CN106727341A - A kind of preparation method of antimicrobial form protein peptides nanoparticle complex - Google Patents
A kind of preparation method of antimicrobial form protein peptides nanoparticle complex Download PDFInfo
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Abstract
The present invention provides a kind of preparation method of antimicrobial form protein peptides nanoparticle complex, and operating procedure is：1）The preparation of zinc oxide colloid；2）The preparation of nanometer Zinc material；3）The preparation of protein peptides nanoparticle.Have the beneficial effect that：Nano zine oxide has specific adsorption to act on the yellow crucian carp protein peptides for having antibacterial action, plays the effect of purifying；Yellow crucian carp protein peptides absorption keeps relatively independent space structure and bioactivity, and specific surface area exponentially to increase again, with superpower antibacterial effect on nano zine oxide；Nano material has slight toxicity, and one layer of polypeptide is wrapped up in outer surface, can reduce its toxicity, is conducive to healthy；Preparation method is simple, with low cost, the product good antimicrobial effect of preparation, and will not develop immunity to drugs, and green health is safe, is adapted to large-scale production.
Description
Technical field
The present invention relates to technical field of composite preparation, specifically a kind of antimicrobial form protein peptides-nanoparticle complex
Preparation method.
Background technology
Nano-particle material is also called ultramicro powder material, is made up of nano-particle (nanoparticle).Nano-particle
Also ultramicro powder is, particle of the size between 1~100nm is generally referred to, is to be in the transition that cluster and macro object have a common boundary
Region, from common on from the viewpoint of microcosmic and macroscopical, both atypical microscopic system is also atypical grand for such system
Sight system, is a kind of typical Mesoscopic structure, and it has skin effect, small-size effect and macro quanta tunnel effect.Work as people
Macro object is subdivided into ultramicro powder（Nanoscale）Afterwards, it would indicate that many unusual characteristics, i.e. its optics, heat
Being compared when property in terms of, electricity, magnetics, mechanics and chemistry is with bulk solid will have significant difference.
Nano material has certain uniqueness, when material yardstick it is small to a certain extent when, then must use quantum mechanics instead
The viewpoint of traditional mechanicses is replaced to describe its behavior, when powder particle size is when 10 microns are down to 10 nanometers, though its particle diameter
1000 times are changed into, but will then there are 10 9 powers times when being converted into volume, so in the two behavior that generation is obvious poor
It is different.
The reasons why nano-particle is different from bulk matter is increased its surface area is relative, that is, ultramicron surface cloth
Step structure is expired, this structure represents the unstability atom with high surface energy.This kind of atom is easily adsorbed with foreign atom
Bond, while providing the active atomic of large surface because of particle diameter diminution.
In today that abuse of antibiotics, antibody-resistant bacterium are constantly produced, the research of antibacterial peptide attracts the mesh of domestic and foreign scholars
Light, people are constantly looking for new antibacterial peptide, explore the new mechanism of action of antibacterial peptide.Antibacterial peptide
(antibacterialpeptide, ABP) is widely distributed also known as antimicrobial peptide (antimicrobial peptide, AMP)
It is the important component of biological natural immune defence system in bacterium, viral and various animal and plant body.AMP majorities are one
Quasi-molecule amount is less than 10kDa small-molecular peptides, has multifarious cationic amphiphic peptides in chain length, sequence and structure
(Boman H G.Antibactericidal peptides ：key components needed in immunity[J]
.Cell, 1991,65：205-207 ；Hultmark D.Drosophila immunity ：paths and patterns
[J] .Current Opinion in Immunology, 2003,15：12-19.).Prior art such as Authorization Notice No. is
The Chinese invention patent of CN102051398B, discloses and a kind of prepares antibacterial with pure natural marine products Optimization of Low Value Fish Huang crucian carp as raw material
Peptide, yellow crucian carp albumen antibacterial peptide is rich in essential amino acid, physicochemical property stabilization, with the antibacterial peptide obtained by food source proteolysis
Safe, absorbability is good, good antimicrobial effect, as nutrition and functional form additive or can be developed as drug-fast bacteria in food
Antibiotic.
Prior art such as Authorization Notice No. is the Chinese invention patent of CN103433484B, discloses a kind of bovine serum albumin
In vain-platinum composite nano materials Mimetic enzyme and its preparation method and application, with bovine serum albumin(BSA) as template, by life
Thing mineralization, prepares bovine serum albumin(BSA)-platinum composite nano materials Mimetic enzyme.Its bovine serum albumin(BSA)-platinum
Composite nano materials are prepared from by following methods：Add chloroplatinic acid water-soluble in concentration is Bovine Serum Albumin in Aqueous Solution
Liquid, adds sodium hydrate aqueous solution to obtain mixed solution, heating water bath after mixing；By this solution is by ultrafiltration and washes, ox is obtained
Seralbumin-platinum composite nano materials the aqueous solution；Bovine serum albumin(BSA)-platinum composite nano materials have excellent peroxidating
Enzymatic activity, can catalyzing hydrogen peroxide oxidation 3,3 ', 5, the colour developing of 5 '-tetramethyl biphenyl amine hydrochlorate.The analogue enztme is acidproof simultaneously
Alkali, high temperature resistant, resistance to high salt, with excellent short-term room temperature stability and long-term room-temperature stability.
The content of the invention
Stable in properties it is an object of the invention to provide one kind, specific surface area is big, the superpower antimicrobial form albumen of fungistatic effect
The preparation method of peptide-nanoparticle complex.
The present invention is directed to the problem mentioned in background technology, and the technical scheme taken is：
A kind of preparation method of antimicrobial form protein peptides-nanoparticle complex, specific preparation process：
1）The preparation of zinc oxide colloid：Zinc acetate, ultrasonic dissolution assisting is added to obtain zinc acetate ethanol solution, second in absolute ethyl alcohol
The concentration of sour zinc ethanol solution is 0.08 ~ 0.12mol/L, is carried out in closed container during configuration, prevents ethanol from volatilizing.Stirring,
NaOH ethanol solution is added dropwise in NaOH ethanol solution, NaOH ethanol solution concentration is 0.16 ~ 0.24
Mol/L, NaOH ethanol solution：NaOH volumes of aqueous ethanol ratio is 1:0.8 ~ 1.3, bath temperature is 75 ~ 90 DEG C, is shaken
1.5 ~ 3h of stirring is swung, zinc acetate hydrolysis, colloidal sol, gel obtain zinc oxide colloidal solution；
2）The preparation of nanometer Zinc material：N-hexane and glycol monoethyl ether, n-hexane are added in colloidal solution：Colloidal solution body
Product is than being 1:0.6 ~ 1.5, glycol monoethyl ether：N-hexane volume ratio is 1:100 ~ 500, a small amount of second two is added in n-hexane
Alcohol monomethyl ether can be improved solubility of the ethanol in n-hexane, and the eduction rate of zinc oxide colloid is improved, and improves nanometer
The particle diameter of grain, improves complex properties of product；Stirring, centrifugation is precipitated with absolute ethanol washing, using vacuum dehydrating at lower temperature, very
Reciprocal of duty cycle is 5000 ~ 10000pa, and temperature is 30 ~ 40 DEG C, and drying time is 1.2 ~ 2.9h, obtains nano-zinc oxide powder, is placed in dry
Kept in dark place in dry device；
3）The preparation of protein peptides-nanoparticle：The pH of the yellow crucian carp protein peptides of regulation is 2.5 ~ 3.5, and centrifugation adds nanometer in supernatant
Zinc oxide powder, makes its final concentration of 18 ~ 21mg/mL.Water-bath is vibrated, and bath temperature is 30 ~ 42 DEG C, and 6 ~ 9h is vibrated in water-bath, is turned
Speed is 120 ~ 200rpm.Centrifugation, supernatant is continued to employ, and obtains soluble Huang crucian carp albumen antibacterial peptide-nano zine oxide.Nano zine oxide
Yellow crucian carp protein peptides to there is antibacterial action have specific adsorption to act on, and play the effect of purifying；Yellow crucian carp protein peptides absorption is in nanometer
On zinc oxide, relatively independent space structure and bioactivity, and specific surface area exponentially is kept to increase again, with superpower
Antibacterial effect；Nano material has slight toxicity, and one layer of polypeptide is wrapped up in outer surface, can reduce its toxicity, is conducive to body
Health.
Compared with prior art, the advantage of the invention is that：A small amount of glycol monoethyl ether is added in n-hexane can be made
Solubility of the ethanol in n-hexane is improved, and the eduction rate of zinc oxide colloid is improved；Nano zine oxide is to the Huang that has antibacterial action
Crucian carp protein peptides have specific adsorption to act on, and play the effect of purifying；Yellow crucian carp protein peptides absorption keeps relative on nano zine oxide
Independent space structure and bioactivity, and specific surface area exponentially increases again, with superpower antibacterial effect；Nano material
With slight toxicity, one layer of polypeptide is wrapped up in outer surface, its toxicity can be reduced, be conducive to healthy；Preparation method is simple,
It is with low cost, the product good antimicrobial effect of preparation, and will not develop immunity to drugs, green health is safe, is adapted to extensive raw
Produce.
Brief description of the drawings
Fig. 1 is nano zine oxide transmission electron microscope picture；
Fig. 2 is nano zine oxide XRD analysis collection of illustrative plates；
Fig. 3 is the transmission electron microscope picture of yellow crucian carp albumen antibacterial peptide-nano-zinc oxide compound；
Fig. 4 is the high score rate transmission electron microscope picture of yellow crucian carp albumen antibacterial peptide-nano-zinc oxide compound；
Fig. 5 is the Escherichia coli scanning electron microscope (SEM) photograph without yellow crucian carp albumen antibacterial peptide-nano-zinc oxide compound treatment；
Fig. 6 is the Escherichia coli scanning electron microscope (SEM) photograph that 3h is processed through yellow crucian carp albumen antibacterial peptide-nano-zinc oxide compound.
Specific embodiment
The present invention program is described further below by embodiment：
Embodiment 1：
A kind of preparation method of antimicrobial form protein peptides-nanoparticle complex, specific preparation process：
1）The preparation of zinc oxide colloid：Zinc acetate, ultrasonic dissolution assisting is added to obtain zinc acetate ethanol solution, second in absolute ethyl alcohol
The concentration of sour zinc ethanol solution is 0.08 ~ 0.12mol/L, is carried out in closed container during configuration, prevents ethanol from volatilizing.Stirring,
NaOH ethanol solution is added dropwise in NaOH ethanol solution, NaOH ethanol solution concentration is 0.16 ~ 0.24
Mol/L, NaOH ethanol solution：NaOH volumes of aqueous ethanol ratio is 1:0.8 ~ 1.3, bath temperature is 75 ~ 90 DEG C, is shaken
1.5 ~ 3h of stirring is swung, zinc acetate hydrolysis, colloidal sol, gel obtain zinc oxide colloidal solution；
2）The preparation of nanometer Zinc material：N-hexane and glycol monoethyl ether, n-hexane are added in colloidal solution：Colloidal solution body
Product is than being 1:0.6 ~ 1.5, glycol monoethyl ether：N-hexane volume ratio is 1:100 ~ 500, a small amount of second two is added in n-hexane
Alcohol monomethyl ether can be improved solubility of the ethanol in n-hexane, and the eduction rate of zinc oxide colloid is improved.Stirring, centrifugation, with nothing
Water-ethanol washing precipitation, using vacuum dehydrating at lower temperature, vacuum is 5000 ~ 10000pa, and temperature is 30 ~ 40 DEG C, and drying time is
1.2 ~ 2.9h, obtains nano-zinc oxide powder, is placed in drier and keeps in dark place；
3）The preparation of protein peptides-nanoparticle：The pH of the yellow crucian carp protein peptides of regulation is 2.5 ~ 3.5, and centrifugation adds nanometer in supernatant
Zinc oxide powder, makes its final concentration of 18 ~ 21mg/mL.Water-bath is vibrated, and bath temperature is 30 ~ 42 DEG C, and 6 ~ 9h is vibrated in water-bath, is turned
Speed is 120 ~ 200rpm.Centrifugation, supernatant is continued to employ, and obtains soluble Huang crucian carp albumen antibacterial peptide-nano zine oxide.Nano zine oxide
Yellow crucian carp protein peptides to there is antibacterial action have specific adsorption to act on, and play the effect of purifying；Yellow crucian carp protein peptides absorption is in nanometer
On zinc oxide, relatively independent space structure and bioactivity, and specific surface area exponentially is kept to increase again, with superpower
Antibacterial effect；Nano material has slight toxicity, and one layer of polypeptide is wrapped up in outer surface, can reduce its toxicity, is conducive to body
Health.
Embodiment 2：
A kind of preparation method of antimicrobial form protein peptides-nanoparticle complex, most preferably preparation process：
1）The preparation of zinc oxide colloid：Zinc acetate, ultrasonic dissolution assisting is added to obtain zinc acetate ethanol solution, second in absolute ethyl alcohol
The concentration of sour zinc ethanol solution is 0.1mol/L, is carried out in closed container during configuration, prevents ethanol from volatilizing.Stirring, by hydrogen-oxygen
Change sodium ethoxide solution to be added dropwise in NaOH ethanol solution, NaOH ethanol solution concentration is 0.2mol/L, hydroxide
Sodium ethoxide solution：NaOH volumes of aqueous ethanol ratio is 1:1, bath temperature is 80 DEG C, vibration stirring 2h, zinc acetate hydrolysis,
Colloidal sol, gel obtain zinc oxide colloidal solution；
2）The preparation of nanometer Zinc material：N-hexane and glycol monoethyl ether, n-hexane are added in colloidal solution：Colloidal solution body
Product is than being 1:1, glycol monoethyl ether：N-hexane volume ratio is 1:200, a small amount of glycol monoethyl ether energy is added in n-hexane
Improve solubility of the ethanol in n-hexane, the eduction rate of zinc oxide colloid is improved.Stirring, centrifugation, uses absolute ethanol washing
Precipitation, using vacuum dehydrating at lower temperature, vacuum is 7000pa, and temperature is 35 DEG C, and drying time is 2.3h, obtains nano zine oxide
Powder, is placed in drier and keeps in dark place.There is Fig. 1 and Fig. 2 to understand the chemical composition and structure of nano zine oxide；
3）The preparation of protein peptides-nanoparticle：The pH of the yellow crucian carp protein peptides of regulation is 3.0, and centrifugation is added nano oxidized in supernatant
Zinc powder, makes its final concentration of 19.78mg/mL.Water-bath is vibrated, and bath temperature is 37 DEG C, water-bath vibration 6.9h, and rotating speed is
160rpm.Centrifugation, supernatant is continued to employ, and obtains soluble Huang crucian carp albumen antibacterial peptide-nano zine oxide.Antibacterial is understood by Fig. 3 and Fig. 4
Type Huang crucian carp protein peptides are adsorbed on nano-titanium oxide well.Nano zine oxide has specifically to the yellow crucian carp protein peptides for having antibacterial action
Property suction-operated, plays the effect of purifying.Can be seen that yellow crucian carp protein peptides-nano combined physical efficiency makes large intestine bar by Fig. 5 and Fig. 6 contrasts
Bacterium surface folding, makes it lose activity.The absorption of yellow crucian carp protein peptides on nano zine oxide, keep relatively independent space structure and
Bioactivity, and specific surface area exponentially increases again, with superpower antibacterial effect；Nano material has slight toxicity,
One layer of polypeptide is wrapped up in outer surface, its toxicity can be reduced, be conducive to healthy.
Routine operation in operating procedure of the invention is well known to those skilled in the art, and is not repeated herein.
Glycol monoethyl ether in step 2：The volume ratio of n-hexane is not limited only to 1:100 ~ 500, also including 1:102 or 1:
104 or 1:106 ... ..., or 1:496 or 1:498 or 1:500.
Embodiment described above has been described in detail to technical scheme, it should be understood that the above is only
It is specific embodiment of the invention, is not intended to limit the invention, all any modifications made in spirit of the invention,
Supplement or similar fashion replacement etc., should be included within the scope of the present invention.
Claims (9)
1. a kind of preparation method of antimicrobial form protein peptides-nanoparticle complex, it is characterised in that following steps：
1）The preparation of zinc oxide colloid：Zinc acetate is added in absolute ethyl alcohol, zinc acetate ethanol solution is obtained, stirred, add hydrogen
Oxidation sodium ethoxide solution, water-bath vibration, obtains zinc oxide colloidal solution；
2）The preparation of nanometer Zinc material：N-hexane and glycol monoethyl ether are added in colloidal solution, centrifugation, washing precipitation is done
It is dry, obtain nano-zinc oxide powder；
3）The preparation of protein peptides-nanoparticle：The pH of the yellow crucian carp protein peptides of regulation, centrifugation adds described nano oxygen in supernatant
Change zinc powder, water-bath vibration, centrifugation, supernatant is continued to employ, obtain soluble Huang crucian carp albumen antibacterial peptide-nano zine oxide.
2. the preparation method of a kind of antimicrobial form protein peptides-nanoparticle complex according to claim 1, it is characterised in that：
The concentration of zinc acetate ethanol solution is 0.08 ~ 0.12mol/L in described step 1, is carried out in closed container during configuration, hydrogen-oxygen
It is 0.16 ~ 0.24 mol/L to change sodium ethoxide solution concentration.
3. the preparation method of a kind of antimicrobial form protein peptides-nanoparticle complex according to claim 1, it is characterised in that：
In described step 1 ultrasonic dissolution assisting is used in zinc acetate ethanol solution and/or NaOH ethanol solution configuration process.
4. the preparation method of a kind of antimicrobial form protein peptides-nanoparticle complex according to claim 1, it is characterised in that：
NaOH ethanol solution is added dropwise in NaOH ethanol solution in described step 1.
5. the preparation method of a kind of antimicrobial form protein peptides-nanoparticle complex according to claim 1, it is characterised in that：
NaOH ethanol solution in described step 1：NaOH volumes of aqueous ethanol ratio is 1:0.8 ~ 1.3, bath temperature be 75 ~
90 DEG C, 1.5 ~ 3h of vibration stirring.
6. the preparation method of a kind of antimicrobial form protein peptides-nanoparticle complex according to claim 1, it is characterised in that：
N-hexane in described step 2：Colloidal solution volume ratio is 1:0.6 ~ 1.5, glycol monoethyl ether：N-hexane volume ratio is 1:
100 ~ 500, the Nano-Zinc powder that will be obtained is placed in drier and keeps in dark place.
7. the preparation method of a kind of antimicrobial form protein peptides-nanoparticle complex according to claim 1, it is characterised in that：
Vacuum dehydrating at lower temperature is used in described step 2, vacuum is 5000 ~ 10000pa, and temperature is 30 ~ 40 DEG C, and drying time is
1.2~2.9h。
8. the preparation method of a kind of antimicrobial form protein peptides-nanoparticle complex according to claim 1, it is characterised in that：
The pH that yellow crucian carp protein peptides are adjusted in described step 3 is 2.5 ~ 3.5, adds nano zine oxide in supernatant, final concentration of 18 ~
21mg/mL。
9. the preparation method of a kind of antimicrobial form protein peptides-nanoparticle complex according to claim 1, it is characterised in that：
Bath temperature is 30 ~ 42 DEG C in described step 3, and 6 ~ 9h is vibrated in water-bath, and rotating speed is 120 ~ 200rpm.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107034260A (en) * | 2017-06-27 | 2017-08-11 | 钦州学院 | A kind of preparation method of Nano-Zinc ostreae testa pulverata |
| KR102137188B1 (en) * | 2020-03-20 | 2020-08-28 | 유피씨(주) | Antibacterial and Deodorizing Composition, Method for preparing a composition having antibacterial and deodorizing function and Fabric using the same |
| CN115281284A (en) * | 2022-09-26 | 2022-11-04 | 北京中科泰训科技有限公司 | Antibacterial peptide composite nano-particles, preparation method and application of antibacterial peptide composite nano-particles as feed additive |
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| CN102051398A (en) * | 2009-10-29 | 2011-05-11 | 浙江海洋学院 | Method for preparing half-fin anchovy antibacterial peptides |
| CN104667253A (en) * | 2015-02-02 | 2015-06-03 | 深圳市大鳄生物科技股份有限公司 | Crocodile nano-polypeptide and preparation method thereof |
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| CN102051398A (en) * | 2009-10-29 | 2011-05-11 | 浙江海洋学院 | Method for preparing half-fin anchovy antibacterial peptides |
| CN104667253A (en) * | 2015-02-02 | 2015-06-03 | 深圳市大鳄生物科技股份有限公司 | Crocodile nano-polypeptide and preparation method thereof |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107034260A (en) * | 2017-06-27 | 2017-08-11 | 钦州学院 | A kind of preparation method of Nano-Zinc ostreae testa pulverata |
| CN107034260B (en) * | 2017-06-27 | 2021-07-02 | 钦州学院 | Preparation method of nano-zinc oyster powder |
| KR102137188B1 (en) * | 2020-03-20 | 2020-08-28 | 유피씨(주) | Antibacterial and Deodorizing Composition, Method for preparing a composition having antibacterial and deodorizing function and Fabric using the same |
| CN115281284A (en) * | 2022-09-26 | 2022-11-04 | 北京中科泰训科技有限公司 | Antibacterial peptide composite nano-particles, preparation method and application of antibacterial peptide composite nano-particles as feed additive |
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