CN106822038B - Preparation method and application of enzyme-coated silk nanospheres - Google Patents
Preparation method and application of enzyme-coated silk nanospheres Download PDFInfo
- Publication number
- CN106822038B CN106822038B CN201710046253.6A CN201710046253A CN106822038B CN 106822038 B CN106822038 B CN 106822038B CN 201710046253 A CN201710046253 A CN 201710046253A CN 106822038 B CN106822038 B CN 106822038B
- Authority
- CN
- China
- Prior art keywords
- silk
- enzyme
- solution
- nanospheres
- coated
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- 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
- A61K9/51—Nanocapsules; Nanoparticles
- A61K9/5107—Excipients; Inactive ingredients
- A61K9/5176—Compounds of unknown constitution, e.g. material from plants or animals
-
- 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)
-
- 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)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y101/00—Oxidoreductases acting on the CH-OH group of donors (1.1)
- C12Y101/03—Oxidoreductases acting on the CH-OH group of donors (1.1) with a oxygen as acceptor (1.1.3)
- C12Y101/03004—Glucose oxidase (1.1.3.4)
-
- 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)
- C12Y111/01006—Catalase (1.11.1.6)
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Wood Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Genetics & Genomics (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Immunology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Nanotechnology (AREA)
- Physics & Mathematics (AREA)
- Biomedical Technology (AREA)
- Botany (AREA)
- Optics & Photonics (AREA)
- Enzymes And Modification Thereof (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention discloses a preparation method of enzyme-coated silk nanospheres, which comprises the following steps: s1: preparing a silk solution; s2: wrapping enzyme into silk to form enzyme-wrapped silk nanospheres. The prepared silk nano-spheres can be applied to the medical field, in particular to the application of the silk nano-spheres in the anti-alcoholism drugs. The silk nano-spheres prepared by the method can effectively protect the activity of the encapsulated enzyme, realize the series reaction of a plurality of enzymes and have good biocompatibility.
Description
Technical Field
The invention relates to the technical field of enzyme coating, in particular to a preparation method and application of a silk nanosphere coated with enzyme.
Background
Enzymes are essential natural catalysts in organisms, and almost all biological reactions are closely related to the enzymes, such as protein synthesis and metabolism. However, natural biological enzymes are fragile, easily lose activity when exposed to normal temperature, and have high requirements on reaction environment, and it becomes very important to protect the activity of enzymes such as thermal stability and storage stability.
Moreover, most biological reactions are carried out by a series of reactions of various enzymes, and intermediate products are generated in the series of reactions, and most of the intermediate products are harmful to human bodies. Therefore, researchers can simulate in vivo biological tandem reaction by connecting or wrapping a plurality of enzymes together, so that not only can the reaction activity of the enzymes be effectively improved, but also toxic and harmful intermediate products, such as hydrogen peroxide generated by peroxidase catalytic reaction, can be effectively removed. However, most of the existing materials for wrapping enzymes are high molecular materials or novel inorganic materials, and the materials are poor in biocompatibility and not beneficial to use in animal bodies.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides a preparation method and application of silk nanospheres wrapping enzyme, so that the silk nanospheres which can effectively protect the activity of various enzymes, realize the series reaction among various enzymes and have good biocompatibility are prepared, and the silk nanospheres are applied to the medical field.
In order to achieve the purpose, the invention adopts the following technical scheme:
a preparation method of silk nano-spheres wrapping enzyme comprises the following steps:
s1, preparing a silk solution, which specifically comprises the following steps:
s11, cutting silkworm cocoon, and placing in 5% NaHCO solution3Boiling the solution, stirring with a glass rod for 30min, and repeating the steps twice;
s12, washing the silk obtained in the step S11 with deionized water at 60 ℃ for 20min, and repeating the step three times;
s13, drying the washed silk in an oven at 60 ℃, dissolving the dried silk in L iBr solution of 9.3M, and dissolving the silk for 4 hours at 60 ℃;
s14, dialyzing the silk solution obtained in the step S13 with deionized water for two days, and changing water every 2 hours;
s15, diluting the silk solution obtained in the step S14 to 6% w/w;
s2, wrapping enzyme into silk to form enzyme-wrapped silk nanospheres, which specifically comprises the following steps:
s21, mixing the silk solution prepared in the step S1 with an enzyme solution, and storing the mixture in a refrigerator at a low temperature for 4 to 8 hours, wherein the volume ratio of the silk solution to the enzyme solution is 5: 1;
s22, slowly dripping the mixed solution obtained in the step S21 into a certain amount of acetone in an amount of 25ul per drop, wherein the volume ratio of the mixed solution to the acetone is 1.2: 5, centrifuging the obtained solution at 18000rpm for 30 min;
s23, removing the supernatant, adding deionized water, and ultrasonically washing and dispersing;
s24, centrifuging the mixed solution 18000rpm obtained in the step S23 for 15 min;
s25, repeating the steps S23 and S24 three times to obtain the enzyme-coated silk nanospheres.
The prepared silk nanospheres comprise silk nanoshells and enzyme cores wrapped by the silk nanoshells, and the particle size of the silk nanospheres is 50-150 nm.
Preferably, the enzyme solution of step S21 includes one or more kinds of enzymes.
Preferably, the enzyme solution contains glucose oxidase and horseradish peroxidase in a mass ratio of 4: 1.
Preferably, the low temperature of step S21 is 4-10 ℃.
Preferably, the ultrasonic washing in step S23 is specifically: and (3) oscillating for 2min on an oscillator after carrying out 40% power ultrasound for 30s, and repeating for 3-5 times until the dispersion is uniform.
Preferably, the silk nanospheres can be applied in the medical field.
Preferably, the silk nanospheres can be used for preparing an anti-alcoholism drug, and the prepared anti-alcoholism drug is simultaneously coated with alcohol hydrogenase and catalase.
After adopting the technical scheme, compared with the background technology, the invention has the following advantages:
1. effectively protecting the activity of the encapsulated enzyme. Firstly, when the silk nanospheres are prepared, the silk solution and the enzyme solution are mixed and then stored in a low-temperature refrigerator at 4-8 ℃, so that the stability of the enzyme activity can be ensured, the enzyme is prevented from losing the activity in the preparation process, and the effective output rate of the silk nanospheres coated with the enzyme is ensured; secondly, the silk nanospheres are of a high-crystallinity structure, silk fibroin is contained in a silk shell for wrapping the enzyme, the silk fibroin has controllable degradability and is not easily degraded by protease in a human body, the activity of the enzyme can be protected for a long time, and a natural barrier is provided for the wrapped enzyme, so that the thermal stability and the storage stability of the wrapped enzyme are effectively protected; thirdly, the particle size of the prepared silk nanospheres is 50-150nm, so that the particle size can effectively improve the dispersibility of the silk nanospheres in an aqueous solution, and the high dispersibility ensures the high-efficiency catalytic reaction of enzyme; therefore, the activity of the encapsulated enzyme is strictly protected from the processes of preparation, storage and use.
2. Realize the tandem reaction of a plurality of enzymes. In the tandem reaction, since the intermediate product is transported among a plurality of enzymes, and the reaction environment is usually a high-viscosity solution, such as cell sap, blood, etc., the high-viscosity solution is more unfavorable for the transportation of the intermediate product, and the tandem reaction is hindered. The silk nanospheres can wrap various related enzymes at the same time, the distance between the related enzymes can be greatly reduced through wrapping, the influence of solution viscosity on the transmission of reaction intermediate products is reduced, the activity of the enzymes is better exerted, toxic and harmful intermediate products are effectively removed, in addition, the activity of the related enzymes can be further enhanced by adjusting the mass ratio of different wrapped enzymes, and the superiority of the silk nanospheres in series reaction is realized.
3. Has good biocompatibility. The silk fibroin contained in the silk nanospheres prepared by the invention is a biological material which is certified by FDA, has good biocompatibility, cannot cause any rejection reaction in a human body, cannot cause inflammation, avoids the defects of difficult decomposition, rejection and the like, and can be applied to the medical field to act on animal bodies.
Drawings
FIG. 1 is an electron microscope image of the silk nanospheres of the present invention
FIG. 2 measurement results of thermostability and storage stability of Encapsulated enzyme
FIG. 3 tandem reaction results of encapsidated enzymes
FIG. 4 comparative results of relative enzyme activities
FIG. 5 Activity assay of Encapsulated enzyme in high viscosity solution
FIG. 6 determination of antialcoholism effect of enzyme-coated silk nanospheres
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Example 1 preparation of enzyme-Encapsulated Silk nanospheres
A preparation method of silk nano-spheres wrapping enzyme comprises the following steps:
s1, preparing a silk solution, which specifically comprises the following steps:
s11, cutting silkworm cocoon, and placing in 5% NaHCO solution3Boiling the solution, stirring with a glass rod for 30min, and repeating the steps twice;
s12, washing the silk obtained in the step S11 with deionized water at 60 ℃ for 20min, and repeating the step three times;
s13, drying the washed silk in an oven at 60 ℃, dissolving the dried silk in L iBr solution of 9.3M, and dissolving the silk for 4 hours at 60 ℃;
s14, dialyzing the silk solution obtained in the step S13 with deionized water for two days, and changing water every 2 hours;
s15, diluting the silk solution obtained in the step S14 to 6% w/w;
s2, wrapping enzyme into silk to form enzyme-wrapped silk nanospheres, which specifically comprises the following steps:
s21, mixing 1ml of the silk solution prepared in the step S1 with 200ul of enzyme solution containing alcohol enzyme and catalase, and placing the mixture in a refrigerator at 4 ℃ for low-temperature storage for 6 hours;
s22, slowly dripping 1ml of the mixed solution obtained in the step S21 into 5ml of acetone in an amount of 25ul per drop, and centrifuging the obtained mixed solution at 18000rpm for 30 min;
s23, removing the supernatant, adding deionized water, performing 40% power ultrasound for 30S, then oscillating on an oscillator for 2min, repeating for 3-5 times until the dispersion is uniform;
s24, centrifuging the mixed solution 18000rpm obtained in the step S23 for 15 min;
s25, repeating the steps S23 and S24 three times to obtain the enzyme-coated silk nanospheres.
The particle size of the silk nanospheres is 50-150 nm.
The electron microscope image of the prepared enzyme-coated silk nanospheres is shown in FIG. 1. The scale of fig. 1a is 500um and the scale of fig. 1b is 100 um.
Example 2 Activity assay of enzyme Encapsulated in Silk Nanobules
(1) Thermal and storage stability of the enzyme
The thermal stability and storage stability of the enzymes before and after coating were compared by determining the relative enzyme activity of the enzyme without silk coating and the enzyme with silk coating at different time points. The results are shown in FIG. 2.
FIG. 2a shows the relative enzyme activities of glucose oxidase without and with silk encapsulation at 60 ℃. As can be seen from the figure, at 60 ℃ the glucose oxidase without silk coating lost half the activity for 1 hour; the glucose oxidase wrapped by the silk loses half of activity after 30 hours; therefore, the thermal stability of the enzyme coated by the silk nanospheres is effectively protected.
FIG. 2b shows the storage stability of horseradish peroxidase without and with silk coating at 25 ℃ on different days. As can be seen from the figure, at 25 ℃ the horse radish peroxidase without silk coating lost half of the activity after 4 days and completely lost the activity after 14 days; the activity of the horseradish peroxidase wrapped by the silk is about 70 percent after 14 days; therefore, the storage stability of the enzyme coated by the silk nanospheres is effectively protected.
(2) Superiority of coated enzyme in tandem reaction
The simultaneous encapsulation of multiple enzymes into silk nanospheres will result in a tandem reaction, taking glucose oxidase as an example to catalyze the decomposition of glucose, and the results are shown in fig. 3.
As can be seen from fig. 3a, when only glucose oxidase (GOx) is added in the glucose decomposition reaction, a large amount of intermediate hydrogen peroxide is generated as the reaction proceeds, so that the concentration of hydrogen peroxide in the solution gradually increases; when glucose oxidase (GOx) and catalase (Cat) are added simultaneously, hydrogen peroxide, an intermediate product, is still generated as the glucose decomposition reaction proceeds, but the rate of increase in the hydrogen peroxide concentration is slightly decreased because catalase catalyzes the decomposition of hydrogen peroxide to reduce the amount of harmful hydrogen peroxide.
As can be seen from fig. 3b, in the glucose decomposition reaction, when only glucose oxidase coated with silk nanospheres is added, a large amount of hydrogen peroxide intermediate product is still generated, and the concentration of hydrogen peroxide in the solution gradually increases; when the silk nanospheres wrapped with glucose oxidase and catalase at the same time are added, the concentration of hydrogen peroxide in the solution is increased to a small extent along with the reaction, the increase speed is obviously slowed down, and the effect of removing harmful intermediate product hydrogen peroxide is obviously better than that of an experimental group without silk nanosphere wrapping (a result shown in figure 3 a).
This is because, in the tandem reaction of a plurality of enzymes, the transfer of reaction intermediates between the plurality of enzymes is involved, and the plurality of enzymes are packed together, so that the distance between them can be greatly reduced, the influence of the solution viscosity on the transfer of the reaction intermediates can be reduced, and higher activity can be preserved.
Further, when the mass ratio of glucose oxidase and horseradish peroxidase was different when they were mixed, the relative enzyme activity was measured, and the results are shown in fig. 4. Experiments prove that: when the mass ratio of the glucose oxidase to the horseradish peroxidase wrapped in the silk nanospheres is different, the relative enzyme activity is also different, and when the mass ratio of the glucose oxidase to the horseradish peroxidase is 4:1, the activity is highest. Therefore, the activity of the finally obtained silk nanospheres coated with the enzyme can be optimized by controlling the mass ratio of the enzymes.
Therefore, the enzyme coated by the silk nanospheres can effectively remove toxic and harmful intermediate products such as hydrogen peroxide in the series reaction process, the effect is obviously superior to that of the enzyme without the silk nanospheres, and the superiority of the coated enzyme in the series reaction is reflected.
(3) Encapsulated enzyme activity in high viscosity solutions
The enzyme without the silk nanosphere coating and the enzyme with the silk nanosphere coating were placed in polyethylene glycol (PEG) solutions of different concentrations, and the relative enzyme activities of the enzymes at high viscosity were tested, and the results are shown in fig. 5. It can be seen from the figure that the solution viscosity gradually increases with the increase of PEG concentration, and the relative activity of the enzyme gradually decreases, but when the PEG concentration is higher than 15 v/v%, that is, in the high-viscosity solution, the relative activity of the enzyme coated with the silk nanospheres decreases at a speed obviously lower than that of the enzyme without the silk nanospheres, which indicates that the coating of the silk nanospheres can protect the activity of the enzyme in the high-viscosity solution (such as cytosol, blood, etc.), and the enzyme can be stored with higher activity under the same conditions.
Example 3 application in anti-hangover medicine
The alcohol oxidase (AOx) and catalase (Cat) are wrapped in the silk nano-spheres to prepare the protease-resistant anti-alcoholism drug, and the effect of the protease-resistant anti-alcoholism drug is shown in figure 6.
Adding alcoholic enzyme without silk nano-spheres coating or alcoholic enzyme with silk nano-spheres coating into the intestinal juice simulated solution, comparing relative enzyme activities of the two solutions, as shown in FIG. 6a, the alcoholic enzyme with silk nano-spheres coating still has more than 90% of activity after 2h, and the relative activity of the enzyme without silk nano-spheres coating is rapidly reduced to 5% after 20 min. The medicine containing protease reaches intestinal juice after being taken orally, the protease in the intestinal juice can promote the hydrolysis of protein, most of the enzyme is protein, so that the enzyme is rapidly decomposed and inactivated in the intestinal juice, and the silk nano-spheres coated outside the alcoholic enzyme can effectively protect the alcoholic enzyme from being decomposed, so that the activity of the enzyme is protected. Therefore, the silk nanospheres can effectively prevent the protease from hydrolyzing the wrapped enzyme, and the exertion of the enzyme activity is ensured.
And simultaneously adding alcoholic enzyme and catalase without the coating of the silk nano-spheres or adding the silk nano-spheres coated with the alcoholic enzyme and the catalase into the intestinal juice simulation solution, and comparing the change of the alcohol concentration with the lapse of time. As shown in fig. 6b, the alcoholic enzyme and catalase without silk coating could not explain alcohol, and the alcohol concentration still did not change significantly after 6 h; and the alcohol enzyme and the catalase coated with the silk nano-spheres can effectively degrade alcohol, so that the alcohol concentration is rapidly reduced, the alcohol concentration is reduced to 3% after 6 hours, and the anti-alcohol effect is remarkable.
Therefore, the silk nano-spheres wrapped with the alcoholic enzyme and the catalase can effectively protect the wrapped enzyme from being decomposed by the protease, can efficiently reduce the alcohol concentration in a human body, and achieves a remarkable anti-alcohol effect.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are included in the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.
Claims (6)
1. A preparation method of silk nano-spheres wrapping enzyme is characterized by comprising the following steps: the method comprises the following steps:
s1, preparing a silk solution, which specifically comprises the following steps:
s11, cutting silkworm cocoon, and placing in 5% NaHCO solution3Boiling the solution, stirring with a glass rod for 30min, and repeating the steps twice;
s12, washing the silk obtained in the step S11 with deionized water at 60 ℃ for 20min, and repeating the step three times;
s13, drying the washed silk in an oven at 60 ℃, dissolving the dried silk in L iBr solution of 9.3M, and dissolving the silk for 4 hours at 60 ℃;
s14, dialyzing the silk solution obtained in the step S13 with deionized water for two days, and changing water every 2 hours;
s15, diluting the silk solution obtained in the step S14 to 6% w/w;
s2, wrapping enzyme into the silk to form enzyme-wrapped silk nanospheres;
s21, mixing the silk solution prepared in the step S1 with an enzyme solution, and storing the mixture in a refrigerator at a low temperature for 4 to 8 hours, wherein the volume ratio of the silk solution to the enzyme solution is 5: 1; the enzyme solution contains glucose oxidase and horse radish peroxidase;
s22, slowly dripping the mixed solution obtained in the step S21 into a certain amount of acetone in an amount of 25ul per drop, wherein the volume ratio of the mixed solution to the acetone is 1.2: 5, centrifuging the obtained solution at 18000rpm for 30 min;
s23, removing the supernatant, adding deionized water, and ultrasonically washing and dispersing;
s24, centrifuging the mixed solution 18000rpm obtained in the step S23 for 15 min;
s25, repeating the steps S23 and S24 for three times to obtain the enzyme-coated silk nanospheres;
the prepared silk nanospheres comprise silk nanoshells and enzyme cores wrapped by the silk nanoshells, and the particle size of the silk nanospheres is 50-150 nm.
2. The method for preparing enzyme-coated silk nanospheres according to claim 1, wherein the enzyme-coated silk nanospheres are prepared by the following steps: the mass ratio of the glucose oxidase to the horseradish peroxidase is 4: 1.
3. The method for preparing enzyme-coated silk nanospheres according to claim 1, wherein the enzyme-coated silk nanospheres are prepared by the following steps: the low temperature in step S21 is 4-10 ℃.
4. The method for preparing enzyme-coated silk nanospheres according to claim 1, wherein the enzyme-coated silk nanospheres are prepared by the following steps: the ultrasonic washing in step S23 specifically includes: and (3) oscillating for 2min on an oscillator after carrying out 40% power ultrasound for 30s, and repeating for 3-5 times until the dispersion is uniform.
5. Use of enzyme coated silk nanospheres according to any of claims 1 to 4 wherein: the silk nanospheres are applied to preparation of medicines.
6. The use of the enzyme-coated silk nanospheres according to claim 5, wherein: the application of the silk nano-spheres in preparing the anti-alcoholism drug is characterized in that the prepared anti-alcoholism drug is simultaneously coated with alcohol hydrogenase and catalase.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710046253.6A CN106822038B (en) | 2017-01-22 | 2017-01-22 | Preparation method and application of enzyme-coated silk nanospheres |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710046253.6A CN106822038B (en) | 2017-01-22 | 2017-01-22 | Preparation method and application of enzyme-coated silk nanospheres |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106822038A CN106822038A (en) | 2017-06-13 |
| CN106822038B true CN106822038B (en) | 2020-08-07 |
Family
ID=59119415
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710046253.6A Active CN106822038B (en) | 2017-01-22 | 2017-01-22 | Preparation method and application of enzyme-coated silk nanospheres |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106822038B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1560115A (en) * | 2004-03-10 | 2005-01-05 | 复旦大学 | Nano microball of shombycin protein and preparation process thereof |
| CN1560136A (en) * | 2004-03-04 | 2005-01-05 | 苏州大学 | Manufacture process of nano fibroin partical |
| CN1834240A (en) * | 2006-03-30 | 2006-09-20 | 苏州大学 | Silk nano granular of immobilized enzyme, and prepn. process thereof |
-
2017
- 2017-01-22 CN CN201710046253.6A patent/CN106822038B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1560136A (en) * | 2004-03-04 | 2005-01-05 | 苏州大学 | Manufacture process of nano fibroin partical |
| CN1560115A (en) * | 2004-03-10 | 2005-01-05 | 复旦大学 | Nano microball of shombycin protein and preparation process thereof |
| CN1834240A (en) * | 2006-03-30 | 2006-09-20 | 苏州大学 | Silk nano granular of immobilized enzyme, and prepn. process thereof |
Non-Patent Citations (4)
| Title |
|---|
| Bioconjugation of Silk Fibroin Nanoparticles with Enzyme and Peptide and Their Characterization;Fei Wang,et al;《Advances in Protein Chemistry and Structural Biology》;20151231;第98卷;第264-286页 * |
| Stabilization of Enzymes in Silk Films;Shenzhou Lu,et al;《Biomacromolecules》;20091231;第10卷(第5期);第1032-1042页 * |
| 丝素作为固定化酶载体的研究进展与应用;陈艳芳等;《广东蚕业》;20080331;第38卷(第1期);第44-49页 * |
| 家蚕丝素固定化过氧化氢酶的制备及其理化特性的研究;朱祥瑞等;《农业生物技术学报》;19991231;第7卷(第3期);第277-280页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106822038A (en) | 2017-06-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| BE1014638A6 (en) | Method of preparation of derivatives of cell wall from biomass. | |
| EP0270643B1 (en) | Method for conditioning an antibacterial composition and antibacterial composition thus conditioned | |
| Abd Manan et al. | Insight into the Rhizomucor miehei lipase supported on chitosan-chitin nanowhiskers assisted esterification of eugenol to eugenyl benzoate | |
| Lehmann et al. | Silk fibroin-based therapeutics for impaired wound healing | |
| EP2832352B1 (en) | Capsule disintegrable in large-intestine-specific manner | |
| Manrich et al. | Immobilization and stabilization of xylanase by multipoint covalent attachment on agarose and on chitosan supports | |
| EP2292752B9 (en) | Seamless capsule | |
| JP2015113331A (en) | Hydrogen-containing composition and hydrogen addition method using the hydrogen-containing composition | |
| CN109251330B (en) | Nano chitin-PVA hydrogel and preparation method and application thereof | |
| Elnashar et al. | Novel epoxy activated hydrogels for solving lactose intolerance | |
| Wu et al. | Coimmobilization of naringinases on silk fibroin nanoparticles and its application in food packaging | |
| CN109576256A (en) | Method for encapsulating double enzymes by magnetic DNA hydrogel | |
| Simó et al. | Highly efficient malolactic fermentation of red wine using encapsulated bacteria in a robust biocomposite of silica-alginate | |
| Sharmeen et al. | Application of polysaccharides in enzyme immobilization | |
| CN106822038B (en) | Preparation method and application of enzyme-coated silk nanospheres | |
| Sakai et al. | Both ionically and enzymatically crosslinkable alginate–tyramine conjugate as materials for cell encapsulation | |
| CN109999008B (en) | Composite agarose microspheres and preparation method and application thereof | |
| Maroufi et al. | Recent advances of macromolecular hydrogels for enzyme immobilization in the food products | |
| Du et al. | pH and amylase dual stimuli-responsive electrospinning membrane for smart releasing thymol to control melon Rhizopus stolonifer | |
| CN109913440A (en) | A method of passing through pressure synthesising biological enzyme/MOFs composite functional material | |
| Tincu et al. | An accessible method to improve the stability and reusability of porcine pancreatic α-amylase via immobilization in gellan-based hydrogel particles obtained by ionic cross-linking with Mg2+ ions | |
| CN107118286B (en) | The preparation method of chitosan crosslinked 2,5-furandaldehyde hydrogel | |
| CN114304143A (en) | Pesticide delivery system and preparation method thereof | |
| CN114931588B (en) | Preparation method of silver nanoparticle kanamycin antibacterial hydrogel | |
| CN104745662A (en) | Method for preparing hyaluronic acid substitute through oxidizing chitosan by laccase-TEMPO system |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |