CN105734039A - Preparation method of polydopamine/graphene oxide composite film with muramidase immobilized - Google Patents
Preparation method of polydopamine/graphene oxide composite film with muramidase immobilized Download PDFInfo
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Abstract
The invention discloses a preparation method of a polydopamine/graphene oxide composite film with muramidase immobilized.The method includes the steps that firstly, autopolymerization of dopamine is carried out on the surface of a substrate like glass, an incomplete graphene oxide film layer is introduced onto a polydopamine film layer, part of the zone of the polydopamine film layer is covered with graphene oxide by controlling the concentration of graphene oxide and the temperature of the soaking process, the oxygen-containing functional group part of graphene oxide is reduced by polydopamine at the same time in the process, and a partially-reduced graphene oxide film layer is formed; finally, muramidase is immobilized to the incomplete partially-reduced graphene oxide film layer, and the polydopamine/graphene oxide composite film with muramidase immobilized is obtained.The method is simple and easy to implement; by inserting the incomplete graphene oxide film layer, the enzyme loading capacity of the composite film is obviously increased, the antimicrobial property is remarkably improved, and long-term high-efficiency bacteriostasis can be achieved; the composite film has good application prospects in the fields of food safety, medical treatment and public health and the like.
Description
Technical field
The present invention relates to the preparation method field of graphene oxide composite material, the preparation method particularly relating to a kind of poly-dopamine/graphene oxide composite membrane with high anti-microbial property and load enzyme amount.
Background technology
Antibacterial glass or packaging bag are as a kind of novel ecological functional material, it is possible to be widely used in the fields such as medical treatment, food safety and public health.Now common antibiotic glass adopts and adds ionized metal such as zinc, silver-bearing copper etc. in the manufacture process of glass, or at glass surface titanium dioxide coating film, plays antibacterial action by the photocatalysis of titanium dioxide.But human body is still had certain harm by these antibacterial, and is discharged in environment and can cause secondary pollution.Therefore, need Environment badly more friendly, harmless anti-fouling agent.
At present, studying more is select protein-based organic antibacterial agent, wherein, has been reported that and shows, by poly-dopamine rete load lysostaphin, it is possible to make matrix show comparatively excellent anti-microbial property.But still there is some problems, the activity such as protein is shorter, less stable, thus leverages it and use.Therefore, how to improve the load capacity of protein-based antibacterial, and extend its antibacterial time and just become one of study hotspot.
Summary of the invention
For the problems referred to above, it is an object of the invention to provide a kind of load capacity high, the preparation method of the poly-dopamine/graphene oxide composite membrane for fixing lysozyme of antibacterial time length.
First the present invention carries out the auto polymerization of dopamine at matrix surfaces such as glass, thus being coated with poly-dopamine rete one layer complete at matrix surface；Incomplete graphene oxide rete is introduced again on poly-dopamine rete, by controlling the concentration of graphene oxide, regulate the temperature in immersion process, the subregion making poly-dopamine rete is covered with graphene oxide, outside the poly-dopamine rete in remainder region is still exposed to, in the process, the oxygen-containing functional group part of graphene oxide is gathered dopamine reduction, forms part redox graphene rete simultaneously；Finally on incomplete partial reduction graphene oxide rete, fix lysozyme, prepare the poly-dopamine/graphene oxide composite membrane of fixing lysozyme.
Graphene oxide of the present invention is the graphene oxide that hummers method is prepared, owing to it has bigger reference area, more oxygen-containing functional group, lysozyme is had good fixation, and itself there is good anti-microbial property, can fixed oxygen functionalized graphene preferably by the modified matrix of poly-dopamine.The poly-dopamine rete formed after polymerization in the present invention its there is good adhesion property, therefore lysozyme and graphene oxide can be carried out good fixing.The matrix of the poly-dopamine/partial reduction graphene oxide rete of cladding can significantly promote lysozyme fixed amount, extends its antibacterial time simultaneously.
The present invention comprises the steps:
(1) glass surface carries out dopamine modifiy
Dopamine or its salt being joined and be configured to dopamine solution in alkaline solution, put into cleaned matrix, room temperature stands 24 hours, and ultrasonic cleaning after taking-up, nitrogen dries up, to be coated with one or more layers complete poly-dopamine rete at matrix surface；
(2) preparation of imperfect Graphene derivative rete
The matrix of poly-for cladding dopamine rete is put in the graphene oxide solution of 0.1～0.5mg/mL, soak 3 hours under 25～80 DEG C of conditions, forming portion segregation dopamine rete is coated with the incomplete graphene oxide rete of graphene oxide, ultrasonic cleaning after taking-up, nitrogen dries up, owing to graphene oxide part is gathered dopamine reduction, on poly-dopamine rete, finally form incomplete partial reduction graphene oxide rete；
(3) antiseptic protein is fixing
The PBS solution of pH7.4 will add antiseptic protein, stir half an hour under 0～4 DEG C of condition, put into the matrix of the poly-dopamine of cladding and incomplete partial reduction graphene oxide rete, soak 48 hours under 4 DEG C of conditions, rinse with PBS after taking-up, be fixed the poly-dopamine/graphene oxide composite membrane of lysozyme.
Described alkaline solution is Tris solution or the alkaline aqueous solution (such as KOH, NaOH) of pH6.5～8.5, and concentration is 0.1～0.5mg/mL.
Described antiseptic protein is the protein that animal lysozyme, microbic muramidase or lysostaphin etc. have antibiotic and sterilizing performance.
The described preferred hen's egg-white lysozyme of animal lysozyme.
Described basic material is glass, metal or plastics.
Compared with prior art, present invention have the advantage that
(1) poly-dopamine rete and incomplete graphene oxide rete are combined by the present invention, the poly-dopamine/graphene oxide composite film of preparation, or have certain antibacterial action by the composite membrane itself of preparation after layer assembly.
(2) present invention adopts Hen egg-white lysozyme as one of main antibacterial, and it has good anti-microbial property, simultaneously harmless, is eco-friendly nontoxic antibacterial.
(3) present invention is by inserting incomplete graphene oxide rete, makes composite membrane load enzyme amount significantly improve, and anti-microbial property is obviously improved, simultaneously more existing similar antibacterial, and antibacterial time is also significantly increased.
Accompanying drawing explanation
Fig. 1 is graphene oxide (GO) (a), through poly-dopamine/redox graphene (PDA/rGO) composite membrane modified glass surface (b) with through gathering the infrared spectrum of the modified glass surface (c) of dopamine/redox graphene-lysozyme (PDA/rGO-cLY) composite membrane.
Fig. 2 is the glass surface (a) that PDA/rGO is modified respectively, the partial enlargement (b) of the glass surface that PDA/rGO is modified；The stereoscan photograph of the glass surface (c) that PDA/rGO-cLY is modified.
Fig. 3 is escherichia coli (b) stereoscan photograph of escherichia coli (a) lethal after lysozyme effect and oxidized Graphene acting lethality.
Fig. 4 is growth tendency comparison diagram (a) with blank elements, 7 hours interior growth tendency contrast enlarged drawing (b) in escherichia coli 18 hours containing the glass modified through the PDA-cLY glass modified and PDA/rGO-cLY in culture fluid.
Fig. 5 be through PDA-cLY modify glass and through PDA/rGO-cLY modify glass at different time points sterilizing rate comparison diagram.
Fig. 6 is through glass surface (a) modified for PDA-cLY with through the fluorescent microscopy images of glass surface (b) modified for PDA/rGO-cLY.
Fig. 7 is PDA/rGO stereoscan photograph after 0.1mg/mL graphene oxide solution (a), 0.3mg/mL graphene oxide solution (b) and 0.5mg/mL graphene oxide solution (c) soak 3 hours under 50 DEG C of conditions respectively.
Fig. 8 is graphene oxide solution when being 0.3mg/mL, respectively at 25 DEG C (a), 50 DEG C (b) and 80 DEG C soak 3 hours after PDA/rGO stereoscan photograph.
Detailed description of the invention
Below in conjunction with accompanying drawing and comparative example, embodiments of the invention are described in detail, so that advantages and features of the invention can be easier to be readily appreciated by one skilled in the art, thus protection scope of the present invention being made apparent clear and definite defining.
Comparative example 1: glass surface is carried out PDA-cLY rete modification.
Reaction raw materials is the Tris solution of dopamine hydrochloride, pH8.5, Hen egg-white lysozyme, the PBS of pH7.4.Wherein dopamine hydrochloride is purchased from Sigma-Aldrich, and Hen egg-white lysozyme is purchased from Sigma-Aldrich, and active unit is～100000U/mg, and molecular weight is the strand of 14.3kDa.
(1) glass surface carries out dopamine modifiy
Being joined by dopamine in the Tris solution of pH8.5 and be configured to 2mg/mL, put into cleaned microscope slide, room temperature stands 24 hours, and ultrasonic cleaning after taking-up, nitrogen dries up.
(2) antiseptic protein is fixing
By the PBS solution of pH7.4 adds Hen egg-white lysozyme anti-microbial type protein, stir half an hour under 0～4 DEG C of condition, put into the sheet glass processed through dopamine, soak 48 hours under 4 DEG C of conditions, rinse with PBS after taking-up.
The antibacterial test of prepared antibiotic glass is as follows:
(1) preparation of bacteria suspension
By escherichia coli (E.coli) strain be linked in LB fluid medium and activate, bacterium solution is shaken 24 hours under 37 DEG C of conditions in isothermal vibration incubator.Then pipetting the 6 μ L strains activated so that it is be dispersed in the solution of 8mLLB fluid medium and PBS 1:1, carry out concussion and shake up, put into treated antibiotic glass, concussion cultivation under 37 DEG C of conditions in thinking is cultivated in earthquake.
(2) qualitative antibacterial test
After being taken out by sheet glass after cultivation a period of time, by bacteria adhension, utilize scanning electron microscope that it is observed.
(3) quantitative antibacterial test
To cultivate 1 hour, 3 hours, 5 hours, 7 hours, 13 hours and bacterium solution on 18 hours each time points are taken out, respectively with, after normal saline dilution, utilizing plate count to carry out observing statistics, and utilize sterilizing rate formula that data are added up.
Embodiment 1:PDA/rGO-cLY rete modification is carried out for matrix with glass.
Reaction raw materials is the Tirs solution of dopamine hydrochloride, pH8.5, graphene oxide, Hen egg-white lysozyme, the PBS of pH7.4.Wherein dopamine hydrochloride is purchased from Sigma-Aldrich, and Hen egg-white lysozyme is purchased from Sigma-Aldrich, and active unit is～100000U/mg, and molecular weight is the strand of 14.3kDa, graphene oxide Hummers method (Marcano, D.C.;Kosynkin,D.V.;Berlin,J.M.;Sinitskii,A.;Sun,Z.;Slesarve,A.;Alemany,L.B.;Lu,W.;Tour, J.M.ImprovedSynthesisofGrapheneOxide.ACSNano2010,4,4806-4814.) prepare.
(1) glass surface carries out dopamine modifiy
Being joined by dopamine in the Tris solution of pH8.5 and be configured to 2mg/mL, put into cleaned microscope slide, room temperature stands 24 hours, and ultrasonic cleaning after taking-up, nitrogen dries up.
(2) preparation of imperfect Graphene derivative rete
The Tris solution of pH8.5 is joined in graphene oxide, is configured to 0.3mg/mL graphene oxide solution, adjust pH to 8.5, the sheet glass processed through dopamine is put in graphene oxide solution, soaking 3 hours under 50 DEG C of conditions, ultrasonic cleaning after taking-up, nitrogen dries up.
(3) antiseptic protein is fixing
By the PBS solution of pH7.4 adds Hen egg-white lysozyme, stir half an hour under 0～4 DEG C of condition, put into the sheet glass processed through dopamine and graphene oxide, soak 48 hours under 4 DEG C of conditions, rinse with PBS after taking-up.
Prepared material is by infrared test, such as Fig. 1, compared with simple graphene oxide, is fixed on the imperfect graphene oxide rete in the substrate of glass that poly-dopamine is modified at 1731cm-1(C=O key) and 1402cm-1(C-OH key) part changes.The two peak almost disappears on PDA/rGO spectral line, it follows that partial oxidation of graphite alkene is gathered dopamine reduction, it is determined that end product is PDA/rGO.For the glass infrared spectrum modified through PDA/rGO-cLY, belong to the 3100cm of N-H stretching vibration-1And 2944cm-1Peak, position almost disappears, it was shown that Hen egg-white lysozyme has part to occur and chemical bonds with PDA/rGO.Prove that the oxygen-containing functional group that graphene oxide enriches can provide avtive spot and lysozyme generation chemical action, thus lysozyme is firmly fixed to film surface.
Prepared material is by stereoscan photograph, and such as Fig. 2, it can be seen that for PDA/rGO rete, graphene oxide rete covers above poly-dopamine rete, and rete imperfect, in being partially covered on poly-dopamine film surface.Graphene oxide rete is sprawled more uniform, and there is partial folds centre.For PDA/rGO-cLY rete, Hen egg-white lysozyme is mainly distributed on the marginal portion of fold part and graphene oxide, also has a small amount of distribution on exposed poly-dopamine rete.Prove that bigger serface and the abundant oxygen-containing functional group of graphene oxide can effectively fix lysozyme.
The antibacterial test of prepared antibiotic glass is as follows:
(1) preparation of bacteria suspension
By escherichia coli (E.coli) strain be linked in LB fluid medium and activate, bacterium solution is shaken 24 hours under 37 DEG C of conditions in isothermal vibration incubator.Then pipetting the 6 μ L strains activated so that it is be dispersed in the solution of 8mLLB fluid medium and PBS 1:1, carry out concussion and shake up, put into treated antibiotic glass, concussion cultivation under 37 DEG C of conditions in thinking is cultivated in earthquake.
(2) qualitative antibacterial test
After being taken out by sheet glass after cultivation a period of time, by bacteria adhension, utilize scanning electron microscope that it is observed, such as Fig. 3.Owing to the sterilization mechanism of lysozyme is the Peptidoglycan acting on cell wall, lysozyme embodies its bacteriostasis by the main component of dissolved cell wall.Schemed it can be seen that antibacterial is completely dissolved by a, only surplus residue.And graphene oxide is mainly through the machine cuts effect of physics, cell is blocked at sharp edge, b scheme it can be seen that one section of antibacterial is directly cut off.Therefore, can be seen that, this PDA/rGO composite film itself has certain anti-microbial property, and after lysozyme is fixing, the not edge of passive oxidation Graphene, the antibacterial action of graphene oxide yet suffers from, and graphene oxide is also without causing lysozyme to inactivate, the bactericidal action that lysozyme can also play.
(3) quantitative antibacterial test
To cultivate 1 hour, 3 hours, 5 hours, 7 hours, 13 hours and bacterium solution on 18 hours each time points are taken out, respectively with, after normal saline dilution, utilizing plate count to carry out observing statistics, and utilize sterilizing rate formula that data are added up.
Such as Fig. 4.By scheming a it can be seen that in 18 hours incubation times, best through the PDA/rGO-cLY glass bacteriostasis property modified, in 7 hours cultivated, bacterial reproduction is obvious hardly, in 7 to 13 hours, has obvious bacterial reproduction sign, bacteria growth is slower, have obvious rising after 13 hours, but to 18 little constantly, still have higher sterilizing rate, in conjunction with Fig. 5,18 little sterilizing rates constantly still reach more than 90%.And after cultivating 7 hours, there is significant change through the PDA-cLY glass modified, bacteria growth is significantly raised, and in conjunction with Fig. 5, sterilizing rate is less than 60%.For 7 hours interior bacterial growth trend of clearer observation, be can be seen that by figure b, for the glass that the PDA-cLY glass modified and PDA/rGO-cLY are modified, two components there occurs the phenomenon that obvious sterilizing ability declines all after 5h, but PDA-cLY component becomes apparent from, in conjunction with Fig. 5, it is also possible to obtain corresponding conclusion.It is hereby achieved that, after adding incomplete graphene oxide rete, it is possible to extend the antibacterial time that material is antibacterial.
Comparative example 2:Glass surface is carried out PDA-cLY rete modification.
Reaction raw materials is the Tirs solution of dopamine hydrochloride, pH8.5, Hen egg-white lysozyme, the PBS of pH7.4.Wherein dopamine hydrochloride is purchased from Sigma-Aldrich, and Hen egg-white lysozyme is purchased from Sigma-Aldrich, and active unit is～100000U/mg, and molecular weight is the strand of 14.3kDa.Isothiocyanic acid (FITC) fluorescein is purchased from Sigma-Aldrich.
Lysozyme isothiocyanic acid (FITC) fluorescein is carried out labelling.Configuration pH is carbonate and the sodium chloride bout solution 100mL of 9, adds 1g lysozyme, and dispersing and dissolving is also cooled to 4 DEG C.In 4 DEG C of lysozyme solns, add 10mgFITC, lucifuge magnetic agitation 12 hours in ice-water bath, generate fluorescent marker protein FITC-cLY.Configure excessive semi-saturation ammonium sulfate, join in above-mentioned product, by labelled protein precipitate and separate, go out unconjugated fluorescein.Configuration pH is the phosphate buffer of 7.2, labelled protein of dialysing at 4 DEG C, goes out unnecessary ammonium sulfate, to can't detect ammonium ion and fluorescein in the solution dialysed.After product is carried out lyophilization, keep in Dark Place under 4 DEG C of conditions, stand-by.
(1) glass surface carries out dopamine modifiy
Being joined by dopamine in the Tris solution of pH8.5 and be configured to 2mg/mL, put into cleaned microscope slide, room temperature stands 24 hours, and ultrasonic cleaning after taking-up, nitrogen dries up.
(2) antiseptic protein is fixing
By the PBS solution of pH7.4 adds labeled after Hen egg-white lysozyme, under 0～4 DEG C of condition stir half an hour, put into the sheet glass processed through dopamine, under 4 DEG C of conditions soak 48 hours, after taking-up with PBS rinse, stand-by.
Utilize fluorescence microscope, when excitation wavelength is 490nm, its load enzyme amount is observed.
Embodiment 2:Glass surface is carried out PDA/rGO-cLY rete modification.
Reaction raw materials is the Tirs solution of dopamine hydrochloride, pH8.5, Hen egg-white lysozyme, the PBS of pH7.4.Wherein dopamine hydrochloride is purchased from Sigma-Aldrich, and Hen egg-white lysozyme is purchased from Sigma-Aldrich, and active unit is～100000U/mg, and molecular weight is the strand of 14.3kDa.Prepared by graphene oxide Hummers method.Isothiocyanic acid (FITC) fluorescein is purchased from Sigma-Aldrich.
Lysozyme isothiocyanic acid (FITC) fluorescein is carried out labelling.Configuration pH is carbonate and the sodium chloride bout solution 100mL of 9, adds 1g lysozyme, and dispersing and dissolving is also cooled to 4 DEG C.In 4 DEG C of lysozyme solns, add 10mgFITC, lucifuge magnetic agitation 12 hours in ice-water bath, generate fluorescent marker protein FITC-cLY.Configure excessive semi-saturation ammonium sulfate, join in above-mentioned product, by labelled protein precipitate and separate, go out unconjugated fluorescein.Configuration pH is the phosphate buffer of 7.2, labelled protein of dialysing at 4 DEG C, goes out unnecessary ammonium sulfate, to can't detect ammonium ion and fluorescein in the solution dialysed.After product is carried out lyophilization, keep in Dark Place under 4 DEG C of conditions, stand-by.
(1) glass surface carries out dopamine modifiy
Being joined by dopamine in the Tris solution of pH8.5 and be configured to 2mg/mL, put into cleaned microscope slide, room temperature stands 24 hours, and ultrasonic cleaning after taking-up, nitrogen dries up.
(2) preparation of imperfect Graphene derivative rete
The Tris solution of pH8.5 is joined in graphite oxide, is configured to the graphene oxide solution of 0.3mg/mL, adjust pH to 8.5, the sheet glass processed through dopamine is put in graphene oxide solution, soak 3 hours under 50 DEG C of conditions, going out rear ultrasonic cleaning, nitrogen dries up.
(3) antiseptic protein is fixing
The PBS solution of pH7.4 will add labeled Hen egg-white lysozyme, stir half an hour under 0～4 DEG C of condition, put into the sheet glass processed through dopamine and Graphene derivative, soak 48 hours under 4 DEG C of conditions, rinse with PBS after taking-up, stand-by.
Utilize fluorescence microscope, when excitation wavelength is 490nm, its load enzyme amount is observed.The enzyme of labelling is shown as yellow-green fluorescence, such as Fig. 6.As seen from the figure, will become apparent from the fluorescent labeling containing two kinds of forms in figure b, wherein brighter is the lysozyme fixed through electrostatic interaction, and dark part shows as the lysozyme fixing by covalent bond effect.For figure a it can be seen that for PDA/rGO composite membrane, its immobilized form is more, and the amount of load enzyme is bigger.Prove that the imperfect rete of its rGO plays a very important role in composite film.
Embodiment 3-5:When variable concentrations, glass surface is carried out the preparation of PDA/rGO rete.
Reaction raw materials is the Tirs solution of dopamine hydrochloride, pH8.5, graphene oxide.Wherein dopamine hydrochloride is purchased from Sigma-Aldrich, graphene oxide Hummers method.
(1) glass surface carries out dopamine modifiy
Being joined by dopamine in the Tris solution of pH8.5 and be configured to 2mg/mL, put into cleaned microscope slide, room temperature stands 24 hours, and ultrasonic cleaning after taking-up, nitrogen dries up.
(2) preparation of imperfect Graphene derivative rete
The Tris solution of pH8.5 is joined in graphene oxide, it is configured to 0.1mg/mL, 0.3mg/mL and 0.5mg/mL graphene oxide solution respectively, adjust pH to 8.5, the sheet glass processed through dopamine is put in graphene oxide solution, soak 3 hours under 50 DEG C of conditions, ultrasonic cleaning after taking-up, nitrogen dries up.
Utilize scanning electron microscope that its distribution situation is observed, such as Fig. 7.Can be seen that in 0.1mg/mL, 0.3mg/mL and the 0.5mg/mL graphene oxide solution of pH8.5, soak 3 sheet glass as a child under 50 DEG C of conditions and be respectively formed incomplete graphene oxide rete, and along with the increase of concentration, wrinkle have the trend increased.
(3) antiseptic protein is fixing
By the PBS solution of pH7.4 adds Hen egg-white lysozyme, stir half an hour under 0～4 DEG C of condition, put into the sheet glass processed through dopamine and Graphene derivative, soak 48 hours under 4 DEG C of conditions, rinse with PBS after taking-up, stand-by.
Embodiment 6-8: under condition of different temperatures, glass surface is carried out the preparation of PDA/rGO rete.
Reaction raw materials is the Tirs solution of dopamine hydrochloride, pH8.5, graphene oxide.Wherein dopamine hydrochloride is purchased from Sigma-Aldrich, graphene oxide Hummers method.
(1) glass surface carries out dopamine modifiy
Being joined by dopamine in the Tris solution of pH8.5 and be configured to 2mg/mL, put into cleaned microscope slide, room temperature stands 24 hours, and ultrasonic cleaning after taking-up, nitrogen dries up.
(2) preparation of imperfect Graphene derivative rete
The Tris solution of pH8.5 is joined in graphene oxide, it is configured to 0.3mg/mL graphene oxide solution, adjust pH to 8.5, the sheet glass processed through dopamine is put in graphene oxide solution, soak 3 hours under 25 DEG C, 50 DEG C and 80 DEG C of conditions respectively, ultrasonic cleaning after taking-up, nitrogen dries up.
Utilize scanning electron microscope that its distribution situation is observed, such as Fig. 8.Can be seen that the sheet glass after soaking 3 hours under 25 DEG C, 50 DEG C and 80 DEG C of temperature conditions is respectively formed incomplete graphene oxide rete, and along with the rising of temperature, graphene oxide is combined with, with poly-dopamine rete, the trend becoming big.
(3) antiseptic protein is fixing
By the PBS solution of pH7.4 adds Hen egg-white lysozyme, stir half an hour under 0～4 DEG C of condition, put into the sheet glass processed through dopamine and Graphene derivative, soak 48 hours under 4 DEG C of conditions, rinse with PBS after taking-up, stand-by.
Claims (6)
1. the preparation method of the poly-dopamine/graphene oxide composite membrane of an immobilized lysozyme, it is characterised in that comprise the following steps:
(1) dopamine or dopamine hydrochloride being joined and be configured to dopamine solution in alkaline solution, put into cleaned matrix, room temperature stands 24 hours, and ultrasonic cleaning after taking-up, nitrogen dries up, to be coated with one or more layers complete poly-dopamine rete at matrix surface；
(2) matrix of poly-for cladding dopamine rete is put in graphene oxide solution, soak 3 hours under 25 DEG C～80 DEG C conditions, forming portion segregation dopamine rete is coated with the incomplete graphene oxide rete of graphene oxide, ultrasonic cleaning after taking-up, nitrogen dries up, owing to graphene oxide part is gathered dopamine reduction, on poly-dopamine rete, finally form incomplete partial reduction graphene oxide rete；
(3) PBS solution of pH7.4 will add antiseptic protein, stir half an hour under 0～4 DEG C of condition, put into the matrix of the poly-dopamine of cladding and incomplete partial reduction graphene oxide rete, soak 48 hours under 4 DEG C of conditions, rinse with PBS after taking-up, be fixed the poly-dopamine/graphene oxide composite membrane of lysozyme.
2. preparation method according to claim 1, it is characterised in that described alkaline solution is Tris solution or the alkaline aqueous solution of pH6.5～8.5.
3. preparation method according to claim 1, it is characterised in that the concentration of described graphene oxide solution is 0.1～0.5mg/mL.
4. preparation method according to claim 1, it is characterised in that described antiseptic protein is animal lysozyme, microbic muramidase or lysostaphin.
5. preparation method according to claim 4, it is characterised in that the described preferred hen's egg-white lysozyme of animal lysozyme.
6. preparation method according to claim 1, it is characterised in that described basic material is glass, metal or plastics.
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| CN111529682A (en) * | 2020-05-11 | 2020-08-14 | 中国人民解放军陆军军医大学第一附属医院 | Chemotaxis antibacterial nano material and preparation method and application thereof |
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| CN108911857A (en) * | 2018-08-09 | 2018-11-30 | 长沙昱旻信息科技有限公司 | A kind of multifunctional slow-release organic fungi-manure |
| CN109576257A (en) * | 2018-12-04 | 2019-04-05 | 清华大学 | A kind of enzyme catalyst and preparation method thereof of part photo-thermal effect |
| CN111529682A (en) * | 2020-05-11 | 2020-08-14 | 中国人民解放军陆军军医大学第一附属医院 | Chemotaxis antibacterial nano material and preparation method and application thereof |
| CN111744051A (en) * | 2020-07-15 | 2020-10-09 | 中国人民解放军西部战区总医院 | Preparation method and wound healing method of graphene oxide-lysozyme/alkaline fibroblast growth factor composite dressing |
| CN113414497A (en) * | 2021-07-16 | 2021-09-21 | 北京理工大学重庆创新中心 | Method for processing and preparing surface micro-nano composite structure |
| CN113414497B (en) * | 2021-07-16 | 2022-07-29 | 北京理工大学重庆创新中心 | Method for processing and preparing surface micro-nano composite structure |
| CN114933419A (en) * | 2022-04-29 | 2022-08-23 | 北京仿生界面科学未来技术研究院 | Method for preparing ultrathin flexible glass by utilizing bionic weathering enzyme composite glass thinning agent |
| CN114933419B (en) * | 2022-04-29 | 2023-09-29 | 北京仿生界面科学未来技术研究院 | Method for preparing ultrathin flexible glass by using bionic weathering enzyme composite glass thinning agent |
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