CN110373153A - A kind of bioadhesive of high adhesive strength and preparation method thereof - Google Patents
A kind of bioadhesive of high adhesive strength and preparation method thereof Download PDFInfo
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- CN110373153A CN110373153A CN201910777873.6A CN201910777873A CN110373153A CN 110373153 A CN110373153 A CN 110373153A CN 201910777873 A CN201910777873 A CN 201910777873A CN 110373153 A CN110373153 A CN 110373153A
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- Prior art keywords
- super
- bioadhesive
- surfactant
- albumen
- charge
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09J—ADHESIVES; NON-MECHANICAL ASPECTS OF ADHESIVE PROCESSES IN GENERAL; ADHESIVE PROCESSES NOT PROVIDED FOR ELSEWHERE; USE OF MATERIALS AS ADHESIVES
- C09J11/00—Features of adhesives not provided for in group C09J9/00, e.g. additives
- C09J11/02—Non-macromolecular additives
- C09J11/06—Non-macromolecular additives organic
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09J—ADHESIVES; NON-MECHANICAL ASPECTS OF ADHESIVE PROCESSES IN GENERAL; ADHESIVE PROCESSES NOT PROVIDED FOR ELSEWHERE; USE OF MATERIALS AS ADHESIVES
- C09J189/00—Adhesives based on proteins; Adhesives based on derivatives thereof
Abstract
The present invention provides bioadhesives of a kind of high adhesive strength and preparation method thereof, are made of super charge albumen and surfactant；The surfactant is cationic surface active agent or anionic azobenzene surfactant；The cationic surface active agent has catechol structural unit.The present invention is acted on using electrostatic force, π-cation and the pi-pi accumulation between super charge albumen and surfactant, improves the adhesion property of bioprotein adhesive.The experimental results showed that the bioadhesive in the present invention shows excellent bond properties in steel disc, polyethylene and polyvinyl chloride surface.Also, preparation process of the present invention is simple, is not necessarily to special installation, is at low cost and environmentally protective.Compared with prior art, the present invention has good biocompatibility and degradability.
Description
Technical field
The invention belongs to the bioadhesives and its preparation of technical field of bioengineering more particularly to a kind of high adhesive strength
Method.
Background technique
Adhesive is a kind of under prescribed conditions, making the material of two or more surface bondings together.It is in height
It is played an important role in technology and field of biomedicine.It equally, in nature, is raw across the adhesion of multiple length dimensions
The necessary condition deposited.
By the inspiration of nature, researcher by the research to the relevant adhesive of biologic artifact and mechanism of attachment,
The adhesives of artificial synthesized this Adhesion properties of simulation various out.But these adhesives usually require a large amount of time, material resources,
Financial resources are artificial synthesized to carry out, and these adhesives also need polymerization reaction or realize gluing by ultraviolet irradiation, and
And it is easy to produce toxic side effect, therefore the application which greatly limits existing adhesive on the fields such as medicine and industry.
Therefore, in the science and technology of adhesive, the jointing material of high bonding characteristic and good biological compatibility is needed, with
Just prepared adhesive is preferably applied to more fields.
Summary of the invention
The purpose of the present invention is to provide bioadhesive of a kind of high adhesive strength and preparation method thereof, in the present invention
Bioadhesive has high-adhesion and good bio-compatibility simultaneously.
The present invention provides a kind of bioadhesive of high adhesive strength, is made of super charge albumen and surfactant；
The surfactant is cationic surface active agent or anionic azobenzene surfactant；
The cationic surface active agent has catechol structural unit.
Preferably, the super charge albumen follows the steps below expression:
Super positive charge albumen K series or super negative electrical charge albumen E serial carrier plasmid are transformed into Pichia pastoris respectively
(GS115/X-33/KM71) or E. coli expression strains (BL21/BL21DE3/BLRDE3), picking monoclonal colonies use LB
Culture solution is incubated overnight；TB culture medium is added in the expression seed liquor being activated overnight, reaching OD600 numerical value to bacterium solution is
When 0.8, inducer isopropylthio-β-D- Thiogalactopyranoside is added and cools to 30 degree of progress overexpressions.It induces 12 small
When collect thallus, be resuspended with lysis buffer, protease inhibitors be added, DNA enzymatic, lysozyme is crushed bacterium with high pressure crusher
Body more than 10000rpm revolving speed is centrifuged, and collects supernatant, and HPLC purifying finally obtains super positive charge albumen K or super negative electricity
Lotus albumen E.
Preferably, the super charge albumen is super positive charge albumen or super negative electrical charge albumen；
The bioadhesive is made of super positive charge albumen and anionic azobenzene surfactant；Alternatively,
The bioadhesive is made of super negative electrical charge albumen and cationic surface active agent.
Preferably, the charge number of the super positive charge albumen is 72~108；
The charge number of the super negative electrical charge albumen is 36~144.
Preferably, the cationic surface active agent is C19H33ClN2O3。
Preferably, the anion azobenzene surfactant is C19H23N2NaO7S。
Preferably, the molar ratio of the super charge albumen and surfactant is 1:(1~5).
Preferably, the water content of the bioadhesive is 5~15%.
The present invention provides a kind of preparation method of the bioadhesive of high adhesive strength, comprising the following steps:
A super charge albumen and surfactant are dissolved in ultrapure water respectively), obtain super charge protein solution and table
Face activator solution；
The surfactant is cationic surface active agent or anionic azobenzene surfactant；
The cationic surface active agent has catechol structural unit；
B) surfactant solution is added dropwise in super charge protein solution, is uniformly mixed, it is molten to obtain adhesive
Liquid；
C it) will be lyophilized 0.2~0.5 hour after the centrifugation of described adhesive solution, obtain the bioadhesive of high adhesive strength.
Preferably, the rate of the centrifugation is 15000~18000rpm；
The time of the centrifugation is 5~20min.
The present invention provides a kind of bioadhesives of high adhesive strength, by super charge albumen and surfactant group
At；The surfactant is cationic surface active agent or anionic azobenzene surfactant；The cation
Type surfactant has catechol structural unit.The present invention utilizes the electrostatic between super charge albumen and surfactant
Power effect, π-cation and pi-pi accumulation effect, improve the adhesion property of bioprotein adhesive.The experimental results showed that the present invention
In bioadhesive excellent bond properties is shown in steel disc, polyethylene and polyvinyl chloride surface.Also, system of the present invention
Standby simple process is not necessarily to special installation, is at low cost and environmentally protective.Compared with prior art, the present invention has good biology
Compatibility and degradability.
Detailed description of the invention
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technical description to be briefly described, it should be apparent that, the accompanying drawings in the following description is only this
The embodiment of invention for those of ordinary skill in the art without creative efforts, can also basis
The attached drawing of offer obtains other attached drawings.
Fig. 1 is the protein binder shearing force strength test stretching device schematic diagram in the present invention；
Fig. 2 is the weighing test chart of bioadhesive in the embodiment of the present invention 2；
Shearing force intensity of the Fig. 3 for bioadhesive in the embodiment of the present invention 1~2 on unlike material surface；
Shearing force intensity of the Fig. 4 for bioadhesive in the embodiment of the present invention 3~5 on unlike material surface.
Specific embodiment
The present invention provides a kind of bioadhesives of high adhesive strength, by super charge albumen and surfactant group
At；
The surfactant is cationic surface active agent or anionic azobenzene surfactant；
The cationic surface active agent has catechol structural unit.
In the present invention, the super charge albumen is unfolded super charge albumen；The super charge albumen can
To be super positive charge albumen (K), it is also possible to super negative electrical charge albumen (E), when using super positive charge albumen, the surface
Activating agent is preferably anionic azobenzene surfactant；When using super negative electrical charge albumen, the surfactant is excellent
It is selected as cationic surface active agent.
In the present invention, the super charge albumen is preferably prepared according to the following steps to obtain:
Super positive charge albumen K series or super negative electrical charge albumen E serial carrier plasmid are transformed into Pichia pastoris respectively
(GS115/X-33/KM71) or E. coli expression strains (BL21/BL21DE3/BLRD E3), picking monoclonal colonies use LB
Culture solution is incubated overnight；TB culture medium is added in the expression seed liquor being activated overnight, reaching OD600 numerical value to bacterium solution is
When 0.8, inducer isopropylthio-β-D- Thiogalactopyranoside is added and cools to 30 degree of progress overexpressions.It induces 12 small
When collect thallus, be resuspended with lysis buffer, protease inhibitors be added, DNA enzymatic, lysozyme is crushed bacterium with high pressure crusher
Body more than 10000rpm revolving speed is centrifuged, and collects supernatant, and HPLC purifying finally obtains super positive charge albumen K or super negative electricity
Lotus albumen E.
In the present invention, the charge number preferably 72~108 of the super positive charge albumen, the super negative electrical charge albumen
Charge number is 36~144.
The cationic surface active agent (PDD) preferably has catechol structural unit, more preferably
C19H33ClN2O3, specific structure is shown in formula I；
The anionic azobenzene surfactant (NAT) is preferably C19H33ClN2O3；Specific structure such as Formula II institute
Show；
In the present invention, the super charge albumen (in terms of lysine) and the molar ratio of the surfactant are preferably
1:(1~5), more preferably 1:(2~4), most preferably 1:1.
The water content of bioadhesive in the present invention is preferably 5~15%, and more preferably 8~10%.
The present invention also provides a kind of preparation methods of the bioadhesive of high adhesive strength, comprising the following steps:
A super charge albumen and surfactant are dissolved in ultrapure water respectively), obtain super charge protein solution and table
Face activator solution；
The surfactant is cationic surface active agent or anionic azobenzene surfactant；
The cationic surface active agent has catechol structural unit；
B) surfactant solution is added dropwise in super charge protein solution, is uniformly mixed, it is molten to obtain adhesive
Liquid；
C it) will be lyophilized 0.2~0.5 hour after the centrifugation of described adhesive solution, obtain the bioadhesive of high adhesive strength.
In the present invention, the super charge albumen, cationic surface active agent and anionic azobenzene surface
The type of activating agent, dosage and even from grade charge albumen described above, cationic surface active agent and anionic
Type, the dosage of pyridine class surfactant are consistent with source, and details are not described herein.
In the present invention, the molar concentration of the super charge protein solution is preferably 200~300 μm of ol/L, more preferably
For 220~250 μm of ol/L；The molar concentration of the surfactant solution is preferably 10~20 μm of ol/L.
After obtaining binder solution, described adhesive solution is carried out high speed centrifugation by the present invention, is then directly lyophilized, is obtained
The bioadhesive of high adhesive strength.
In the present invention, the rate of the centrifugation is preferably 15000~18000rpm, more preferably 16000~
17000rpm, most preferably 16800rpm；The time of the centrifugation is preferably 5~20min, more preferably 10~15min.
The time of the freeze-drying is preferably 0.2~0.5 hour.
Bioadhesive in the present invention can be applied to steel disc surface, aluminum flake surface, polyethylene surface and polyvinyl chloride
Surface shows good Adhesion property.
The present invention provides a kind of bioadhesives of high adhesive strength, by super charge albumen and surfactant group
At；The surfactant is cationic surface active agent or anionic azobenzene surfactant；The cation
Type surfactant has catechol structural unit.The present invention utilizes the electrostatic between super charge albumen and surfactant
Power effect, π-cation and pi-pi accumulation effect, improve the adhesion property of bioprotein adhesive.The experimental results showed that the present invention
In bioadhesive excellent bond properties is shown in steel disc, polyethylene and polyvinyl chloride surface.Also, system of the present invention
Standby simple process is not necessarily to special installation, is at low cost and environmentally protective.Compared with prior art, the present invention has good biology
Compatibility and degradability.
In order to further illustrate the present invention, with reference to embodiments to a kind of biology of high adhesive strength provided by the invention
Adhesive and preparation method thereof is described in detail, but cannot be understood as limiting the scope of the present invention.
Embodiment 1
Super positive charge albumen K serial carrier plasmid is transformed into E. coli expression strains BL21DE3, picking list respectively
Colonies are incubated overnight with LB culture solution；TB culture medium is added in the expression seed liquor being activated overnight, is reached to bacterium solution
When OD600 numerical value is 0.8, inducer isopropylthio-β-D- Thiogalactopyranoside is added and cools to 30 degree and carried out scale
It reaches.12 hours collection thallus are induced, are resuspended with lysis buffer, protease inhibitors, DNA enzymatic is added, lysozyme is crushed with height
Broken crusher machine thallus more than 10000rpm revolving speed is centrifuged, and collects supernatant, and HPLC purifying finally obtains super positive charge albumen
K.The super positive charge protein band quantity of electric charge of this case study on implementation is 72.
Compound concentration is the unfolded positive charge protein solution of 220 μm of ol/L and the azobenzene of 10 μm of ol/L respectively
Aqueous surfactant solution.By both mixing, high speed centrifugation 10 minutes at 16800rpm after five minutes are shaken.Abandon upper liquid
It is lyophilized 0.2 hour afterwards, bioadhesive (K72-NAT) can be obtained.
Embodiment 2
Bioadhesive (K108-NAT) is prepared according to the method in embodiment 1, unlike, in the present embodiment
The charge number of super positive charge albumen is 108.
Bioadhesive in the present embodiment is applied to the bottom of two 20mL vials, is bonded together,
Weighing test is carried out, as a result as shown in Fig. 2, having Fig. 2 it is found that the bioadhesive in the present embodiment can be held for removing surface
By the weight of 600g.
Embodiment 3
Super negative electrical charge albumen E serial carrier plasmid is transformed into E. coli expression strains BL21DE3, picking list respectively
Colonies are incubated overnight with LB culture solution；TB culture medium is added in the expression seed liquor being activated overnight, is reached to bacterium solution
When OD600 numerical value is 0.8, inducer isopropylthio-β-D- Thiogalactopyranoside is added and cools to 30 degree and carried out scale
It reaches.12 hours collection thallus are induced, are resuspended with lysis buffer, protease inhibitors, DNA enzymatic is added, lysozyme is crushed with height
Broken crusher machine thallus more than 10000rpm revolving speed is centrifuged, and collects supernatant, and HPLC purifying finally obtains super negative electrical charge albumen
E.The each molecule charge number of E series is 36 in this case study on implementation.
Compound concentration is the unfolded negative electrical charge protein solution of 220 μm of ol/L and the cation form of 10 μm of ol/L respectively
Face aqueous surfactant solutions.By both mixing, high speed centrifugation 10 minutes at 16800rpm after five minutes are shaken.After abandoning upper liquid
Freeze-drying 0.2 hour, can be obtained bioadhesive (E36-PDD).
Embodiment 4
Bioadhesive (E72-PDD) is prepared according to the preparation method in embodiment 3, unlike, the present embodiment
In super negative electrical charge albumen charge number be 72.
Embodiment 5
Bioadhesive (E14-PDD) is prepared according to the preparation method in embodiment 3, unlike, the present embodiment
In super negative electrical charge albumen charge number be 144.
Bioadhesive in Examples 1 to 2 is subjected to bond properties test on unlike material surface, as a result such as Fig. 3 institute
Show, shearing force intensity of the Fig. 3 for bioadhesive in the embodiment of the present invention 1~2 on unlike material surface.The adhesive is in difference
Superpower adhesion property is shown (including steel disc, aluminium flake, polyethylene and polyvinyl chloride) on egative film.
Bioadhesive in embodiment 3~5 is subjected to bond properties test on unlike material surface, as a result such as Fig. 4 institute
Show, shearing force intensity of the Fig. 4 for bioadhesive in the embodiment of the present invention 3~5 on unlike material surface.The adhesive is in difference
(including steel disc, aluminium flake, polyethylene and polyvinyl chloride) shows good adhesion property on egative film.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of bioadhesive of high adhesive strength is made of super charge albumen and surfactant；
The surfactant is cationic surface active agent or anionic azobenzene surfactant；
The cationic surface active agent has catechol structural unit.
2. bioadhesive according to claim 1, which is characterized in that the super charge albumen according to the following steps into
Row expression:
Super positive charge albumen K series or super negative electrical charge albumen E serial carrier plasmid are transformed into Pichia pastoris or big respectively
Enterobacteria expresses bacterial strain, and picking monoclonal colonies are incubated overnight with LB culture solution；The expression seed liquor being activated overnight is added
Enter TB culture medium, when it is 0.8 that bacterium solution, which reaches OD600 numerical value, inducer isopropylthio-β-D- Thiogalactopyranoside is added simultaneously
Cool to 30 degree of progress overexpressions；12 hours collection thallus are induced, are resuspended with lysis buffer, protease inhibitors is added,
DNA enzymatic, lysozyme are crushed thallus with high pressure crusher, more than 10000rpm revolving speed are centrifuged, and collect supernatant, and HPLC is purified, most
Super positive charge albumen K or super negative electrical charge albumen E is obtained eventually.
3. bioadhesive according to claim 1, which is characterized in that the super charge albumen is super positive charge egg
White or super negative electrical charge albumen；
The bioadhesive is made of super positive charge albumen and anionic azobenzene surfactant；Alternatively, described
Bioadhesive is made of super negative electrical charge albumen and cationic surface active agent.
4. bioadhesive according to claim 1, which is characterized in that the charge number of the super positive charge albumen is 72
~108；
The charge number of the super negative electrical charge albumen is 36~144.
5. bioadhesive according to claim 1, which is characterized in that the cationic surface active agent is
C19H33ClN2O3。
6. bioadhesive according to claim 1, which is characterized in that the anion azobenzene surfactant is
C19H23N2NaO7S。
7. bioadhesive according to claim 1, which is characterized in that the super charge albumen and surfactant
Molar ratio is 1:(1~5).
8. bioadhesive according to claim 1, which is characterized in that the water content of the bioadhesive be 5~
15%.
9. a kind of preparation method of the bioadhesive of high adhesive strength, comprising the following steps:
A super charge albumen and surfactant are dissolved in ultrapure water respectively), super charge protein solution is obtained and surface is living
Property agent solution；
The surfactant is cationic surface active agent or anionic azobenzene surfactant；
The cationic surface active agent has catechol structural unit；
B) surfactant solution is added dropwise in super charge protein solution, is uniformly mixed, obtains binder solution；
C it) will be lyophilized 0.2~0.5 hour after the centrifugation of described adhesive solution, obtain the bioadhesive of high adhesive strength.
10. preparation method according to claim 9, which is characterized in that the rate of the centrifugation be 15000~
18000rpm；
The time of the centrifugation is 5~20min.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113769155A (en) * | 2021-08-12 | 2021-12-10 | 中国人民解放军海军军医大学 | Biological adhesive based on biological engineering protein and preparation method thereof |
| CN114292621A (en) * | 2021-12-29 | 2022-04-08 | 清华大学 | High-low temperature adhesive based on bioengineering protein and preparation method thereof |
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| CN106636016A (en) * | 2017-02-24 | 2017-05-10 | 天津大学 | Method for assisting self-assembly of virus-like particles by introducing positive and negative charges and application |
| WO2019059504A1 (en) * | 2017-09-20 | 2019-03-28 | 주식회사 포스코 | Coacervate formed from catechol-substituted anionic polymer, adhesive comprising same, and method for producing same |
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| JP2004131741A (en) * | 2004-01-20 | 2004-04-30 | Nippi Kollagen Kogyo Kk | Composition for sausage casing, containing adhesive component |
| CN101978040A (en) * | 2008-01-24 | 2011-02-16 | 犹他卅大学研究基金会 | Adhesive complex coacervates and methods of making and using thereof |
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| CN113769155A (en) * | 2021-08-12 | 2021-12-10 | 中国人民解放军海军军医大学 | Biological adhesive based on biological engineering protein and preparation method thereof |
| CN114292621A (en) * | 2021-12-29 | 2022-04-08 | 清华大学 | High-low temperature adhesive based on bioengineering protein and preparation method thereof |
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