CN105112453A - Preparation method of nano-particle material - Google Patents
Preparation method of nano-particle material Download PDFInfo
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- CN105112453A CN105112453A CN201510601633.2A CN201510601633A CN105112453A CN 105112453 A CN105112453 A CN 105112453A CN 201510601633 A CN201510601633 A CN 201510601633A CN 105112453 A CN105112453 A CN 105112453A
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Abstract
The invention provides a culture medium prepared from peptone, yeast and NaCl, wherein strains cultured in the culture medium are pantoea agglomerans belonging to IMH cells. After being collected, thallus cells are dispersed to different cultivation systems according to the difference among nano-particles required to be synthesized, metal ions are added to meet the requirement of a proper concentration, reactions are carried out for several hours, and separation and collection are carried out according to different reaction times, so as to obtain metal nano-particle materials of different particle diameters. The whole process is environmentally friendly, simple and convenient to operate, and low in cost; the obtained nano-particle materials have wide application prospects in the aspects of catalysis, adsorption, detection and the like.
Description
Technical field
The invention provides a kind of preparation method of nano-particle material, belong to nano material synthesis technical field.
Technical background
Nano material refers to the material with property having at least one dimension to be in nanoscale scope or to be made up of as elementary cell them in three dimensions.Due to when material particle size enters nanometer scale, itself there is quantum size effect, small-size effect, surface effects, quantum tunneling effect, thus show physics and the chemical property of many uniquenesses, have broad application prospects at numerous areas such as catalysis, environmental protection, the energy, medicine.
At present, the synthetic method of nano particle can be divided into generally: " from top to bottom " and " from bottom to top " two kinds of approach." from top to bottom " method is a process from large to small, and lump is broken by physical method, and pulverize, the modes such as grinding, as high-energy ball milling method, change the particle of nanometer scale into.These class methods are simple to operate, and cost is lower, suitability for scale production.But obtained material granule is comparatively large, and size distribution is uneven, and shape is difficult to control." from bottom to top " method is by suitable chemical reaction, and the atom in chemical reaction between material must carry out group row, and from molecule, atom sets out and prepares nano particle.As methods such as the precipitator method, sol-gel method, hydro-thermal solvent-thermal method, microemulsion method, template.Chemical synthesis process is versatile and flexible, can be used for the nano particle preparing different morphologies, but most chemical method needs to introduce more chemical reagent, may bring certain problem of environmental pollution.
Therefore, the chemical research prepared along with material is increasingly active, and biosynthetic method causes the attention of researchist gradually.The conversion process of biological mediation does not limit by harsh conditions such as pressure, temperature, pH, can carry out under mild conditions.Therefore organism such as bacterium, fungi, algae etc. or biomass reduction is utilized to prepare nano particle, not only can obtain some results that Physical and chemical method are difficult to obtain, and process does not need to introduce other chemical reagent, biological character itself can also be made full use of.So the method has more the feature of Sustainable development, become a focus in nano particle preparation research field.Except having bioactive molecule as except reductive agent synthetic metals nanoparticle approach, PNAS reported first in 1999 Pseudomonas stutzeri preparing silver nano granules by reducing, opens novel method prepared by metal nanoparticle-utilize microorganism in situ to reduce.
Utilize microorganism in situ synthetic metals nano particle to refer to that metal ion utilizes the effect of reductase enzyme in cell surface functional group or born of the same parents, be reduced to the metal nanoparticle of zeroth order.At present, the research of machine-processed aspect is divided into two aspects according to method difference, and one is the carboxyl of cytolemma or cell wall, and amide group, ketone group and aldehyde radical can be used as the electron donor of metal ion.The active organo-functional group of these cell surfaces can adsorb, complexing and chelated metal ions, and in-situ reducing becomes metallic particles on cell walls, somatic cells wall surface and metallic particles produce and interact simultaneously, stop it to move and reduce reunion, thus obtain metal nanoparticle.Two is enzyme catalysis reduction processes, the enzyme that microorganism produces generates in the process of nano particle in reduction and plays the effect of catalyzer, utilizes electron transit mediator to be reduced to metal ion by the transfer transport of reducing substances, thus synthetic metals nano particle.
Utilize microorganism in situ to reduce and prepare the deficiency that metal nanoparticle not only can improve physico-chemical processes in preparation method, nano particle diameter is simultaneously homogeneous, stability is high, and has good biocompatibility, thus has greatly expanded the application of nano particle.The present invention have developed a kind of novel method utilizing microorganism in situ reduction synthesis metal nanoparticle, the present invention utilizes the bacterial strain of extraction purification in the actual soil in Shanxi Province, through series of experiments, gold and silver, palladium, selenium four kinds of metal nanoparticles are successfully prepared, method of the present invention is to biological detection, and the fields such as bio-imaging have great importance.
Summary of the invention
The object of the invention is to utilize special microbial strains in-situ reducing synthetic metals nano particle, the method can apply to the synthesis of multiple elemental metals nano particle.
The present invention utilizes peptone, yeast and sodium-chlor to prepare substratum, cultivates bacterial strain pantoea agglomerans and belongs to IMH cell.Collect after being cultured to exponential phase cells and obtain cell.Afterwards by culture systems extremely different for cell dispersal, add the metal ion of proper concn.After stoichiometric number hour, separable collection obtains the metal nanoparticle of different-grain diameter.
According to a specific embodiment of the present invention, a kind of preparation method of nano-particle material, it comprises the steps:
(1) substratum preparation: take 1g peptone, 0.5g yeast and 1g sodium-chlor and be dissolved in 100mL deionized water, stir 3 ~ 10 minutes, blended solid is melted completely, forms uniform solution, and adjusted to ph to 7;
(2) medium sterilization: the substratum prepared is placed in Autoclave, at 120 DEG C, high-temperature sterilization 20min, afterwards cooling preservation under room temperature;
(3) bacterial strain activation and inoculation: with transfering loop, bacterial strain pantoea agglomerans is belonged to IMH and access sterilized culture medium solution, solution is placed in shaking table, at 30 DEG C, under 150rpm condition, 12h is cultivated in concussion, after drawing 1mL activation subsequently, bacterium liquid is forwarded to the fresh culture medium solution obtained, again solution is placed in shaking table, at 30 DEG C, under 150rpm condition, 12h is cultivated in concussion; .
(4) cell harvesting: by the bacterium liquid that obtains in step (3) under sterilized atmosphere, be transferred in centrifuge tube is under the condition of 8000rpm centrifugal 5 minutes at rotating speed.After centrifugal end, abandoning supernatant, collects bottom thalline in centrifuge tube.
(5) silver-colored in-situ reducing: by the thalline (OD obtained in step (4)
600=1.0) be dispersed to and remove in the LB substratum of NaCl, add the silver ions that concentration is 1mM, reaction 8h, obtains the silver nano-grain that particle diameter is 100nm.
(6) golden in-situ reducing: by the thalline (OD obtained in step (4)
600=1.0) be dispersed in PBS damping fluid, add the gold ion that concentration is 1.8mM, reaction 4h, obtains the gold nano grain that particle diameter is 30nm.
(7) palladium in-situ reducing: by the thalline (OD obtained in step (4)
600=1.0) be dispersed in PBS damping fluid, add the palladium ion that concentration is 2mM, reaction 8h, obtains the palladium nano-particles that particle diameter is 50nm.
(8) selenium in-situ reducing: by the thalline (OD obtained in step (4)
600=1.0) be dispersed in LB substratum, add the plasma selenium that concentration is 2mM, reaction 4h, obtains the palladium nano-particles that particle diameter is 80nm.
Accompanying drawing explanation
Below by marginal data principal character of the present invention.
The transmission electron microscope photo of the silver nano-grain that accompanying drawing 1 obtains for the present invention, experimental result shows that the silver nano-grain size synthesized is even, and particle diameter is about 100nm.
The transmission electron microscope photo of the gold nano grain that accompanying drawing 2 obtains for the present invention, experimental result shows that the gold nano grain size synthesized is even, and particle diameter is about 30nm.
The transmission electron microscope photo of the palladium nano-particles that accompanying drawing 3 obtains for the present invention, experimental result shows that the palladium nano-particles size synthesized is even, and particle diameter is about 50nm.
The transmission electron microscope photo of the selenium nano particle that accompanying drawing 4 obtains for the present invention, experimental result shows that the selenium nano particle size synthesized is even, and particle diameter is about 80nm.
Inventive embodiments
The present invention is set forth below further by embodiment.
Embodiment 1 silver nano-grain
Taking 10g peptone, 5g yeast and 10g sodium-chlor is dissolved in 1L deionized water, and adjust ph is about 7.Preserve after autoclave sterilization.With transfering loop, bacterial strain pantoea agglomerans is belonged to IMH to access in the 100mL culture medium solution obtained.Afterwards solution is placed in shaking table, at 30 DEG C, under 150rpm condition, 12h is cultivated in concussion.Subsequently, after drawing 1mL activation, bacterium liquid is forwarded in the substratum of sterilized 100mL, again solution is placed in shaking table, and at 30 DEG C, under 150rpm condition, 12h is cultivated in concussion.Get 50mL logarithmic phase bacterium liquid in centrifuge tube, under 8000rpm condition centrifugal 5 minutes.After end, remove supernatant liquor, the cell of collection tube inner bottom part.Remove collecting the cell dispersal obtained in the LB substratum of NaCl to 50mL is sterilized.Add a certain amount of silver nitrate solution, make the ultimate density of silver ions in solution reach 1mM.Mixing solutions is placed in shaking table, at 30 DEG C, after cultivating 8h under the condition of 150rpm, can silver nano-grain be obtained.
Embodiment 2 gold nano grain
Taking 10g peptone, 5g yeast and 10g sodium-chlor is dissolved in 1L deionized water, and adjust ph is about 7.Preserve after autoclave sterilization.With transfering loop, bacterial strain pantoea agglomerans is belonged to IMH to access in the 100mL culture medium solution obtained.Afterwards solution is placed in shaking table, at 30 DEG C, under 150rpm condition, 12h is cultivated in concussion.Subsequently, after drawing 1mL activation, bacterium liquid is forwarded in the substratum of sterilized 100mL, again solution is placed in shaking table, and at 30 DEG C, under 150rpm condition, 12h is cultivated in concussion.Get 50mL logarithmic phase bacterium liquid in centrifuge tube, under 8000rpm condition centrifugal 5 minutes.After end, remove supernatant liquor, the cell of collection tube inner bottom part.Cell dispersal collection obtained is in the sterilized PBS damping fluid of 50mL.Add a certain amount of chlorauric acid solution, make the ultimate density of GOLD FROM PLATING SOLUTION ion reach 1.8mM.Mixing solutions is placed in shaking table, at 30 DEG C, after cultivating 4h under the condition of 150rpm, can gold nano grain be obtained.
Embodiment 3 palladium nano-particles
Taking 10g peptone, 5g yeast and 10g sodium-chlor is dissolved in 1L deionized water, and adjust ph is about 7.Preserve after autoclave sterilization.With transfering loop, bacterial strain pantoea agglomerans is belonged to IMH to access in the 100mL culture medium solution obtained.Afterwards solution is placed in shaking table, at 30 DEG C, under 150rpm condition, 12h is cultivated in concussion.Subsequently, after drawing 1mL activation, bacterium liquid is forwarded in the substratum of sterilized 100mL, again solution is placed in shaking table, and at 30 DEG C, under 150rpm condition, 12h is cultivated in concussion.Get 50mL logarithmic phase bacterium liquid in centrifuge tube, under 8000rpm condition centrifugal 5 minutes.After end, remove supernatant liquor, the cell of collection tube inner bottom part.Cell dispersal collection obtained is in the sterilized PBS damping fluid of 50mL.Add a certain amount of palladium nitrate solution, make the ultimate density of palladium ion in solution reach 2mM.Mixing solutions is placed in shaking table, at 30 DEG C, after cultivating 8h under the condition of 150rpm, can palladium nano-particles be obtained.
Embodiment 4 selenium nano particle
Taking 10g peptone, 5g yeast and 10g sodium-chlor is dissolved in 1L deionized water, and adjust ph is about 7.Preserve after autoclave sterilization.With transfering loop, bacterial strain pantoea agglomerans is belonged to IMH to access in the 100mL culture medium solution obtained.Afterwards solution is placed in shaking table, at 30 DEG C, under 150rpm condition, 12h is cultivated in concussion.Subsequently, after drawing 1mL activation, bacterium liquid is forwarded in the substratum of sterilized 100mL, again solution is placed in shaking table, and at 30 DEG C, under 150rpm condition, 12h is cultivated in concussion.Get 50mL logarithmic phase bacterium liquid in centrifuge tube, under 8000rpm condition centrifugal 5 minutes.After end, remove supernatant liquor, the cell of collection tube inner bottom part.Cell dispersal collection obtained is in the sterilized LB substratum of 50mL.Add a certain amount of sodium selenite solution, make the ultimate density of plasma selenium in solution reach 2mM.Mixing solutions is placed in shaking table, at 30 DEG C, after cultivating 4h under the condition of 150rpm, selenium nano particle can be obtained.
Claims (6)
1. a nanometer grain preparation method, is characterized in that carrying out according to following step:
(1) bacterial strain activation and inoculation: with transfering loop, bacterial strain pantoea agglomerans is belonged to IMH and access sterilized culture medium solution, solution is placed in shaking table, at 30 DEG C, under 150rpm condition, 12h is cultivated in concussion, after drawing 1mL activation subsequently, bacterium liquid is forwarded to the fresh culture medium solution obtained, again solution is placed in shaking table, at 30 DEG C, under 150rpm condition, 12h is cultivated in concussion;
(2) cell harvesting: by the bacterium liquid that obtains in step (1) under sterilized atmosphere, be transferred in centrifuge tube, be under the condition of 8000rpm centrifugal 5 minutes at rotating speed, abandoning supernatant after centrifugal end, collects bottom thalline in centrifuge tube;
(3) silver-colored in-situ reducing: the thalline obtained in step (2) is dispersed to and removes in the LB substratum of NaCl, add the silver ions that concentration is 1mM, reaction 8h, obtains the silver nano-grain that particle diameter is 100nm;
(4) golden in-situ reducing: be dispersed in PBS damping fluid by the thalline obtained in step (2), adds the gold ion that concentration is 1.8mM, and reaction 4h, obtains the gold nano grain that particle diameter is 30nm;
(5) palladium in-situ reducing: be dispersed in PBS damping fluid by the thalline obtained in step (2), adds the palladium ion that concentration is 2mM, and reaction 8h, obtains the palladium nano-particles that particle diameter is 50nm;
(6) selenium in-situ reducing: be dispersed in LB substratum by the thalline obtained in step (2), adds the plasma selenium that concentration is 2mM, and reaction 4h, obtains the palladium nano-particles that particle diameter is 80nm.
2. a kind of nanometer grain preparation method as claimed in claim 1, in step (1), the compound method of substratum is: take 1g peptone, 0.5g yeast and 1g sodium-chlor are dissolved in 100mL deionized water, and adjust ph is 7.
3. a kind of nanometer grain preparation method as claimed in claim 1, in step (1), sterilising method is: the substratum prepared is placed in Autoclave, at 120 DEG C, high-temperature sterilization 20min, afterwards cooling preservation under room temperature.
4. a kind of nanometer grain preparation method as claimed in claim 1, the thalline OD obtained
600=1.0.
5. a kind of nanometer grain preparation method as claimed in claim 1, in step (3), the compound method removing the LB substratum of NaCl is: 1g peptone and 0.5g yeast are dissolved in 100mL deionized water, and adjust ph is 7.
6. a kind of nanometer grain preparation method as claimed in claim 1, in step (4-5), the compound method of PBSbuffer is: take potassium primary phosphate 0.27g, Sodium phosphate dibasic 1.42g, sodium-chlor 8g and Repone K 0.2g, add deionized water and be about the abundant stirring and dissolving of 800mL, then add concentrated hydrochloric acid and adjust pH to 7.4, be settled to 1L.
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Cited By (7)
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| CN105780067A (en) * | 2016-02-01 | 2016-07-20 | 中国科学院生态环境研究中心 | Method for in-situ synthesis of three-dimensional nanometer palladium catalyst layer through electrode activity biological membrane and application |
| CN107641632A (en) * | 2017-10-18 | 2018-01-30 | 福州大学 | A kind of method with the carbon-based point of Microbe synthesis |
| EP3330227A1 (en) * | 2016-11-30 | 2018-06-06 | Advanced-Connectek Inc. | Bacterially induced crystal particle, thermal conductive material and method for manufacturing bacterially induced crystal particle |
| CN109136128A (en) * | 2018-08-23 | 2019-01-04 | 国家海洋局第海洋研究所 | It is a kind of pair coccus and its screening technique and synthesis silver nano-grain in application |
| CN109295107A (en) * | 2018-07-14 | 2019-02-01 | 吉林市赛恩思纳米材料有限公司 | A kind of green synthesis method of metal nanoparticle |
| CN112573491A (en) * | 2020-12-18 | 2021-03-30 | 电子科技大学 | Coral-like Pd4Method for producing Se compound |
| CN114100676A (en) * | 2020-08-26 | 2022-03-01 | 浙江大学 | Synthesis method and application of functionalized modified metal nanoparticles |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105780067A (en) * | 2016-02-01 | 2016-07-20 | 中国科学院生态环境研究中心 | Method for in-situ synthesis of three-dimensional nanometer palladium catalyst layer through electrode activity biological membrane and application |
| EP3330227A1 (en) * | 2016-11-30 | 2018-06-06 | Advanced-Connectek Inc. | Bacterially induced crystal particle, thermal conductive material and method for manufacturing bacterially induced crystal particle |
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| US11359215B2 (en) | 2016-11-30 | 2022-06-14 | Acon-Holding Inc. | Bacterially induced crystal particle, thermal conductive material and method for manufacturing bacterially induced crystal particle |
| CN107641632A (en) * | 2017-10-18 | 2018-01-30 | 福州大学 | A kind of method with the carbon-based point of Microbe synthesis |
| CN109295107A (en) * | 2018-07-14 | 2019-02-01 | 吉林市赛恩思纳米材料有限公司 | A kind of green synthesis method of metal nanoparticle |
| CN109136128A (en) * | 2018-08-23 | 2019-01-04 | 国家海洋局第海洋研究所 | It is a kind of pair coccus and its screening technique and synthesis silver nano-grain in application |
| CN114100676A (en) * | 2020-08-26 | 2022-03-01 | 浙江大学 | Synthesis method and application of functionalized modified metal nanoparticles |
| CN112573491A (en) * | 2020-12-18 | 2021-03-30 | 电子科技大学 | Coral-like Pd4Method for producing Se compound |
| CN112573491B (en) * | 2020-12-18 | 2022-05-03 | 电子科技大学 | Coral-like Pd4Method for producing Se compound |
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