a silk fibroin microcapsule loading horseradish peroxidase and a preparation method thereof. The adopted method is based on that silk fibroin microcapsules are prepared in a zinc ion solution by a one-step method, the silk fibroin microcapsules are taken as a carrier, horseradish peroxidase solution is added in the process of forming the microcapsules, and the silk fibroin microcapsules loaded with the horseradish peroxidase are prepared by reaction under mild and zinc ion conditions. The invention has the advantages that: the preparation method has the advantages of simple process, mild reaction conditions, no need of organic solvent in the whole process, good biocompatibility and degradability of the prepared microcapsule, high loading rate of horseradish peroxidase, good degradability of hydrogen peroxide with a certain concentration, and hopeful application in the field of drug carriers.
Description
Technical Field
The invention relates to a silk fibroin microcapsule loading horseradish peroxidase and a preparation method thereof, belonging to the field of new materials.
Background
With the development of scientific technology, more and more studies have found that diseases have different microenvironments from normal tissues, such as Reactive Oxygen Species (ROS) level and hydrogen peroxide (H) in tumor tissues2O2) High concentration of hydrogen peroxide (H) significantly higher than normal tissue2O2) Can cause local active oxygen damage and even cell death. Therefore, it is becoming more and more important to find a drug carrier capable of reducing the active oxygen content of the diseased region. With the development of microcapsules, research on microcapsule technology has made great progress, not only many new patents on microcapsule synthesis technology are published, but also nanocapsules with particle size in the nanometer range are developed, and in recent years, microcapsules as drug carriers show good application prospects in the aspects of targeting of diseased cells and controllable release of drugs.
However, at present, the microcapsules capable of being used for loading drugs generally have the problems of complex preparation method and complex process. Chinese patent (CN 10931046 a) "a lignin wall material microcapsule based on Pickering emulsion interfacial crosslinking and its preparation and application in drug carriers" lignin particle dispersion is prepared by alkali dissolution and acidification of lignin in water, and water phase crosslinking agent is added as water phase; dissolving an oil phase cross-linking agent in an organic solvent to serve as an oil phase; mixing the water phase and the oil phase, and emulsifying to prepare O/W type Pickering emulsion; adding a catalyst, and heating to perform a crosslinking reaction to obtain a lignin wall material microcapsule; chinese patent (CN 102199592A) "a method for preparing co-immobilized glucose oxidase/catalase microspheres," using chitosan-arginine anion microspheres as a carrier, adding and mixing a mixed solution and glutaraldehyde, and preparing co-immobilized microspheres through cross-linking reaction, filtering, washing, and freeze-drying; chinese patent (CN 102380335A) "core-shell type hydrogel colloidal crystal microsphere and its preparation method and application" the colloidal crystal microsphere is used as template, filled with hydrogel, stripped, and then the colloidal nanoparticles in the microsphere are gradually corroded inwards to obtain the core-shell type hydrogel colloidal crystal microsphere, and its morphology and crystal structure are described. At present, no related process technology of a composite material which takes silk fibroin hollow particles as a raw material and is coated with horseradish peroxidase in the silk fibroin hollow particles is found.
The preparation method of the silk fibroin hollow microcapsule is simple, the source is rich, the biocompatibility is good, the horse radish peroxidase is a common enzyme in clinical test reagents, and the enzyme is not only widely used for a plurality of biochemical detection projects, but also widely used in immune (ELISA) kits. The preparation method of the microcapsule is simple and efficient, and the obtained silk fibroin microcapsule loading the horseradish peroxidase has good biocompatibility and degradability, has high horseradish peroxidase loading rate, has good degradation capability on hydrogen peroxide with a certain concentration, and is expected to be applied in the field of drug carriers.
Disclosure of Invention
The invention provides a horseradish peroxidase-loaded silk fibroin microcapsule and a preparation method thereof, aiming at overcoming the problems that the preparation process of the traditional drug carrier microcapsule is not environment-friendly, the method is complicated and the like, and simultaneously realizing the resource utilization of silk fibroin resources to prepare a new material for effectively reducing the concentration of hydrogen peroxide.
In order to achieve the purpose, the technical scheme of the invention is as follows:
1. a silk fibroin microcapsule loading horseradish peroxidase comprises a carrier and a coated substance, and is characterized in that: the carrier is a silk fibroin hollow microcapsule, and the coated substance is horseradish peroxidase.
2. A preparation method of a silk fibroin microcapsule loaded with horseradish peroxidase is suitable for the silk fibroin microcapsule loaded with the horseradish peroxidase, and is characterized by comprising the following steps:
1) preparing 0.2-10% silk fibroin solution by mass, stirring for 3-7 h at the rotating speed of 100-300 r/min, preparing 180-1500 mM zinc ion solution, and filtering the two solutions by microporous filter membranes with the pore diameter of 0.22 mu m respectively to obtain filtered silk fibroin solution and zinc ion solution;
2) mixing the filtered silk fibroin solution obtained in the step 1), zinc ion solution and deionized water according to a volume ratio of 4: 6: 10 preparing to obtain a mixed solution;
3) preparing a horseradish peroxidase solution with the mass concentration of 1mg/mL, and stirring for 1.5-2 h at 100 revolutions per minute under a dark condition to obtain a horseradish peroxidase solution;
4) adding 3mL of the horseradish peroxidase solution obtained in the step 3) into the mixed solution obtained in the step 2), uniformly stirring, adjusting the pH value of the mixed solution to 6.5-7.5, placing the mixed solution in a 37 ℃ thermostat, and stirring for 24 hours at the rotating speed of 100-300 r/min to obtain a reacted solution;
5) and (4) centrifugally washing the reacted solution obtained in the step 4) for 3 times by using deionized water to obtain the horseradish peroxidase-loaded silk fibroin microcapsule.
The molecular weight distribution of the silk fibroin is 5-30 KDa, and the silk fibroin can be silk fibroin from silkworms, tussah and other wild silks; the zinc ion solution can be any one of zinc nitrate, zinc chloride, zinc sulfate and zinc acetate solution.
Compared with the background art, the invention has the beneficial effects that:
the silk fibroin hollow microcapsule is prepared in a zinc ion solution by a one-step method, and horseradish peroxidase is added in the microcapsule forming process to realize the purpose of loading.
Drawings
FIG. 1 is a scanning electron micrograph (magnification: 2560) of a horseradish peroxidase-loaded field emission prepared in example 1.
FIG. 2 is a scanning electron micrograph (magnification: 20000 times) of a horseradish peroxidase-loaded field emission prepared in example 1.
FIG. 3 is a graph showing the redox current of the hydrogen peroxide solution after addition of the enzyme-supporting microcapsules prepared in example 1, measured electrochemically at various time points.
Detailed Description
The present invention will be further described with reference to the following specific examples.
Example 1:
1) preparing 80mL of a bombyx mori silk fibroin solution with the molecular weight distribution of 15KDa and the mass fraction of 0.2%, stirring for 4 hours at 200 revolutions per minute, preparing 120mL of a zinc nitrate solution with the concentration of 180mM, filtering the two solutions by using microporous filter membranes with the pore diameter of 0.22 mu m respectively, adding 200mL of deionized water into the filtrate, and uniformly stirring the mixed solution;
2) preparing a horseradish peroxidase solution with the mass concentration of 1mg/mL, and stirring for 2 hours at 100 revolutions per minute in the dark;
3) adding 3mL of the enzyme solution obtained in the step 2) into the mixed solution obtained in the step 1), uniformly stirring, adjusting the pH value of the new mixed solution to 7.0, and stirring for 24 hours in a 37 ℃ thermostat at the rotating speed of 180 r/min;
4) and (3) centrifugally washing the solution reacted in the step 3) for 3 times by using deionized water to obtain the horseradish peroxidase-loaded silk fibroin microcapsule.
The silk fibroin microcapsule loaded with horseradish peroxidase obtained in the example is silkworm cocoon type, the particle size is about 1 μm, and the uniformity is good.
Example 2:
1) 160mL of tussah silk fibroin solution with the molecular weight distribution of 5KDa and the mass fraction of 3% is prepared, the mixture is stirred for 5 hours at 300 r/min, 240mL of zinc chloride solution with the concentration of 1500mM is prepared, the two solutions are respectively filtered by microporous filter membranes with the pore diameter of 0.22 mu m, 400mL of deionized water is added into the filtrate, and the mixed solution is stirred uniformly;
2) preparing a horseradish peroxidase solution with the mass concentration of 1mg/mL, and stirring for 1.5h at 100 rpm in the dark;
3) adding 3mL of the enzyme solution obtained in the step 2) into the mixed solution obtained in the step 1), uniformly stirring, adjusting the pH value of the new mixed solution to 6.5, and stirring for 24 hours in a 37 ℃ thermostat at the rotating speed of 250 rpm;
4) and (3) centrifugally washing the solution reacted in the step 3) for 3 times by using deionized water to obtain the horseradish peroxidase-loaded silk fibroin microcapsule.
The silk fibroin microcapsules loaded with horseradish peroxidase obtained in the example are silkworm cocoon type and spherical, the particle size is different from 700nm to 1000nm, and the proportion of the silkworm cocoon type microcapsules is smaller than that of the silk fibroin microcapsules in the example 1.
Example 3:
1) preparing a tussore silk fibroin solution with the molecular weight distribution of 30KDa and the mass fraction of 10%, stirring for 7h at 100 r/min of 160mL, preparing 240mL of zinc sulfate solution with the concentration of 1000mM, filtering the two solutions by using microporous filter membranes with the pore diameter of 0.22 mu m respectively, adding 400mL of deionized water into the filtrate, and stirring the mixed solution uniformly;
2) preparing a horseradish peroxidase solution with the mass concentration of 1mg/mL, and stirring for 1.5h at 100 rpm in the dark;
3) adding 3mL of the enzyme solution obtained in the step 2) into the mixed solution obtained in the step 1), uniformly stirring, adjusting the pH value of the new mixed solution to 7.5, and stirring for 24 hours in a 37 ℃ thermostat at the rotating speed of 300 r/min;
4) and (3) centrifugally washing the solution reacted in the step 3) for 3 times by using deionized water to obtain the horseradish peroxidase-loaded silk fibroin microcapsule.
The silk fibroin microcapsule loaded with the horseradish peroxidase obtained in the embodiment is silkworm cocoon type and ball type, and the particle size is different from 700nm to 1000 nm.
Example 4:
1) preparing 160mL of a bombyx mori silk fibroin solution with the molecular weight distribution of 5KDa and the mass fraction of 5%, stirring for 3h at 100 r/min, preparing 240mL of a zinc acetate solution with the concentration of 1000mM, filtering the two solutions by using microporous filter membranes with the pore diameter of 0.22 mu m respectively, adding 400mL of deionized water into the filtrate, and uniformly stirring the mixed solution;
2) preparing a horseradish peroxidase solution with the mass concentration of 1mg/mL, and stirring for 2 hours at 100 revolutions per minute in the dark;
3) adding 3mL of the enzyme solution obtained in the step 2) into the mixed solution obtained in the step 1), uniformly stirring, adjusting the pH value of the new mixed solution to 7.5, and stirring for 24 hours in a 37 ℃ thermostat at the rotating speed of 100 rpm;
4) and (3) centrifugally washing the solution reacted in the step 3) for 3 times by using deionized water to obtain the horseradish peroxidase-loaded silk fibroin microcapsule.
The silk fibroin microcapsule loaded with the horseradish peroxidase obtained in the embodiment is mostly spherical, and the particle size is about 600 nm.
As the ratio of the zinc ion concentration to the silk fibroin concentration increases, the ratio of the silk fibroin microcapsules to the cocoon type increases, and the particle size gradually stabilizes at about 1000nm, as obtained in example 1, example 2, example 3, and example 4; the reaction solution is neutral, and the microcapsule becomes a silkworm cocoon type with good uniformity when the reaction stirring speed is 180 r/min.
The degrading ability of the horseradish peroxidase-loaded microcapsules was evaluated by measuring the concentration of hydrogen peroxide at different time periods after the enzyme-loaded microcapsules were added to hydrogen peroxide solutions of different concentrations.
1) Preparing an electrode: the working electrode is a glassy carbon electrode adsorbed with Ketjen black/chitosan/horseradish peroxidase composite material, the reference electrode is a saturated calomel electrode, and the counter electrode is a platinum mesh electrode;
2) electrochemically making hydrogen peroxide at 2 × 10 concentration-5～40×10-5Standard curve graph of concentration-current in mol/L;
3) the silk fibroin microcapsule loaded with horseradish peroxidase in example 1 was dissolved in deionized water to 3mL, and 1mL of the solution was added to 400mL of 5X 10-5mol/L (solution 1) and 40X 10-5Reacting in mol/L (solution 2) hydrogen peroxide solution under stirring;
4) and taking 60mL of measured solution current in the solution 1 and the solution 2 respectively at time points of 3h, 6 h, 9 h, 20 h and 24h after the microcapsules are added, and further obtaining the concentration of the hydrogen peroxide in the solution at different time points.
The redox currents in the solution measured at the various time points are shown in FIG. 3.
The foregoing lists merely illustrate specific embodiments of the invention. The present invention is not limited to the above embodiments, and many variations are possible. All modifications which can be derived or suggested by a person skilled in the art from the disclosure of the present invention are to be considered within the scope of the invention.
Claims (3)
1. A silk fibroin microcapsule loading horseradish peroxidase comprises a carrier and a coated substance, and is characterized in that: the carrier is a silk fibroin hollow microcapsule, and the coated substance is horseradish peroxidase.
2. A preparation method of a silk fibroin microcapsule loaded with horseradish peroxidase is suitable for the silk fibroin microcapsule loaded with the horseradish peroxidase, and is characterized by comprising the following steps:
1) preparing 0.2-10% silk fibroin solution by mass, stirring for 3-7 h at the rotating speed of 100-300 r/min, preparing 180-1500 mM zinc ion solution, and filtering the two solutions by microporous filter membranes with the pore diameter of 0.22 mu m respectively to obtain filtered silk fibroin solution and zinc ion solution;
2) mixing the filtered silk fibroin solution obtained in the step 1), zinc ion solution and deionized water according to a volume ratio of 4: 6: 10 preparing to obtain a mixed solution;
3) preparing a horseradish peroxidase solution with the mass concentration of 1mg/mL, and stirring for 1.5-2 h at 100 revolutions per minute under a dark condition to obtain a horseradish peroxidase solution;
4) adding 3mL of the horseradish peroxidase solution obtained in the step 3) into the mixed solution obtained in the step 2), uniformly stirring, adjusting the pH value of the mixed solution to 6.5-7.5, placing the mixed solution in a 37 ℃ thermostat, and stirring for 24 hours at the rotating speed of 100-300 r/min to obtain a reacted solution;
5) and (4) centrifugally washing the reacted solution obtained in the step 4) for 3 times by using deionized water to obtain the horseradish peroxidase-loaded silk fibroin microcapsule.
3. The preparation method of the horseradish peroxidase-loaded silk fibroin microcapsule according to claim 2, characterized in that: the molecular weight distribution of the silk fibroin is 5-30 KDa, and the silk fibroin can be silk fibroin from silkworms, tussah and other wild silks; the zinc ion solution can be any one of zinc nitrate, zinc chloride, zinc sulfate and zinc acetate solution.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108192731A (en) * | 2018-03-26 | 2018-06-22 | 上海应用技术大学 | A kind of orange flavor-fibroin albumen microcapsules and preparation method |
| CN108479650A (en) * | 2018-03-26 | 2018-09-04 | 上海应用技术大学 | A kind of osmanthus flower fragrance-fibroin albumen microcapsules and preparation method |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108192731A (en) * | 2018-03-26 | 2018-06-22 | 上海应用技术大学 | A kind of orange flavor-fibroin albumen microcapsules and preparation method |
| CN108479650A (en) * | 2018-03-26 | 2018-09-04 | 上海应用技术大学 | A kind of osmanthus flower fragrance-fibroin albumen microcapsules and preparation method |
Non-Patent Citations (1)
| Title |
|---|
| GUANGSHU WANG，等: "Reactive oxygen species-responsive silk sericin microcapsules used for antioxidative stress damage", 《MICROSC RES TECH.》 * |
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Application publication date: 20191220 |