CN114100676A - A kind of synthetic method and application of functionalized modified metal nanoparticles - Google Patents
A kind of synthetic method and application of functionalized modified metal nanoparticles Download PDFInfo
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- C—CHEMISTRY; METALLURGY
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Abstract
本发明公开了一种功能化修饰金属纳米颗粒的合成方法与应用。所述合成方法包括：将耐辐射奇球菌压力破碎后取上清液，通过盐析法制备蛋白提取物作为合成剂；向所述合成剂中加入Ag+盐溶液反应后，再加入Au3+盐溶液反应，合成蛋白修饰的金银双金属纳米颗粒。制得的功能化修饰金属纳米颗粒可用于降解活性黑及番红O染料。The invention discloses a synthesis method and application of functionalized modified metal nanoparticles. The synthesis method comprises: crushing Deinococcus radiodurans under pressure and taking the supernatant, preparing a protein extract as a synthesis agent by a salting-out method; adding Ag + salt solution to the synthesis agent for reaction, and then adding Au 3+ Salt solution reaction to synthesize protein-modified gold-silver bimetallic nanoparticles. The prepared functionalized modified metal nanoparticles can be used to degrade reactive black and safranin O dyes.
Description
Technical Field
The invention belongs to the crossing field of nano materials and bioengineering, and particularly relates to a synthesis method and application of functionalized modified metal nanoparticles.
Background
The bulk noble metal has good ductility, electrical conductivity, and thermal conductivity, and when the size is small to a specific nanometer level, the bulk noble metal can show unique surface effect and physicochemical property. Because the metal nano particles have small size and good stability, and have special photoelectric characteristics and biological safety, the metal nano particles have wide application value in the biomedical fields of bacteriostat, gene transportation, enzyme carriers, biological imaging and the like.
Bimetallic nanoparticles (Bimetallic nanoparticles) are nanoparticles formed by combining two different metals in various combinations, such as alloys, shell-core structures or mutual contact. Bimetallic nanoparticles generally have a larger specific surface area and a lower density than monometallic nanoparticles. Each of which has different properties, and due to the synergistic effect, bimetallic nanoparticles tend to exhibit novel electronic, optical and catalytic properties, with more variation in properties resulting from adjustment of the ratio of metals therein.
At present, the preparation of bimetallic nanoparticles can be divided into physical, chemical and biological methods according to different experimental methods. Vargas-Hernandez et al reported the physical process of microwave heating for the rapid formation of nanoparticles of Au-Ag core-shell structure in an article entitled "A Synthesis route of gold nanoparticles with out using a reduction agent" (Applied Physics Letters,2010,96(21): 145). There are various chemical preparation methods including co-chemical reduction, photochemical reduction, displacement reaction, electrochemical method, etc., such as Kumar et al, in the article entitled "Synthesis and characterization of SAu interaction in gold Nanoparticle bound polymeric beads" (Journal of Nanoparticle Research,2004,6(4): 369-. The conventional physical and chemical methods have the defects of serious pollution, residual product surface, higher cost and the like.
Compared with the prior art, the biological synthesis method is more environment-friendly and has the characteristic of good biocompatibility. The biological method is simple and rapid, has low toxic and side effects, is green and environment-friendly, and can synthesize products with definite sizes and forms under the premise of optimizing conditions. Biological synthesis of metal nanoparticles various biological systems can be utilized, including bacteria, fungi, plant extracts and their biomolecules, among others.
Synthesis of plants: plant cells contain many active ingredients, and various plant materials can be used as reducing agents and stabilizing agents required for nanoparticle synthesis. For example, the water chestnut skin extract can mediate the synthesis of gold-silver alloy nanoparticles.
Microbial synthesis: a variety of microbial cells may be involved in the synthesis of metal nanoparticles. The microorganism is generally tenacious in vitality and can resist the toxic pressure brought by metal ions; the product has various metabolic pathways and active products, and has complexing and depositing effects on metals; the propagation passage time is short, thereby being suitable for large-scale production. The reaction condition of the microbial synthesis method is mild, the yield can be increased or the physical characteristics of the product can be changed by modification, and the method is very suitable for synthesizing nano materials as a substrate. Metal nanoparticles can be synthesized biologically by various microorganisms, such as Stenotrophomonas maltophilia, Zygomycete, Bacillus brevis [59], Shewanella, and the like.
Biomolecule synthesis: many biological macromolecules have extremely strong reducing power. When microbial cells are used as substrates, the ligand components on the surface of the metal particles are complex due to various metabolites, and the physicochemical properties of the nanoparticles are not uniform; the active biological molecules with higher purity are used as substrates, so that the particle size and the form of the product can be more controllable, and DNA, nucleic acid, amino acid, polypeptide, protein, pigment, cellulose and the like can be directly used for synthesis. The reaction nature of the synthesis by using the biological molecules is the same as that of a microbial cell method, but the synthesized nano particles have smaller size and more single product, and have more excellent performance. Meanwhile, the method retains the advantage of high biocompatibility of a biological cell synthesis product, and is safer and more controllable than a chemical method.
Disclosure of Invention
In view of the above, the invention provides a synthesis method and application of functionalized modified metal nanoparticles, which are used for synthesizing gold and silver bimetallic nanoparticles by a deinococcus radiodurans protein extract biological method and effectively degrading active black dyes and safranin O dyes.
A synthetic method of functionalized modified metal nanoparticles comprises the following steps:
(1) crushing deinococcus radiodurans under pressure, collecting supernatant, and preparing protein extract by salting out method as synthesis agent;
(2) adding Ag to the synthesis agent+After the salt solution reacts, Au is added3+And (3) salt solution reaction to synthesize the protein-modified gold-silver bimetallic nano-particles.
In one embodiment of the present invention, a method for synthesizing functionalized metal nanoparticles comprises the following steps:
(1) resuspending deinococcus radiodurans in logarithmic phase with phosphate buffer solution, sufficiently oscillating, performing cell disruption at 4 deg.C under 1200bar for 3min, centrifuging the disruption solution at 15,000 Xg for 30min, and collecting supernatant; adding ammonium sulfate into the supernatant according to the proportion of 80g of ammonium sulfate to every 100mL of supernatant, standing for 4 hours at 4 ℃, centrifuging for 10min at 12,000 Xg, and collecting the precipitate obtained by centrifugation; dissolving the precipitate with ultrapure water, dialyzing with a cellulose acetate dialysis membrane with a cut-off of 14kDa for over 24 hours, and concentrating the dialyzed solution to a protein concentration of 2mg/ml to obtain a protein extract solution as a synthesis agent;
(2) adding Ag to the synthesis agent+Salt solution to Ag+Reacting at a final concentration of 1mM at 37 deg.C and pH 7.0 for 12 hr or more, centrifuging at 20,000 Xg for 10min, and collecting the precipitate; washing the precipitate with ultrapure water, centrifuging for 10min at 12,000 Xg, repeating washing and centrifuging for several times, re-suspending the precipitate with ultrapure water, dialyzing with 14kDa cellulose acetate dialysis membrane for over 24 hr, centrifuging for 10min at 20,000 Xg, dissolving the precipitate in ultrapure water, adding Au, and centrifuging3+The salt solution was mixed well until Au3+The final concentration of the gold and silver nanoparticles is 1mM, and the reaction is carried out for more than 2 hours under the conditions of pH 7.0 and 37 ℃ to obtain a reaction solution containing the protein-modified gold and silver bimetallic nanoparticles.
In an embodiment of the present invention, the synthesis method further comprises: and purifying the reaction solution to obtain the gold and silver bimetallic nanoparticles modified by the protein.
In one embodiment of the present invention, the synthesis method comprises the following steps: filtering the reaction solution by using a 0.22 mu m needle filter, centrifuging 20,000 Xg of obtained filtrate for 10min, washing the obtained precipitate by centrifugation with ultrapure water, centrifuging for 10min again by using 20,000 Xg, repeatedly washing and centrifuging for a plurality of times, dissolving the precipitate by using the ultrapure water, centrifuging for more than 24 hours by using a cellulose acetate dialysis membrane with the interception amount of 14kDa, and freeze-drying the solution after dialysis to obtain the protein-modified gold and silver bimetallic nanoparticles.
In one embodiment of the present invention, in step (1), the concentration is performed by centrifugation using an ultrafiltration concentration tube of 4,800 × g.
In one embodiment of the present invention, in step (2), Ag+The salt solution is silver nitrate solution, Au3+The salt solution is chloroauric acid (HAuCl)4·3H2O) solution.
In one embodiment of the present invention, step (2) is performed with Ag+The salt solution was reacted for 12 hours with Au3+The salt solution reaction time was 6 hours.
The invention also provides the protein-modified gold-silver bimetallic nano-particles prepared by the synthesis method.
The gold and silver bimetallic nanoparticles modified by the protein obtained by the synthesis method have characteristic absorption peaks around 550nm through UV/Vis wavelength detection; by EDS analysis, there is a characteristic peak for Au at the 2.1keV position, a characteristic peak for Ag at the 3.0keV position, and also proteins; the average hydrated particle size was 149.8nm as determined by DLS; the crystal structure has characteristic crystal faces of Ag and Au through XRD detection.
In addition, the invention also provides an application of the functionalized modified metal nanoparticle, which comprises the following steps: the gold and silver bimetallic nanoparticles modified by the protein are used for degrading active black dye.
In addition, the invention also provides an application of the functionalized modified metal nanoparticle, which comprises the following steps: the protein-modified gold and silver bimetallic nanoparticles are used for degrading safranin O dye.
Compared with the prior art, the invention has the following beneficial technical effects:
the invention uses Deinococcus radiodurans protein extract to synthesize protein-modified gold and silver bimetallic nanoparticles, the product has unique nano-size and bimetallic characteristic crystal face, meanwhile, because the synthetic raw material is from Deinococcus radiodurans (Deinococcus radiodurans) with strong viability and high propagation efficiency, the protein extract is rich in oxidation resistance related protein, and key substances are provided for preparing the functionally-modified gold and silver bimetallic nanoparticles. The protein-modified gold and silver bimetallic nano-particles synthesized by the method can be used for efficiently degrading active black dye and safranin O dye.
Drawings
Fig. 1 is an ultraviolet-visible light absorption spectrum (UV/Vis) spectrum of the gold-silver bimetallic nanoparticle prepared in example 1 of the present invention.
Fig. 2 is a Scanning Electron Microscope (SEM) image of the gold and silver bimetallic nanoparticles prepared in example 1 of the present invention.
Fig. 3 is an energy spectrum analysis (EDS) diagram of gold and silver bimetallic nanoparticles prepared in example 1 of the present invention.
Fig. 4 is an X-ray diffraction (XRD) pattern of the gold and silver bimetallic nanoparticles prepared in example 1 of the present invention.
Fig. 5 is a Dynamic Light Scattering (DLS) diagram of gold and silver bimetallic nanoparticles prepared in example 1 of the present invention.
Fig. 6 is a graph showing the change of the decoloring rate of 100mg/mL active black treated by gold and silver bimetallic nanoparticles with different concentrations (2, 4,6, 10mg/mL) in example 2 of the invention.
Fig. 7 is a graph of the change in the decolorization rate of 100mg/mL safranin O treated with gold and silver bimetallic nanoparticles of different concentrations (2, 4,6, 10mg/mL) in inventive example 3.
Detailed Description
In order to better explain the present invention and to facilitate the understanding of the technical solutions of the present invention, the present invention is further described in detail below with reference to the specific embodiments and the accompanying drawings. It should be understood that the following examples are illustrative only and do not represent or limit the scope of the present invention, which is defined by the claims.
The reagents and instruments used in the following examples are not indicated by manufacturers, and are all conventional products available on the market.
The following examples use the customary writing in the field for the concentration units: m, mM or. mu.M, representing mol/L, mmol/L or. mu. mol/L, respectively.
Strains and culture conditions
Deinococcus radiodurans (Deinococcus radiodurans) purchased from American Type Culture Collection (ATCC) with accession number ATCC 13939.
TGY liquid medium: 0.5% peptone, 0.3% yeast extract and 0.1% Glucose (Glucose) were sterilized in an autoclave.
TGY solid medium: add 1.5% agar in TGY broth.
Example 1: preparation of gold and silver bimetallic nanoparticles
(1) Cell culture and collection: after activated culture of deinococcus radiodurans on TGY solid medium, single colonies were picked and cultured overnight in 5ml TGY liquid medium with shaking (32 ℃, 220 rpm). The culture solution was inoculated into 500ml of TGY broth at a ratio of 1:100 (by volume), and cultured at 32 ℃ and 220rpm to logarithmic phase (OD)6001.0). The cells were collected by centrifugation (8,000 Xg, 10min) and washed several times with 0.01M phosphate buffer, pH 7.0.
(2) Breaking the thallus and preparing a protein extract: the cells were resuspended in phosphate buffer and shaken well and disrupted with a cell disrupter (3min, 4 ℃, 1200 bar). The disruption solution was centrifuged (15,000 Xg, 30min), cell debris and non-disrupted cells were removed, and the supernatant was collected. To the supernatant was added ammonium sulfate solid so that the final concentration of ammonium sulfate was 80% (w/v) (i.e., the amount of ammonium sulfate was 80g per 100mL of the supernatant), and after standing at 4 ℃ for 4 hours for low-temperature salting out, the precipitate was collected by centrifugation at 12,000 Xg for 10 min. The collected precipitate was dissolved with ultrapure water, and the salt ion concentration in the solution was reduced by dialysis (dialysis for 24 hours or more with a cellulose acetate dialysis membrane having a cut-off of 14 kDa), and the dialyzed protein extract solution was concentrated to 2mg/mL (i.e., the protein concentration in the concentrated protein extract solution was 2mg/mL) by centrifugation at 4,800 Xg using an ultrafiltration concentration tube (available from Millipore Co., MA, USA)), and the concentrated protein extract solution was stored at 4 ℃ for subsequent use as a synthesizing agent.
(3) Synthesizing and purifying gold and silver bimetallic nanoparticles: adding Ag into the synthetic agent with the protein concentration of 2mg/ml obtained in the step (2)+Salt solution (silver nitrate, AgNO)3) Straight, straightTo Ag+At a final concentration of 1mM, reacting at 37 ℃ for 12 hours at pH 7.0, centrifuging at 20,000 Xg for 10min, and collecting the precipitate obtained by centrifugation; washing the precipitate with ultrapure water, centrifuging at 12,000 Xg for 10min, repeating the washing and centrifuging several times, re-suspending the precipitate with ultrapure water, and dialyzing with a cellulose acetate dialysis membrane with a cut-off of 14kDa for 24 hr or more to remove Ag+Then, the precipitate was centrifuged at 20,000 Xg for 10min, the precipitate obtained by the centrifugation was dissolved in ultrapure water, and Au was further added thereto3+Salt solution (chloroauric acid, HAuCl)4·3H2O) mixing thoroughly until Au3+Was 1mM, and reacted at 37 ℃ and pH 7.0. As the reaction proceeded, the solution changed from colorless upon initial mixing to purple.
(4) Purifying gold and silver bimetallic nanoparticles: after 6 hours of reaction, the reaction solution was filtered with a 0.22 μm syringe filter, the obtained filtrate was centrifuged at 20,000 Xg for 10min, the precipitate obtained by centrifugation was washed with ultrapure water and then centrifuged (20,000 Xg, 10min), after washing and centrifugation were repeated several times, the precipitate obtained by centrifugation was dissolved with ultrapure water and dialyzed with a cellulose acetate dialysis membrane having a cut-off of 14kDa for 24 hours or more to remove unreacted metal ions, and the solution after dialysis (referred to as dialysis solution for short) was freeze-dried to obtain the final product.
Characterization and identification:
for the dialyzed solution before the final product is obtained by freeze-drying in the above example 1, scanning is performed by using UV/Vis at the wavelength of 400-600nm to obtain a spectrum of an ultraviolet visible light absorption spectrum (UV/Vis), as shown in FIG. 1, a maximum absorption peak appears around 550nm, which means that gold and silver bimetallic nanoparticle characteristic substances in the final product are generated.
Scanning electron microscope-energy spectrum (SEM-EDS) analysis was performed on the final product prepared in example 1 above, and the scanning electron microscope results are shown in fig. 2. As can be seen from fig. 2, the final product nanoparticles prepared in example 1 were spherical and partially aggregated, because the surface of the particles was coated with proteins. The result of energy spectrum analysis (EDS) is shown in FIG. 3, the position of 2.1keV corresponds to the characteristic peak of Au nano-particles, and the position of 3keV corresponds to the characteristic peak of Ag nano-particles, thus proving the formation of gold and silver bimetallic nano-particles in the final product. As can be seen from FIG. 3, the spectra contain elements such as C, N, O, P, S in addition to the elements Au and Ag, indicating the presence of protein in the final product.
XRD analysis was performed on the final product prepared in example 1 above, and the result is shown in fig. 4. In fig. 4, diffraction peaks 32.32 °, 38.3 °, 46.38 °, 66.12 °, 77.7 °, and 85.66 ° correspond to crystal planes of [101], [111], [200], [220], [311], and [222] of face-centered cubic silver, respectively. And diffraction peaks 38.3 °, 44.56 °, 64.66 °, 77.7 °, and 81.9 ° correspond to the crystal planes of [111], [200], [220], [311], and [222] of face-centered cubic gold, respectively. The final product is shown to be a gold-silver bimetallic crystal structure.
The Dynamic Light Scattering (DLS) assay was performed on the dialyzed solution before the final product was obtained by freeze-drying in example 1, and the results are shown in FIG. 5, which shows that the particle size distribution of the final product in the solution was nearly monodisperse, the average size was 149.8nm in the average hydrated particle size, and the polydispersity index (PDI) was 0.397. + -. 0.021.
By combining the above characteristics and the identification results, it can be seen that: the final product obtained in the embodiment 1 is gold and silver bimetallic spherical nanoparticles with uniformly distributed particle sizes, and the surfaces of the particles are wrapped and modified by protein, namely, the gold and silver bimetallic spherical nanoparticles are protein-modified gold and silver bimetallic spherical nanoparticles, which are simply referred to as gold and silver bimetallic spherical nanoparticles.
Example 2: gold and silver bimetal nano-particles for degrading active black
The gold and silver bimetallic nanoparticles prepared in example 1 are used for degrading active black (active black-5 provided by Zhejiang lea soil chemical research institute is adopted), and the method comprises the following steps:
(1) dissolving 100mg of purified gold and silver bimetallic nanoparticles into 10ml of ultrapure water to obtain a gold and silver bimetallic nanoparticle solution with the final concentration of 0.01 g/ml.
(2) At normal temperature (25 ℃), the gold-silver bimetallic nanoparticle solution and the active black dye are fully mixed according to different proportions, and the volume is determined by ultrapure water, so that the final concentration of the active black dye is 100mg/L, and the final concentrations of the gold-silver bimetallic nanoparticle solution are respectively 2, 4,6, 8 and 10 mg/ml. The mixture was reacted at room temperature for 30min, centrifuged (20,000 Xg, 10min), and the supernatant was collected.
And (3) measuring the decoloring effect:
the characteristic absorption wavelength of the active black is 597nm, and the absorption value of the obtained supernatant is measured and analyzed by an enzyme-labeling instrument under the wavelength to determine the decolorization efficiency. The method comprises the following specific steps:
determining OD of supernatant obtained by treating active black with gold and silver double-metal nanoparticle solutions with different concentrations in the step (2) by using an enzyme-labeling instrument597The respective absorption values were obtained, and the decoloring ratios were calculated by the following formulas, and the corresponding relationship is shown in fig. 6. In fig. 6, the abscissa axis represents the concentration of the gold and silver bimetallic nanoparticle solution, and the ordinate axis represents the decolorization rate. The percentage of decolorization was calculated as follows:
decolorization ratio%f/Ai) X 100, wherein AfRefers to the final absorption value after treatment, AiRefers to the initial absorbance before treatment (control).
As can be seen from fig. 6: the decoloring effect of the gold-silver bimetallic nanoparticle solution on active black is increased along with the increase of the concentration of the gold-silver bimetallic nanoparticles, the active black dye with the final concentration of 100mg/L is treated by the gold-silver bimetallic nanoparticle solution with the final concentration of 10mg/ml, the decoloring rate is highest and reaches 94.7%, and the gold-silver bimetallic nanoparticles modified by protein functionalization can efficiently degrade the active black dye.
In addition, for the gold and silver bimetallic nanoparticle solutions with final concentrations of 2mg/mL and 4mg/mL, the absorption values at different times are respectively measured, and the decolorization rates at different times are calculated. And (3) displaying a calculation result: the two gold and silver bimetallic nanoparticle solutions with final concentrations can basically reach the corresponding highest decolorization rate within 5 min. The result shows that the gold-silver bimetallic nanoparticle solution can efficiently decolor the active black in a short time.
Aiming at the phenomenon that the active black reported in the literature has a composite color after decolorization, the invention also carries out corresponding experiments: after the decolored active black is placed in the air for a period of time, the phenomenon of color reversion of re-oxidation to blue does not occur. The reason for this is probably that the gold and silver bimetallic nano-particles of the invention have stronger degradation capability to the active black dye, and the dye is rapidly adsorbed on the gold and silver bimetallic nano-material with larger specific surface area and degraded in the treatment process.
Example 3: gold and silver bimetal nano-particles for degrading safranin O
The gold and silver bimetallic nanoparticles prepared in example 1 were used to degrade safranin O (available from Sigma-Aldrich Co. (MO, USA)) comprising the following steps:
(1) dissolving 100mg of purified gold and silver bimetallic nanoparticles into 10ml of ultrapure water to obtain a gold and silver bimetallic nanoparticle solution with the final concentration of 0.01 g/ml.
(2) At normal temperature (25 ℃), the gold-silver bimetallic nanoparticle solution and the safranin O dye are fully mixed according to different proportions, and the volume is determined by ultrapure water, so that the final concentration of the safranin O dye is 100mg/L, and the final concentrations of the gold-silver bimetallic nanoparticle solution are respectively 2, 4,6, 8 and 10 mg/ml. The mixture was reacted at room temperature for 30min, centrifuged (20,000 Xg, 10min), and the supernatant was collected.
And (3) measuring the decoloring effect:
determining OD of supernate obtained by treating safranine O with gold and silver bimetallic nanoparticle solutions with different concentrations in the step (2) by using an enzyme-labeling instrument510And obtaining the corresponding absorption value. The decolorization ratio was calculated by the following formula, and the correspondence between the concentration and the decolorization ratio is shown in FIG. 7. In fig. 7, the abscissa axis represents the concentration of the gold and silver bimetallic nanoparticle solution, and the ordinate axis represents the decolorization rate. The percentage of decolorization was calculated as follows:
decolorization ratio%f/Ai) X 100, wherein AfRefers to the final absorption value after treatment, AiRefers to the initial absorbance before treatment (control).
As can be seen from fig. 7: the decoloring rate of the gold and silver bimetallic nanoparticle solution on safranin O can reach more than 50%, which shows that the gold and silver bimetallic nanoparticles modified by protein functionalization can effectively degrade safranin O dye.
It will thus be seen that the objects of the invention have been fully and effectively accomplished. The method and principles of the present invention have been shown and described in the examples, which can be modified in any way without departing from the principles. Therefore, this invention includes all modifications encompassed within the spirit and scope of the claims.
Claims (10)
1. A synthetic method of functionalized modified metal nanoparticles comprises the following steps:
(1) crushing deinococcus radiodurans under pressure, collecting supernatant, and preparing protein extract by salting out method as synthesis agent;
(2) adding Ag to the synthesis agent+After the salt solution reacts, Au is added3+And (3) salt solution reaction to synthesize the protein-modified gold-silver bimetallic nano-particles.
2. The method of synthesizing the functionally modified metal nanoparticle of claim 1, comprising the steps of:
(1) resuspending deinococcus radiodurans in logarithmic phase with phosphate buffer solution, sufficiently oscillating, performing cell disruption at 4 deg.C under 1200bar for 3min, centrifuging the disruption solution at 15,000 Xg for 30min, and collecting supernatant; adding ammonium sulfate into the supernatant according to the proportion of 80g of ammonium sulfate to every 100mL of supernatant, standing for 4 hours at 4 ℃, centrifuging for 10min at 12,000 Xg, and collecting the precipitate obtained by centrifugation; dissolving the precipitate with ultrapure water, dialyzing with a cellulose acetate dialysis membrane with a cut-off of 14kDa for over 24 hours, and concentrating the dialyzed solution to a protein concentration of 2mg/ml to obtain a protein extract solution as a synthesis agent;
(2) adding Ag to the synthesis agent+Salt solution to Ag+Reacting at a final concentration of 1mM at 37 deg.C and pH 7.0 for 12 hr or more, centrifuging at 20,000 Xg for 10min, and collecting the precipitate; washing the precipitate with ultrapure water, centrifuging for 10min at 12,000 Xg, repeating the washing and centrifuging for several times, resuspending the precipitate with ultrapure water, dialyzing with 14kDa cellulose acetate dialysis membrane for over 24 hr, and separating at 20,000 XgCentrifuging for 10min, dissolving the precipitate in ultrapure water, and adding Au3+The salt solution was mixed well until Au3+The final concentration of the gold and silver nanoparticles is 1mM, and the reaction is carried out for more than 2 hours under the conditions of pH 7.0 and 37 ℃ to obtain a reaction solution containing the protein-modified gold and silver bimetallic nanoparticles.
3. The method of synthesizing the functionally modified metal nanoparticle of claim 2, further comprising: and purifying the reaction solution to obtain the gold and silver bimetallic nanoparticles modified by the protein.
4. The method of claim 3, wherein the purification step comprises the steps of: filtering the reaction solution by using a 0.22 mu m needle filter, centrifuging 20,000 Xg of obtained filtrate for 10min, washing the obtained precipitate by centrifugation with ultrapure water, centrifuging for 10min again by using 20,000 Xg, repeatedly washing and centrifuging for a plurality of times, dissolving the precipitate by using the ultrapure water, centrifuging for more than 24 hours by using a cellulose acetate dialysis membrane with the interception amount of 14kDa, and freeze-drying the solution after dialysis to obtain the protein-modified gold and silver bimetallic nanoparticles.
5. The method for synthesizing functionally modified metal nanoparticles of claim 2, wherein in step (1), the concentration is performed by centrifugation at 4,800 Xg using an ultrafiltration concentration tube.
6. The method for synthesizing functionally modified metal nanoparticles of claim 1 or 2, wherein in the step (2), Ag is added+The salt solution is silver nitrate solution, Au3+The salt solution is chloroauric acid solution.
7. The method of claim 2, wherein step (2) is performed with Ag+The salt solution was reacted for 12 hours with Au3+The salt solution reaction time was 6 hours.
8. The protein-modified gold-silver bimetallic nanoparticle prepared by the synthesis method of any one of claims 1-7.
9. The protein-modified gold-silver bimetallic nanoparticle of claim 8 for degrading reactive black dye.
10. The protein-modified gold-silver bimetallic nanoparticle of claim 8 for degrading safranin O dye.
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