CN106822038A - A kind of preparation method and applications of the silk nanometer bead for wrapping up enzyme - Google Patents
A kind of preparation method and applications of the silk nanometer bead for wrapping up enzyme Download PDFInfo
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- CN106822038A CN106822038A CN201710046253.6A CN201710046253A CN106822038A CN 106822038 A CN106822038 A CN 106822038A CN 201710046253 A CN201710046253 A CN 201710046253A CN 106822038 A CN106822038 A CN 106822038A
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- 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
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- 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)
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- 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)
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- 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)
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- 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)
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- 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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Abstract
The invention discloses a kind of preparation method of the silk nanometer bead for wrapping up enzyme, comprise the following steps：S1：Prepare silk solution；S2：Enzyme is wrapped up into silk, the silk nanometer bead of parcel enzyme is formed.Obtained silk nanometer bead can be applied to medical domain, the especially application on antialcoholic drug.Silk nanometer bead can effectively protect the activity of parcel enzyme according to obtained in the method for the invention, the cascade reaction of various enzymes be realized, while having good bio-compatibility.
Description
Technical field
The present invention relates to enzyme technique for packing field, and in particular to a kind of preparation method of the silk of parcel enzyme nanometer bead and
Its application.
Background technology
Enzyme is natural catalyst essential in organism, and almost all of biological respinse is all closely bound up with it, such as
Protein synthesis and metabolism etc..But natural biology enzyme is very fragile, easily loss of activity is exposed at normal temperatures, and to anti-
Answer environmental requirement very high, activity such as heat endurance and the storage stability of protective enzyme become particularly significant.
In addition most biological respinse is completed by the cascade reaction of various enzymes, in these tandem reactor process
In can produce intermediate product, and some intermediate products are mostly harmful.Therefore, researchers connect by by various enzymes
The method for connecing or being bundled together, the biological cascade reaction come in analogue body, the reaction that so can not only effectively improve enzyme is lived
Property, can also efficiently dispel poisonous and hazardous intermediate product, the hydrogen peroxide that such as Catalyzed Synthesis By Peroxidase reaction is produced.So
And, the material of existing parcel enzyme is generally macromolecular material or new inorganic material, and bio-compatibility is poor, is unfavorable for dynamic
Used in object.
The content of the invention
It is an object of the invention to overcome above-mentioned the deficiencies in the prior art, there is provided a kind of silk nanometer bead of parcel enzyme
Preparation method and applications, be obtained it is a kind of can effectively protect the activity of various enzymes simultaneously, realize that the series connection between various enzymes is anti-
Should, while but also with the silk nanometer bead of good bio-compatibility, and it is applied to medical domain.
To achieve the above object, the present invention uses following technical scheme：
A kind of preparation method of the silk nanometer bead for wrapping up enzyme, comprises the following steps：
S1, silk solution is prepared, specially：
S11, silk cocoon is shredded, be placed in 5% NaHCO3Boiled in solution, and 30min is stirred with glass bar, repeat the step
Suddenly twice；
S12, with gained silk 20min in 60 DEG C of deionized water washing step S11, repeat the step 3 time；
S13, washed silk is placed in 60 DEG C of baking ovens dry, be subsequently solubolized in the LiBr solution of 9.3M, 60 DEG C molten
Solution 4h；
Gained silk solution two days, changes a water in every 2 hours in S14, the step S13 that dialysed with deionized water；
S15, by step S14 gained silk solution be diluted to 6%w/w；
S2, enzyme is wrapped up into silk, form the silk nanometer bead of parcel enzyme, specially：
S21, silk solution obtained in step S1 is taken, be placed in after mixing with enzyme solutions Cord blood 4-8 hours in refrigerator, its
Middle silk solution is 5 with the volume ratio of enzyme solutions：1；
S22, gained mixed solution in step S21 is taken, be slowly dropped into a certain amount of acetone with the amount of every drop 25ul, wherein
Mixed solution is 1.2 with the volume ratio of acetone：5, resulting solution 18000rpm is centrifuged 30min；
S23, supernatant is abandoned, add deionized water, supersound washing simultaneously disperses；
S24, by gained mixed solution 18000rpm centrifugations 15min in step S23；
S25, repeat step S23, S24 tri- times, that is, be obtained the silk nanometer bead of parcel enzyme.
Obtained silk nanometer bead is made up of the enzyme core of silk nanometer shell and its parcel, and the silk nanometer is small
The particle diameter of ball is 50-150nm.
Preferably, the enzyme class for being included in the enzyme solutions described in step S21 is one or more.
Preferably, the mass ratio containing glucose oxidase and horseradish peroxidase in the enzyme solutions is 4:1.
Preferably, the low temperature described in step S21 is 4-10 DEG C.
Preferably, supersound washing described in step S23 is specially：2min is shaken on the oscillator after 40% power ultrasonic 30s,
Repeat 3-5 times, until being uniformly dispersed.
Preferably, the silk nanometer bead can be in medical domain application.
Preferably, the silk nanometer bead can be used to prepare antialcoholic drug, wrap up alcohol in obtained antialcoholic drug simultaneously
Hydrogenase and catalase.
After adopting the above technical scheme, the present invention is compared with background technology, have the following advantages that：
1st, effectively the activity of enzyme is wrapped up in protection.First, it is of the invention when silk nanometer bead is prepared, by silk solution and enzyme
4-8 DEG C of low temperature refrigerator is placed in after solution mixing to preserve, it can be ensured that the stabilization of enzymatic activity, it is to avoid enzyme just loses in preparation process
Deactivation, so as to ensure that the throughput rate of the silk nanometer bead of parcel enzyme；Secondly, silk nanometer of the present invention is small
Ball is the structure of high-crystallinity, and it is used to contain fibroin albumen in the silk shell for wrap up enzyme, and fibroin albumen has controlled degradation
Property, be difficult to be easily degraded by proteases in human body, can protective enzyme for a long time activity, for the enzyme for being wrapped up provides natural screen
Barrier, so as to effectively protect the heat endurance and storage stability of wrapped up enzyme；Again, due to obtained silk nanometer bead
Particle diameter in 50-150nm, the particle size can effectively improve silk nanometer bead dispersiveness in aqueous, and high score
Scattered property ensure that the efficient catalytic reaction of enzyme；Therefore, the present invention from be prepared into preserve again to the process for using all strict protections
Wrap up the activity of enzyme.
2nd, the cascade reaction of various enzymes is realized.In cascade reaction, because intermediate product can be transmitted between various enzymes, and
Reaction environment is usually high viscosity solution, such as cell liquid, blood, and such high viscosity solution is less useful for the biography of intermediate product
It is defeated, hinder the carrying out of cascade reaction.Silk nanometer bead of the present invention can simultaneously wrap up various relevant enzymes, by parcel
The distance between relevant enzyme can be substantially reduced, the influence that solution viscosity is transmitted to reaction intermediate be reduced, so as to ensure enzyme
Activity preferably play, effectively remove poisonous and hazardous intermediate product, additionally, regulation it is different parcel enzymes between mass ratioes can
The activity of relevant enzyme is further enhanced, superiority of the silk nanometer bead in cascade reaction is realized.
3rd, with good bio-compatibility.The fibroin albumen contained in silk nanometer bead obtained by the present invention is one
Plant by the biomaterial of FDA certifications, with good bio-compatibility, any rejection will not be caused in human body,
Inflammation will not be caused, it is to avoid be difficult to the shortcomings of decomposing and repel, such that it is able to acting on animal body in medical domain application.
Brief description of the drawings
The electron microscope of Fig. 1 silk nanometer beads of the present invention
Fig. 2 wraps up the heat endurance and storage stability measurement result of enzyme
Fig. 3 wraps up the cascade reaction result of enzyme
Fig. 4 relative activity comparative results
Fig. 5 wraps up determination of activity of the enzyme in high viscosity solution
The dispelling effects of alcohol of the silk nanometer bead of Fig. 6 parcel enzymes is determined
Specific embodiment
In order to make the purpose , technical scheme and advantage of the present invention be clearer, it is right below in conjunction with drawings and Examples
The present invention is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, and
It is not used in the restriction present invention.
Embodiment 1 prepares the silk nanometer bead of parcel enzyme
A kind of preparation method of the silk nanometer bead for wrapping up enzyme, comprises the following steps：
S1, silk solution is prepared, specially：
S11, silk cocoon is shredded, be placed in 5% NaHCO3Boiled in solution, and 30min is stirred with glass bar, repeat the step
Suddenly twice；
S12, with gained silk 20min in 60 DEG C of deionized water washing step S11, repeat the step 3 time；
S13, washed silk is placed in 60 DEG C of baking ovens dry, be subsequently solubolized in the LiBr solution of 9.3M, 60 DEG C molten
Solution 4h；
Gained silk solution two days, changes a water in every 2 hours in S14, the step S13 that dialysed with deionized water；
S15, by step S14 gained silk solution be diluted to 6%w/w；
S2, enzyme is wrapped up into silk, form the silk nanometer bead of parcel enzyme, specially：
S21, silk solution obtained in 1ml steps S1 is taken, the enzyme solutions for containing enzyme alcohol and catalase with 200ul are mixed
Cord blood 6 hours in 4 DEG C of refrigerators are placed in after conjunction；
S22, gained mixed solution in 1ml steps S21 is taken, be slowly dropped into 5ml acetone with the amount of every drop 25ul, by gained
Mixed solution 18000rpm is centrifuged 30min；
S23, supernatant is abandoned, add deionized water, shake 2min after 40% power ultrasonic 30s on the oscillator, repeat 3-5
It is secondary, until being uniformly dispersed；
S24, by gained mixed solution 18000rpm centrifugations 15min in step S23；
S25, repeat step S23, S24 tri- times, that is, be obtained the silk nanometer bead of parcel enzyme.
The particle diameter of obtained silk nanometer bead is 50-150nm.
The electron microscope of the silk nanometer bead of obtained parcel enzyme is as shown in Figure 1.The scale of Fig. 1 a is 500um, Fig. 1 b's
Scale is 100um.
The determination of activity of the enzyme of the silk of embodiment 2 nanometer bead parcel
(1) heat endurance and storage stability of enzyme
By determining the enzyme wrapped up without silk and the enzyme for thering is silk to wrap up in the relative activity of different time points, compare bag
Wrap up in the heat endurance and storage stability of front and rear enzyme.Result is as shown in Figure 2.
Fig. 2 a are represented at 60 DEG C, without the silk glucose oxidase for wrapping up and the glucose oxidase for having silk to wrap up
Relative activity.It can be seen that at 60 DEG C, the glucose oxidase without silk parcel just loses half in 1 hour
Activity；And there is the glucose oxidase that silk is wrapped up just to lose a semiactive after 30 hours；As can be seen here, it is small through silk nanometer
The heat endurance of the enzyme of ball parcel is effectively protected.
Fig. 2 b are represented at 25 DEG C, without the silk horseradish peroxidase for wrapping up and the horseradish peroxidase for having silk to wrap up
Storage stability of the enzyme in different number of days.It can be seen that at 25 DEG C, the horseradish peroxidase 4 without silk parcel
A semiactive is lost after it, activity completely loses after 14 days；And there is the horseradish peroxidase that silk is wrapped up also to retain after 14 days
70% or so activity；As can be seen here, the storage stability of the enzyme for being wrapped up through silk nanometer bead has obtained effective protection.
(2) superiority of the parcel enzyme in cascade reaction
Various enzymes are wrapped up into silk nanometer bead simultaneously, cascade reaction can be formed, grape is catalyzed with glucose oxidase
As a example by sugar decomposition, as a result as shown in Figure 3.
From Fig. 3 a as can be seen that in breakdown of glucose reaction, when only adding glucose oxidase (GOx), with anti-
The carrying out answered, can produce substantial amounts of intermediate product hydrogen peroxide so that the concentration of hydrogen peroxide gradually rises in solution；When simultaneously
When adding glucose oxidase (GOx) and catalase (Cat), with the carrying out that breakdown of glucose is reacted, still can produce
Intermediate product hydrogen peroxide, but because catalase energy catalyzing hydrogen peroxide is decomposed, so as to reduce harmful substance hydrogen peroxide
Amount, therefore the growth rate of concentration of hydrogen peroxide is slightly decreased.
From Fig. 3 b as can be seen that in breakdown of glucose reaction, the glucose wrapped up through silk nanometer bead is only added
During oxidizing ferment, substantial amounts of hydrogen peroxide intermediate product still can be produced, the concentration of hydrogen peroxide gradually rises in solution；Work as addition
The silk nanometer bead of glucose oxidase and catalase, as reaction is carried out, hydrogen peroxide in solution have been wrapped up simultaneously
Concentration increasing degree very little, growth rate substantially slows down, and its effect is significant for dispelling harmful intermediate product hydrogen peroxide is better than
Experimental group (result shown in Fig. 3 a) without silk nanometer bead parcel.
Because, in the cascade reaction of various enzymes, transmission of the reaction intermediates between various enzymes can be related to, and
Various enzymes are bundled together, the distance between they can be substantially reduced, when reduction solution viscosity is transmitted to reaction intermediates
Influence, so as to preserve activity higher.
Further, the mass ratio when glucose oxidase mixes from horseradish peroxidase is different, measures its relative
Enzymatic activity, as a result as shown in Figure 4.Experiment is proved：When the glucose oxidase and horseradish peroxidating that are wrapped up in silk nanometer bead
The mass ratio of thing enzyme is different, and its relative activity is also different, when the mass ratio of glucose oxidase and horseradish peroxidase is
4:When 1, active highest.It can be seen that, by controlling the mass ratio between various enzymes, the silk of the parcel enzyme for finally giving can be optimized
The activity of nanometer bead.
As can be seen here, the enzyme for being wrapped up by silk nanometer bead can be removed effectively in tandem reactor process, more poisonous to be had
Harmful intermediate product, such as hydrogen peroxide, its effect are substantially better than the enzyme without silk nanometer bead parcel, embody parcel enzyme and exist
Superiority in cascade reaction.
(3) activity of the parcel enzyme in high viscosity solution
Enzyme without silk nanometer bead parcel and the enzyme for having silk nanometer bead parcel are placed in the poly- second two of various concentrations
In alcohol (PEG) solution, the relative activity of its enzyme at high viscosities is tested, as a result as shown in Figure 5.It can be seen that
As PEG concentration is raised, solution viscosity becomes larger, and the relative activity of enzyme is gradually lowered, but when PEG concentration is higher than 15v/
V%, that is, be in high viscosity solution, and the speed that the relative activity for having the enzyme of silk nanometer bead parcel declines is significantly lower than without silkworm
The enzyme of silk nanometer bead parcel, illustrate the parcel of silk nanometer bead can protect and be in high viscosity solution in (such as cell liquid, blood
Liquid etc.) enzyme activity, enzyme is under equal conditions preserved activity higher.
Application of the embodiment 3 in antialcoholic drug
Alcohol oxidase enzyme (AOx) and catalase (Cat) are wrapped in silk nanometer bead, a kind of anti-albumen is obtained
The antialcoholic drug of enzyme, its effect is as shown in Figure 6.
In intestinal juice simulated solution, add the enzyme alcohol without silk nanometer bead parcel or have silk nanometer bead parcel
Enzyme alcohol, compares both relative activities, and as shown in Figure 6 a, the enzyme alcohol for having silk nanometer bead parcel is still protected after 2h
Deposit more than 90% activity, and the relative activity of the enzyme after 20 min of the enzyme alcohol without silk nanometer bead parcel quickly falls to
5%.Because during the medicine containing protease reaches intestinal juice after oral, protease present in intestinal juice can promote albumen
The hydrolysis of matter, and the essence of most of enzyme is exactly protein, so as to cause enzyme fast decoupled in intestinal juice to inactivate, and is wrapped up
Silk nanometer bead outside enzyme alcohol can be effectively protected enzyme alcohol and not be decomposed, so that the activity of protective enzyme.Thus may be used
See, silk nanometer bead can effectively prevent the enzyme of its parcel of protease hydrolytic, it is ensured that the performance of enzymatic activity.
In intestinal juice simulated solution, while adding enzyme alcohol and catalase without silk nanometer bead parcel, Huo Zhejia
Enter to have wrapped up the silk nanometer bead of enzyme alcohol and catalase, compare the change of its alcohol concentration over time.Such as
Shown in Fig. 6 b, enzyme alcohol and catalase without silk parcel can not explain alcohol, by after 6h, the concentration of alcohol is still
There is no significant change；And add the enzyme alcohol and catalase that have silk nanometer bead parcel can effectively to degrade alcohol, make
Obtain alcohol concentration to decline rapidly, alcohol concentration is reduced to 3% after 6h, and dispelling effects of alcohol is notable.
As can be seen here, the enzyme that the silk nanometer bead of enzyme alcohol and catalase can effectively protect it to wrap up has been wrapped up
Not by proteases for decomposing, and the alcohol concentration in human body can be efficiently reduced, reach significant dispelling effects of alcohol.
The above, the only present invention preferably specific embodiment, but protection scope of the present invention is not limited thereto,
Any one skilled in the art the invention discloses technical scope in, the change or replacement that can be readily occurred in,
Should all be included within the scope of the present invention.Therefore, protection scope of the present invention should be with scope of the claims
It is defined.
Claims (8)
1. it is a kind of wrap up enzyme silk nanometer bead preparation method, it is characterised in that：Comprise the following steps：
S1, silk solution is prepared, specially：
S11, silk cocoon is shredded, be placed in 5% NaHCO3Boiled in solution, and 30min is stirred with glass bar, repeat the step two
It is secondary；
S12, with gained silk 20min in 60 DEG C of deionized water washing step S11, repeat the step 3 time；
S13, washed silk is placed in 60 DEG C of baking ovens dry, be subsequently solubolized in the LiBr solution of 9.3M, 60 DEG C dissolving 4h；
Gained silk solution two days, changes a water in every 2 hours in S14, the step S13 that dialysed with deionized water；
S15, by step S14 gained silk solution be diluted to 6%w/w；
S2, enzyme is wrapped up into silk, form the silk nanometer bead of parcel enzyme；
Obtained silk nanometer bead is made up of the enzyme core of silk nanometer shell and its parcel, the silk nanometer bead
Particle diameter is 50-150nm.
2. it is according to claim 1 it is a kind of wrap up enzyme silk nanometer bead preparation method, it is characterised in that：Step S2
Specifically include following steps：
S21, silk solution obtained in step S1 is taken, Cord blood 4-8 hours, wherein silkworm in refrigerator are placed in after mixing with enzyme solutions
Silk solution is 5 with the volume ratio of enzyme solutions：1；
S22, gained mixed solution in step S21 is taken, be slowly dropped into a certain amount of acetone with the amount of every drop 25ul, wherein mixing
Solution is 1.2 with the volume ratio of acetone：5, resulting solution 18000rpm is centrifuged 30min；
S23, supernatant is abandoned, add deionized water, supersound washing simultaneously disperses；
S24, by gained mixed solution 18000rpm centrifugations 15min in step S23；
S25, repeat step S23, S24 tri- times, that is, be obtained the silk nanometer bead of parcel enzyme.
3. it is according to claim 2 it is a kind of wrap up enzyme silk nanometer bead preparation method, it is characterised in that：Step
The enzyme class included in enzyme solutions described in S21 is one or more.
4. it is according to claim 3 it is a kind of wrap up enzyme silk nanometer bead preparation method, it is characterised in that：The enzyme
Mass ratio containing glucose oxidase and horseradish peroxidase in solution is 4:1.
5. it is according to claim 2 it is a kind of wrap up enzyme silk nanometer bead preparation method, it is characterised in that：Step
Low temperature described in S21 is 4-10 DEG C.
6. it is according to claim 2 it is a kind of wrap up enzyme silk nanometer bead preparation method, it is characterised in that：Step
Supersound washing described in S23 is specially：2min is shaken after 40% power ultrasonic 30s on the oscillator, is repeated 3-5 times, until dispersion
Uniformly.
7. the application of the silk nanometer bead of a kind of parcel enzyme according to claim any one of 1-6, it is characterised in that：Institute
Stating silk nanometer bead can be in medical domain application.
8. the application of a kind of silk nanometer bead for wrapping up enzyme according to claim 7, it is characterised in that：The silk is received
Nano-sphere can be used to prepare antialcoholic drug, wrap up alcohol hydrogenase and catalase in obtained antialcoholic drug simultaneously.
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| 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 |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201710046253.6A CN106822038B (en) | 2017-01-22 | 2017-01-22 | Preparation method and application of enzyme-coated silk nanospheres |
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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 |
|---|
| FEI WANG,ET AL: "Bioconjugation of Silk Fibroin Nanoparticles with Enzyme and Peptide and Their Characterization", 《ADVANCES IN PROTEIN CHEMISTRY AND STRUCTURAL BIOLOGY》 * |
| SHENZHOU LU,ET AL: "Stabilization of Enzymes in Silk Films", 《BIOMACROMOLECULES》 * |
| 朱祥瑞等: "家蚕丝素固定化过氧化氢酶的制备及其理化特性的研究", 《农业生物技术学报》 * |
| 陈艳芳等: "丝素作为固定化酶载体的研究进展与应用", 《广东蚕业》 * |
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