CN110373153B - Biological adhesive with high adhesive strength and preparation method thereof - Google Patents
Biological adhesive with high adhesive strength and preparation method thereof Download PDFInfo
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- CN110373153B CN110373153B CN201910777873.6A CN201910777873A CN110373153B CN 110373153 B CN110373153 B CN 110373153B CN 201910777873 A CN201910777873 A CN 201910777873A CN 110373153 B CN110373153 B CN 110373153B
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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
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Abstract
The invention provides a biological adhesive with high adhesive strength and a preparation method thereof, which consists of super charge protein and a surfactant; the surfactant is a cationic surfactant or an anionic azobenzene surfactant; the cationic surfactant has a catechol structural unit. The invention utilizes the electrostatic force effect, the pi-cation and the pi-pi accumulation effect between the super charge protein and the surfactant to improve the adhesion performance of the biological protein adhesive. Experimental results show that the biological adhesive disclosed by the invention has excellent adhesive performance on the surfaces of steel sheets, polyethylene and polyvinyl chloride. In addition, the preparation method is simple in preparation process, free of special equipment, low in cost and environment-friendly. Compared with the prior art, the invention has good biocompatibility and degradability.
Description
Technical Field
The invention belongs to the technical field of bioengineering, and particularly relates to a biological adhesive with high adhesive strength and a preparation method thereof.
Background
An adhesive is a material used to bond two or more surfaces together under specified conditions. It plays an important role in the high technology and biomedical fields. Also, in nature, adhesions spanning multiple length scales are a necessary condition for survival.
Inspired by nature, researchers have artificially synthesized various adhesives that mimic this adhesion performance through studies of adhesives and adhesion mechanisms associated with biological organisms. However, these adhesives usually require a lot of time, material and financial resources for artificial synthesis, and they also require polymerization or adhesion by ultraviolet irradiation and are liable to cause toxic side effects, thus greatly limiting the applications of the existing adhesives in the fields of medicine and industry.
Therefore, in the scientific art of adhesives, there is a need for adhesive materials with high bonding properties and good biocompatibility in order to better apply the prepared adhesives to more fields.
Disclosure of Invention
The invention aims to provide a biological adhesive with high adhesive strength and a preparation method thereof.
The invention provides a biological adhesive with high adhesive strength, which consists of super charge protein and a surfactant;
the surfactant is a cationic surfactant or an anionic azobenzene surfactant;
the cationic surfactant has a catechol structural unit.
Preferably, the supercharged protein is expressed according to the following steps:
respectively transforming the super positive charge protein K series or super negative charge protein E series carrier plasmids into pichia pastoris (GS115/X-33/KM71) or escherichia coli expression strains (BL21/BL21DE3/BLRDE3), selecting a monoclonal colony, and carrying out overnight culture by using LB culture solution; adding the overnight activated expression seed liquid into a TB culture medium, adding an inducer isopropyl-beta-D-thiogalactopyranoside when the bacterial liquid reaches an OD600 value of 0.8, and cooling to 30 ℃ for overexpression. Inducing for 12 hours, collecting thalli, resuspending with lysis buffer solution, adding protease inhibitor, DNA enzyme and lysozyme, crushing thalli with a high-pressure crusher, centrifuging at a rotating speed of more than 10000rpm, collecting supernatant, and purifying by HPLC (high performance liquid chromatography) to finally obtain the super positive charge protein K or the super negative charge protein E.
Preferably, the super-charged protein is a super-positive charged protein or a super-negative charged protein;
the biological adhesive consists of super positive charge protein and anionic azobenzene surfactant; alternatively, the bioadhesive is comprised of a super negatively charged protein and a cationic surfactant.
Preferably, the charge number of the super positive charge protein is 72-108;
the charge number of the super negative charge protein is 36-144.
Preferably, the cationic surfactant is C19H33ClN2O3。
Preferably, the anionic azobenzene surfactant is C19H23N2NaO7S。
Preferably, the molar ratio of the super charged protein to the surfactant is 1: (1-5).
Preferably, the water content of the biological adhesive is 5-15%.
The invention provides a preparation method of a biological adhesive with high adhesive strength, which comprises the following steps:
A) respectively dissolving super-charge protein and a surfactant in ultrapure water to obtain a super-charge protein solution and a surfactant solution;
the surfactant is a cationic surfactant or an anionic azobenzene surfactant;
the cationic surfactant has a catechol structural unit;
B) dropwise adding the surfactant solution into the super-charge protein solution, and uniformly mixing to obtain an adhesive solution;
C) and centrifuging the adhesive solution, and freeze-drying for 0.2-0.5 h to obtain the biological adhesive with high adhesive strength.
Preferably, the speed of the centrifugation is 15000-18000 rpm;
the centrifugation time is 5-20 min.
The invention provides a biological adhesive with high adhesive strength, which consists of super-charged protein and a surfactant; the surfactant is a cationic surfactant or an anionic azobenzene surfactant; the cationic surfactant has a catechol structural unit. The invention utilizes the electrostatic force effect, the pi-cation and the pi-pi accumulation effect between the super charge protein and the surfactant to improve the adhesion performance of the biological protein adhesive. Experimental results show that the biological adhesive disclosed by the invention has excellent adhesive performance on the surfaces of steel sheets, polyethylene and polyvinyl chloride. In addition, the preparation method is simple in preparation process, free of special equipment, low in cost and environment-friendly. Compared with the prior art, the invention has good biocompatibility and degradability.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the provided drawings without creative efforts.
FIG. 1 is a schematic view of a tensile apparatus for testing the shear strength of a protein adhesive in the present invention;
FIG. 2 is a graph showing a weighing test of the bioadhesive in example 2 of the present invention;
FIG. 3 shows the shear strength of the bio-adhesive on the surface of different materials in examples 1-2 of the present invention;
FIG. 4 shows the shear strength of the bio-adhesive on the surface of different materials in examples 3 to 5 of the present invention.
Detailed Description
The invention provides a biological adhesive with high adhesive strength, which consists of super-charged protein and a surfactant;
the surfactant is a cationic surfactant or an anionic azobenzene surfactant;
the cationic surfactant has a catechol structural unit.
In the present invention, the supercharged protein is an unfolded supercharged protein; the super charge protein can be super positive charge protein (K) or super negative charge protein (E), and when the super positive charge protein is used, the surfactant is preferably anionic azobenzene surfactant; when a super negatively charged protein is used, the surfactant is preferably a cationic surfactant.
In the present invention, the supercharged protein is preferably prepared by the following steps:
respectively transforming the super positive charge protein K series or super negative charge protein E series carrier plasmids into pichia pastoris (GS115/X-33/KM71) or escherichia coli expression strains (BL21/BL21DE3/BLRD E3), selecting a monoclonal colony, and carrying out overnight culture by using LB culture solution; adding the overnight activated expression seed liquid into a TB culture medium, adding an inducer isopropyl-beta-D-thiogalactopyranoside when the bacterial liquid reaches an OD600 value of 0.8, and cooling to 30 ℃ for overexpression. Inducing for 12 hours, collecting thalli, resuspending with lysis buffer solution, adding protease inhibitor, DNA enzyme and lysozyme, crushing thalli with a high-pressure crusher, centrifuging at a rotating speed of more than 10000rpm, collecting supernatant, and purifying by HPLC (high performance liquid chromatography) to finally obtain the super positive charge protein K or the super negative charge protein E.
In the invention, the charge number of the super positive charge protein is preferably 72-108, and the charge number of the super negative charge protein is 36-144.
The cationic surfactant (PDD) preferably has catechol building blocks, more preferably C19H33ClN2O3The specific structure is shown as formula I;
the anionic azobenzene surfactant (NAT) is preferably C19H33ClN2O3(ii) a The specific structure is shown as formula II;
in the present invention, the molar ratio of the supercharged protein (in lysine) to the surfactant is preferably 1: (1-5), more preferably 1: (2-4), most preferably 1: 1.
The water content of the biological adhesive is preferably 5-15%, and more preferably 8-10%.
The invention also provides a preparation method of the biological adhesive with high adhesive strength, which comprises the following steps:
A) respectively dissolving super-charge protein and a surfactant in ultrapure water to obtain a super-charge protein solution and a surfactant solution;
the surfactant is a cationic surfactant or an anionic azobenzene surfactant;
the cationic surfactant has a catechol structural unit;
B) dropwise adding the surfactant solution into the super-charge protein solution, and uniformly mixing to obtain an adhesive solution;
C) and centrifuging the adhesive solution, and freeze-drying for 0.2-0.5 h to obtain the biological adhesive with high adhesive strength.
In the present invention, the kind and the amount of the super-charged protein, the cationic surfactant and the anionic azobenzene surfactant are the same as those of the super-charged protein, the cationic surfactant and the anionic azobenzene surfactant, and are not repeated herein.
In the invention, the molar concentration of the super-charged protein solution is preferably 200-300 mu mol/L, and more preferably 220-250 mu mol/L; the molar concentration of the surfactant solution is preferably 10-20 mu mol/L.
After the adhesive solution is obtained, the invention carries out high-speed centrifugation on the adhesive solution, and then the biological adhesive with high adhesive strength is obtained by direct freeze-drying.
In the invention, the speed of centrifugation is preferably 15000-18000 rpm, more preferably 16000-17000 rpm, and most preferably 16800 rpm; the time for centrifugation is preferably 5-20 min, and more preferably 10-15 min.
The freeze-drying time is preferably 0.2-0.5 hour.
The biological adhesive can be applied to the surfaces of steel sheets, aluminum sheets, polyethylene and polyvinyl chloride and shows good adhesion performance.
The invention provides a biological adhesive with high adhesive strength, which consists of super-charged protein and a surfactant; the surfactant is a cationic surfactant or an anionic azobenzene surfactant; the cationic surfactant has a catechol structural unit. The invention utilizes the electrostatic force effect, the pi-cation and the pi-pi accumulation effect between the super charge protein and the surfactant to improve the adhesion performance of the biological protein adhesive. Experimental results show that the biological adhesive disclosed by the invention has excellent adhesive performance on the surfaces of steel sheets, polyethylene and polyvinyl chloride. In addition, the preparation method is simple in preparation process, free of special equipment, low in cost and environment-friendly. Compared with the prior art, the invention has good biocompatibility and degradability.
In order to further illustrate the present invention, the following examples are given to describe the high adhesive strength bioadhesive and the preparation method thereof in detail, but should not be construed as limiting the scope of the present invention.
Example 1
Respectively transforming the super positive charge protein K series carrier plasmids into an escherichia coli expression strain BL21DE3, selecting a monoclonal colony, and carrying out overnight culture by using LB culture solution; adding the overnight activated expression seed liquid into a TB culture medium, adding an inducer isopropyl-beta-D-thiogalactopyranoside when the bacterial liquid reaches an OD600 value of 0.8, and cooling to 30 ℃ for overexpression. Inducing for 12 hours, collecting thalli, resuspending with lysis buffer solution, adding protease inhibitor, DNA enzyme and lysozyme, crushing thalli with a high-pressure crusher, centrifuging at a rotating speed of more than 10000rpm, collecting supernate, and purifying by HPLC to finally obtain the super positive charge protein K. This example shows that the amount of the super-positively charged protein charged is 72.
An unfolded positive charge protein aqueous solution with the concentration of 220 mu mol/L and an azobenzene surfactant aqueous solution with the concentration of 10 mu mol/L are respectively prepared. By mixing the two, shaking 5 minutes after high speed centrifugation at 16800rpm for 10 minutes. And removing the supernatant, and freeze-drying for 0.2 h to obtain the biological adhesive (K72-NAT).
Example 2
A bioadhesive (K108-NAT) was prepared according to the method of example 1, except that the number of charges of the super-positively charged protein in this example was 108.
The bioadhesive of this example was applied to the bottom of two 20mL glass vials, which were adhered together, and subjected to a weighing test, as shown in fig. 2, and as can be seen from fig. 2, the bioadhesive of this example was able to withstand a weight of 600g when applied to a release surface.
Example 3
Respectively transforming the super negative charge protein E series carrier plasmids into an escherichia coli expression strain BL21DE3, selecting a monoclonal colony, and carrying out overnight culture by using LB culture solution; adding the overnight activated expression seed liquid into a TB culture medium, adding an inducer isopropyl-beta-D-thiogalactopyranoside when the bacterial liquid reaches an OD600 value of 0.8, and cooling to 30 ℃ for overexpression. Inducing for 12 hours, collecting thalli, resuspending with lysis buffer solution, adding protease inhibitor, DNA enzyme and lysozyme, crushing thalli with a high-pressure crusher, centrifuging at a rotating speed of more than 10000rpm, collecting supernatant, and purifying by HPLC to finally obtain the super negative charge protein E. The charge per molecule of the E series in this example is 36.
An unfolded negative charge protein aqueous solution with a concentration of 220 mu mol/L and a cationic surfactant aqueous solution with a concentration of 10 mu mol/L are respectively prepared. By mixing the two, shaking 5 minutes after high speed centrifugation at 16800rpm for 10 minutes. The supernatant was discarded and lyophilized for 0.2 hours to obtain a bioadhesive (E36-PDD).
Example 4
A bioadhesive (E72-PDD) was prepared according to the preparation method in example 3, except that the number of charges of the super negative-charged protein in this example was 72.
Example 5
A bioadhesive (E14-PDD) was prepared according to the preparation method in example 3, except that the number of charges of the super negative-charged protein in this example was 144.
The results of the adhesion performance test of the bio-adhesive of examples 1-2 on the surfaces of different materials are shown in fig. 3, and fig. 3 shows the shear strength of the bio-adhesive of examples 1-2 of the present invention on the surfaces of different materials. The adhesive exhibits superior adhesion properties on various substrates, including steel, aluminum, polyethylene, and polyvinyl chloride.
The results of the adhesion performance test of the bio-adhesive in examples 3 to 5 on the surfaces of different materials are shown in fig. 4, and fig. 4 is the shear strength of the bio-adhesive in examples 3 to 5 of the present invention on the surfaces of different materials. The adhesive exhibits good adhesion properties on various substrates including steel, aluminum, polyethylene and polyvinyl chloride.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (4)
1. A biological adhesive with high adhesion strength is prepared from super-positive charge protein and anionic azobenzene surfactant C19H23N2NaO7S;
the super positive charge protein is expressed according to the following steps:
respectively transforming the super positive charge protein K series carrier plasmids into pichia pastoris or escherichia coli expression strains, selecting a monoclonal colony, and carrying out overnight culture by using LB culture solution; adding the overnight activated expression seed liquid into a TB culture medium, adding an inducer isopropyl-beta-D-thiogalactopyranoside when the bacterial liquid reaches an OD600 value of 0.8, and cooling to 30 ℃ for overexpression; inducing for 12 hours, collecting thalli, resuspending with lysis buffer solution, adding protease inhibitor, DNA enzyme and lysozyme, crushing thalli with a high-pressure crusher, centrifuging at a rotating speed of more than 10000rpm, collecting supernate, and purifying by HPLC (high performance liquid chromatography) to finally obtain super positive charge protein K;
the charge number of the super positive charge protein is 72-108;
the molar ratio of the super charge protein to the surfactant is 1: (1-5).
2. The bioadhesive of claim 1, wherein the bioadhesive has a water content of 5% to 15%.
3. The method for preparing a high adhesive strength bioadhesive according to claim 1, comprising the steps of:
A) will be provided withSuper positive charge protein and anionic azobenzene surfactant C19H23N2NaO7S are respectively dissolved in ultrapure water to obtain a super-charge protein solution and a surfactant solution;
B) dropwise adding the surfactant solution into the super-charge protein solution, and uniformly mixing to obtain an adhesive solution;
C) and centrifuging the adhesive solution, and freeze-drying for 0.2-0.5 h to obtain the biological adhesive with high adhesive strength.
4. The method according to claim 3, wherein the centrifugation is performed at a rate of 15000 to 18000 rpm;
the centrifugation time is 5-20 min.
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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 |
| CN114292621B (en) * | 2021-12-29 | 2023-05-02 | 清华大学 | High-low temperature adhesive based on bioengineering protein and preparation method thereof |
| CN117462755A (en) * | 2023-11-24 | 2024-01-30 | 北京康派特医疗器械有限公司 | Biological repair material and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102548581A (en) * | 2009-07-23 | 2012-07-04 | 犹他大学研究基金会 | Adhesive complex coacervates and methods of making and using thereof |
| 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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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004131741A (en) * | 2004-01-20 | 2004-04-30 | Nippi Kollagen Kogyo Kk | Composition for sausage casing, containing adhesive component |
| JP5530934B2 (en) * | 2008-01-24 | 2014-06-25 | ユニバーシティ オブ ユタ リサーチ ファンデーション | Adhesive composite coacervate and methods of making and using the same |
| WO2011025158A2 (en) * | 2009-08-25 | 2011-03-03 | 포항공과대학교 산학협력단 | Coacervate having an ionic polymer mixed with the adhesive protein of a mussel or of a species of the variome thereof |
| US8828542B2 (en) * | 2010-02-26 | 2014-09-09 | Korea University Research And Business Foundation | Nanoparticles |
| KR101677097B1 (en) * | 2015-03-18 | 2016-11-18 | 포항공과대학교 산학협력단 | Adhesive sealant for treating of fistula, perforation or anastomosis leak and for connecting of internal organ |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102548581A (en) * | 2009-07-23 | 2012-07-04 | 犹他大学研究基金会 | Adhesive complex coacervates and methods of making and using thereof |
| 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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