CN103170641B - Method of preparing platinum nano particles by using cotton bollworm karyotype polyhedrin - Google Patents
Method of preparing platinum nano particles by using cotton bollworm karyotype polyhedrin Download PDFInfo
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- CN103170641B CN103170641B CN201310087213.8A CN201310087213A CN103170641B CN 103170641 B CN103170641 B CN 103170641B CN 201310087213 A CN201310087213 A CN 201310087213A CN 103170641 B CN103170641 B CN 103170641B
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Abstract
Provided is a method of preparing platinum nano particles by using cotton bollworm karyotype polyhedrin. Inherent structural features of the cotton bollworm karyotype polyhedrin are used, a PtC14 solution is added to a purified cotton bollworm karyotype polyhedral body solution, and the precious metal platinum nano particles which are even in distribution and consistent in diameter of the particles are obtained on the surface of the cotton bollworm karyotype polyhedrin through hatch, ultrasonic wave assistance reduction treatment. Due to the fact that a cotton bollworm karyotype polyhedron carrier has natural specific structures of functional group repetitive distribution, the complex preparation technology of a conventional carrier is avoided, and therefore production cost is reduced. The sizes and distribution of the precious metal platinum nano particles are effectively controlled through control of the alkaline hydrolysis degree of the cotton bollworm karyotype polyhedron, and the method of preparing the platinum nano particles by using the cotton bollworm karyotype polyhedrin can be significantly applied to catalytic fuel cells or biosensing or the like.
Description
Technical field
The present invention relates to a kind of preparation method of nano material, particularly prepare the method for noble metal platinum nano particle.
Background technology
Noble metal nanometer material is owing to having the features such as good stability, small-size effect, skin effect, optical effect and good biocompatibility, in fields such as optics, sensor, catalysis and biological detection analyses, there is important using value, thus more and more get more and more people's extensive concerning.Size, pattern and composite construction thereof that Recent study focus concentrates on by controlling nano particle prepare regulatable novel nano particle.The preparation method of current noble metal platinum nano particle is a lot, but will obtain structure, form, size adjustable, is evenly distributed, stability good noble metal platinum nano particle is still more difficult.
In recent years, people often adopt the material with carbon element of different shape or other templates as carrier to control growth and the distribution of small particle diameter noble metal platinum nano particle.Such as, people (the ACS.Appl.Mate.Interfaces.2011 such as Ping Lin K μ o, 3,115-118) and the people (Chem.Mater.2006 such as Robert V H μ ll, 18,1780-1788) report the nano platinum particle utilizing porous carbon nanotube and multi-walled carbon nano-tubes preparation dispersion respectively, because the material with carbon element itself as load lacks activated adoption group, in preparation process, need the process through early stage to introduce the group that can be combined with noble metal.These operations make complicated process of preparation, improve production cost, and the distribution of the metallic obtained have very large randomness.Dan Wen etc. is published in the article of " physical chemistry C " (J.Phys.Chem.C.2009,113,13023-13028) and reports TiO
2the preparation of colloidal spheres supporting Pt nano particle, the product of preparation may be used for biology sensor material.But, due to TiO
2itself lacks adsorption group, is difficult to make noble metal nano particles be attached to its surface, needs to add aminopropyl trimethoxysilane to introduce NH
2-tiO is connected as intermediate
2with noble metal.
Summary of the invention
The object of the present invention is to provide a kind of distribution that effectively can be controlled noble metal platinum nano particle by the alkaline hydrolysis degree controlling albumen, preparation condition is gentle, productive rate is high, technique is simple, and the bioprotein nanostructured that the method directly adopts nature to exist is as the carrier of noble metal platinum nano particle, wide material sources, repeatability is high, a kind of method utilizing bollworm polyhedrosis albumen to prepare nano platinum particle that production cost is low.
The present invention mainly adopts natural bollworm polyhedrosis albumen to control the noble metal platinum nano particle of synthesis high degree of dispersion as carrier.First the present invention carries out purifying alkaline hydrolysis to bollworm polyhedrosis, obtains polyhedrin sheet, then adds PtCl
4solution carries out common hatching, on the avtive spot of the particular functional group's (as mercapto functional group) making precious metal ion be adsorbed onto polyhedrosis protein surface to contain, by the process of ultrasonic wave added coreduction after hatching special time, arrangement is just defined in order, the precious metals pt nano particle of size uniform at protein surface.
Technical scheme of the present invention specifically comprises the steps:
(1) purifying of bollworm polyhedrosis and alkaline hydrolysis:
Former for 0.5 ~ 1.0g heliothis armigera nuclear polyhedrosis virus powder is dissolved in 60 ~ 100mL ultra-pure water, its dissolving of ultrasonic promotion.The present invention's ultra-pure water used is prepared by ultra-pure water instrument.Get supernatant after the centrifugal 5 ~ 10min of 500 ~ 1200r/min, get precipitation after the centrifugal 20 ~ 30min of 3000 ~ 5000r/min, precipitation ultra-pure water is resuspended.Low speed like this, centrifugal 3 ~ 5 times repeatedly at a high speed, to last ultracentrifugal supernatant achromaticity and clarification, namely obtains purer polyhedral body.Gained polyhedral body is resuspended with the ultra-pure water that 1.5 ~ 2.5mL is cold, add 150 ~ 250 μ L alkali solution liquid process 10 ~ 25min.Above-mentioned alkali solution liquid is 0.3mol/L sodium carbonate, 0.5mol/L sodium chloride and 0.03mol/L disodium ethylene diamine tetra-acetic acid solution.Then regulate pH to 7 ~ 8 with the glacial acetic acid of 0.5mol/L, centrifugal 2 ~ 3 times of 5000 ~ 7000r/min, gets supernatant.After at 4 ~ 10 DEG C, ultracentrifugation 1 ~ 2h under 25000 ~ 30000r/min, precipitates spot 1 ~ 2mL ultra-pure water resuspended by gained after abandoning supernatant.
(2) the noble metal platinum nano particle of bollworm polyhedrosis protein load is prepared:
It is the PtCl of 1 ~ 10mM that the bollworm polyhedrosis protein liquid getting 100 ~ 300 μ L alkaline hydrolysis process adds 100 ~ 300 μ L concentration
4in solution, fully mix, in 20 ~ 30 DEG C, shaking table hatching 15 ~ 25h under 130 ~ 170r/min.Then dropwise adding the concentration that 50 ~ 100 μ L newly prepare is the NaBH of 5 ~ 10mM
4solution, limit ultrasonic limit reduction.In 20 ~ 30 DEG C after reduction, under 130 ~ 170r/min, shaking table hatching 3 ~ 6h, namely obtains bollworm polyhedrosis protein load noble metal platinum nano particle.
Because bioprotein comprises various active group, amino acid sequence arranging rule, biodegradable, bio-compatible, has and improve and strict molecular recognition function; in the preparation of noble metal platinum nano particle, both can be used as the noble metal platinum nano particle that protective agent or adjusting control agent prepare different size, pattern, can be used as again the regularly arranged noble metal platinum nano particle of guiding agent.Simultaneously the architectural feature of biomolecule self and space confinement effect thereof can carry out accuracy controlling to the synthesis of nano particle, thus obtain the nano particle with desired size size and arranging rule.The bollworm polyhedrosis albumen that the present invention adopts is that heliothis armigera nuclear polyhedrosis virus produces in later period of infection great expression, and self forms the supramolecular protein crystal of compact structure.Bollworm polyhedrosis albumen is made up of a kind of trimerical subunit structure of densification, connected between tripolymer by disulfide bond.The theoretical length of a repetitive of this protein structures is 10nm, and each tripolymer length is 5nm.The arrangement of this specific structure repeatability, its outer surface is rich in a large amount of amino acid, and contained active function groups can adsorb many kinds of metal ions to its surface.And this albumen distinguishing feature is exactly by regulating the alkaline hydrolysis degree of polyhedrosis alkaline hydrolysis time Function protein, and then can also control the distribution of noble metal platinum nano particle.
Described noble metal platinum nano particle has following architectural feature: bollworm polyhedrosis protein structure rule, individual layer albumen presents orderly fenestral fabric, and protein surface has specific avtive spot to be easily combined with metal ion.Gained noble metal platinum nano particle is evenly distributed on protein surface, and decentralization is high, and size, the decentralization of particle are consistent with bollworm polyhedrosis albumen.
This method not only decreases the preparation technology of traditional nano platinum particle carrier complexity, and without the need to introducing external source active group, namely the active function groups that albumen itself contains controls the size of nano platinum particle as adjusting control agent, control again arrangement and the decentralization of nano platinum particle as guiding agent.
The invention has the beneficial effects as follows: directly adopt natural bollworm polyhedrosis albumen as carrier, any specially treated need not be carried out, through hatching altogether with material system, the process of ultrasonic wave added coreduction, can obtain the noble metal platinum nano particle of bollworm polyhedrosis protein load.The noble metal platinum nano particle regulating and controlling to grow as carrier with existing material with carbon element and porous oxide is compared, the noble metal platinum nano particle that the present invention obtains has that size is little, domain size distribution evenly, dispersion advantages of higher.This preparation method's technique is simple, mild condition, environment-friendly high-efficiency, and carrier wide material sources are easy to get, with low cost, is easy to realize large-scale production.
Accompanying drawing explanation
Below in conjunction with drawings and Examples, the invention will be further elaborated.
Fig. 1 is the TEM figure of the bollworm polyhedrosis albumen that alkaline hydrolysis 15min obtains;
Fig. 2 is the TEM figure of the nano platinum particle of the bollworm polyhedrosis protein load of the alkaline hydrolysis 15min of preparation;
Fig. 3 is the TEM figure of the bollworm polyhedrosis albumen that alkaline hydrolysis 25min obtains;
Fig. 4 is the TEM figure of the nano platinum particle of the bollworm polyhedrosis protein load of the alkaline hydrolysis 25min of preparation.
Detailed description of the invention
Embodiment 1
Former for 0.5g heliothis armigera nuclear polyhedrosis virus powder is dissolved in 60mL ultra-pure water, its dissolving of ultrasonic promotion.Prepared by the PL5124-PALL model ultra-pure water instrument that ultra-pure water used is produced by pure water system Co., Ltd of Vivendi of High Wycombe city of Britain.Get supernatant after the centrifugal 10min of 1000r/min, get precipitation after the centrifugal 30min of 4000r/min, precipitation ultra-pure water is resuspended.Low speed like this, centrifugal 4 times repeatedly at a high speed, to last ultracentrifugal supernatant achromaticity and clarification, namely obtains purer polyhedral body.By resuspended for ultra-pure water cold for gained polyhedral body 2mL, add 200 μ l alkali solution liquid process 15min.Then get supernatant after regulating the centrifugal 5min of pH to 7,5000r/min with the glacial acetic acid of 0.5mol/L, 6000r/min is centrifugal, and 5min gets supernatant, and the centrifugal 5min of 7000r/min, gets supernatant.After at 4 DEG C, ultracentrifugation 1h under 28000r/min, precipitates spot 1.5mL ultra-pure water resuspended by gained after abandoning supernatant.Observe its pattern under transmission electron microscope and see Fig. 1.Fig. 1 is the TEM figure of the bollworm polyhedrosis albumen after embodiment 1 purifying and alkaline hydrolysis, as can be seen from the figure the bollworm polyhedrosis albumen of gained presents the fenestral fabric of individual layer, single lattice are that a tripolymer assumes diamond in shape, be approximately square, size is 5nm, multiple tripolymer repeated arrangement forms reticuloprotein sheet, and the size of albumen flakes is 20 ~ 80nm, and mainly concentrating is 40 ~ 50nm.Each tripolymer has active function groups, and for the forming core of noble metal provides site, the spacing effect in its space limits the particle diameter of noble metal platinum nano particle is simultaneously 4 ~ 5nm.
It is the PtCl of 5mM that the bollworm polyhedrosis protein liquid getting 200 μ L alkaline hydrolysis process adds 100 μ L concentration
4solution, fully mixes, in 25 DEG C, and shaking table hatching 24h under 140r/min.Then dropwise adding the concentration that 75 μ L newly prepare is the NaBH of 5mM
4solution, limit ultrasonic limit reduction.In 25 DEG C after reduction, under 140r/min, shaking table hatching 6h, namely obtains the nano platinum particle of bollworm polyhedrosis protein load.Observe its pattern under transmission electron microscope and see Fig. 2.Fig. 2 is the TEM figure of the nano platinum particle of bollworm polyhedrosis load prepared by embodiment 1, can disperse uniformly the nano platinum particle of gained from figure, size and shape and polyhedrosis albumen basically identical, single nano particle diameter is 4 ~ 5nm, does not have agglomeration.
Embodiment 2
Former for 0.5g heliothis armigera nuclear polyhedrosis virus powder is dissolved in 60mL ultra-pure water, its dissolving of ultrasonic promotion.Prepared by the PL5124-PALL model ultra-pure water instrument that ultra-pure water used is produced by pure water system Co., Ltd of Vivendi of High Wycombe city of Britain.Get supernatant after the centrifugal 10min of 1000r/min, get precipitation after the centrifugal 25min of 4000r/min, precipitation ultra-pure water is resuspended.Low speed like this, centrifugal 4 times repeatedly at a high speed, to last ultracentrifugal supernatant achromaticity and clarification, namely obtains purer polyhedral body.By resuspended for ultra-pure water cold for gained polyhedral body 1.5mL, add 200 μ l alkali solution liquid process 15min.Then get supernatant after regulating the centrifugal 5min of pH to 7,5000r/min with the glacial acetic acid of 0.5mol/L, 6000r/min is centrifugal, and 5min gets supernatant, and the centrifugal 5min of 7000r/min, gets supernatant.After at 4 DEG C, ultracentrifugation 1.5h under 28000r/min, precipitates spot 1.5mL ultra-pure water resuspended by gained after abandoning supernatant.
It is the PtCl of 5mM that the bollworm polyhedrosis protein liquid getting 300 μ L alkaline hydrolysis process adds 100 μ L concentration
4solution, fully mixes, in 25 DEG C, and shaking table hatching 24h under 140r/min.Then the NaBH4 solution that concentration that 75 μ L newly prepare is 5mM is dropwise added, limit ultrasonic limit reduction.In 25 DEG C after reduction, under 140r/min, shaking table hatching 6h, namely obtains the nano platinum particle of bollworm polyhedrosis protein load.
Embodiment 3
Former for 0.5g heliothis armigera nuclear polyhedrosis virus powder is dissolved in 60mL ultra-pure water, its dissolving of ultrasonic promotion.Prepared by the PL5124-PALL model ultra-pure water instrument that ultra-pure water used is produced by pure water system Co., Ltd of Vivendi of High Wycombe city of Britain.Get supernatant after the centrifugal 10min of 500r/min, get precipitation after the centrifugal 20min of 4000r/min, precipitation ultra-pure water is resuspended.Low speed like this, centrifugal 3 times repeatedly at a high speed, to last ultracentrifugal supernatant achromaticity and clarification, namely obtains purer polyhedral body.By resuspended for ultra-pure water cold for gained polyhedral body 2mL, add 200 μ l alkali solution liquid process 25min.Then get supernatant after regulating the centrifugal 5min of pH to 7,5000r/min with the glacial acetic acid of 0.5mol/L, 6000r/min is centrifugal, and 5min gets supernatant, and the centrifugal 5min of 7000r/min, gets supernatant.After at 4 DEG C, ultracentrifugation 1h under 28000r/min, precipitates spot 1.5mL ultra-pure water resuspended by gained after abandoning supernatant.Observe its pattern under transmission electron microscope and see Fig. 3.Fig. 3 is the TEM figure of the bollworm polyhedrosis albumen after embodiment 3 purifying and alkaline hydrolysis, and as can be seen from the figure the alkaline hydrolysis time lengthens, and the albumen flakes of the polyhedrosis albumen of gained is more in small, broken bits, and it is 10 ~ 20nm that size is mainly concentrated.
It is the PtCl of 5mM that the bollworm polyhedrosis protein liquid getting 200 μ L alkaline hydrolysis process adds 100 μ L concentration
4solution, fully mixes, in 25 DEG C, and shaking table hatching 20h under 140r/min.Then dropwise adding the concentration that 75 μ L newly prepare is the NaBH of 5mM
4solution, limit ultrasonic limit reduction.In 25 DEG C after reduction, under 140r/min, shaking table hatching 6h, namely obtains the nano platinum particle of bollworm polyhedrosis protein load.Observe its pattern under transmission electron microscope and see Fig. 4.Fig. 4 is the TEM figure of the nano platinum particle of bollworm polyhedrosis load prepared by embodiment 3, from figure can the nano platinum particle decentralization of gained high, size and shape and polyhedrosis albumen basically identical, single nano particle diameter is 4 ~ 5nm, does not have agglomeration.
Embodiment 4
Former for 0.5g heliothis armigera nuclear polyhedrosis virus powder is dissolved in 60mL ultra-pure water, its dissolving of ultrasonic promotion.Prepared by the PL5124-PALL model ultra-pure water instrument that ultra-pure water used is produced by pure water system Co., Ltd of Vivendi of High Wycombe city of Britain.Get supernatant after the centrifugal 5min of 500r/min, get precipitation after the centrifugal 30min of 3000r/min, precipitation ultra-pure water is resuspended.Low speed like this, centrifugal 5 times repeatedly at a high speed, to last ultracentrifugal supernatant achromaticity and clarification, namely obtains purer polyhedral body.By resuspended for ultra-pure water cold for gained polyhedral body 1.5mL, add 150 μ l alkali solution liquid process 10min.Then get supernatant after regulating the centrifugal 5min of pH to 7,5000r/min with the glacial acetic acid of 0.5mol/L, 6000r/min is centrifugal, and 5min gets supernatant, and the centrifugal 5min of 7000r/min, gets supernatant.After at 4 DEG C, ultracentrifugation 1h under 25000r/min, precipitates spot 1mL ultra-pure water resuspended by gained after abandoning supernatant.
It is the PtCl of 1mM that the bollworm polyhedrosis protein liquid getting 100 μ L alkaline hydrolysis process adds 300 μ L concentration
4solution, fully mixes, in 20 DEG C, and shaking table hatching 25h under 130r/min.Then dropwise adding the concentration that 50 μ L newly prepare is the NaBH of 5mM
4solution, limit ultrasonic limit reduction.In 20 DEG C after reduction, under 130r/min, shaking table hatching 6h, namely obtains the nano platinum particle of bollworm polyhedrosis protein load.
Embodiment 5
Former for 1.0g heliothis armigera nuclear polyhedrosis virus powder is dissolved in 100mL ultra-pure water, its dissolving of ultrasonic promotion.Prepared by the PL5124-PALL model ultra-pure water instrument that ultra-pure water used is produced by pure water system Co., Ltd of Vivendi of High Wycombe city of Britain.Get supernatant after the centrifugal 10min of 1200r/min, get precipitation after the centrifugal 20min of 5000r/min, precipitation ultra-pure water is resuspended.Low speed like this, centrifugal 3 times repeatedly at a high speed, to last ultracentrifugal supernatant achromaticity and clarification, namely obtains purer polyhedral body.By resuspended for ultra-pure water cold for gained polyhedral body 2.5mL, add 250 μ l alkali solution liquid process 25min.Then get supernatant after regulating the centrifugal 5min of pH to 8,5000r/min with the glacial acetic acid of 0.5mol/L, 6000r/min is centrifugal, and 5min gets supernatant, and the centrifugal 5min of 7000r/min, gets supernatant.After at 10 DEG C, ultracentrifugation 2h under 30000r/min, precipitates spot 2mL ultra-pure water resuspended by gained after abandoning supernatant.
It is the PtCl of 10mM that the bollworm polyhedrosis protein liquid getting 300 μ L alkaline hydrolysis process adds 100 μ L concentration
4solution, fully mixes, in 30 DEG C, and shaking table hatching 15h under 170r/min.Then dropwise adding the concentration that 100 μ L newly prepare is the NaBH of 10mM
4solution, limit ultrasonic limit reduction.In 30 DEG C after reduction, under 170r/min, shaking table hatching 3h, namely obtains the nano platinum particle of bollworm polyhedrosis protein load.
Claims (2)
1. utilize bollworm polyhedrosis albumen to prepare a method for nano platinum particle, it is characterized in that: described method comprises the steps:
(1) purifying of bollworm polyhedrosis and alkaline hydrolysis:
Former for 0.5 ~ 1.0g heliothis armigera nuclear polyhedrosis virus powder is dissolved in 60 ~ 100mL ultra-pure water, its dissolving of ultrasonic promotion, supernatant is got after the centrifugal 5 ~ 10min of 500 ~ 1200r/min, precipitation is got after the centrifugal 20 ~ 30min of 3000 ~ 5000r/min, precipitation ultra-pure water is resuspended, above-mentioned two centrifugation step are repeated 3 ~ 5 times, to last ultracentrifugal supernatant achromaticity and clarification, namely purer polyhedral body is obtained, gained polyhedral body is resuspended with the ultra-pure water that 1.5 ~ 2.5mL is cold, add 150 ~ 250 μ L alkali solution liquid process 10 ~ 25min, then pH to 7 ~ 8 are regulated with the glacial acetic acid of 0.5mol/L, centrifugal 2 ~ 3 times of 5000 ~ 7000r/min, get supernatant, after at 4 ~ 10 DEG C, ultracentrifugation 1 ~ 2h under 25000 ~ 30000r/min, after abandoning supernatant, gained is precipitated spot 1 ~ 2mL ultra-pure water resuspended,
(2) the noble metal platinum nano particle of bollworm polyhedrosis protein load is prepared:
It is the PtCl of 1 ~ 10mM that the bollworm polyhedrosis protein liquid getting 100 ~ 300 μ L alkaline hydrolysis process adds 100 ~ 300 μ L concentration
4in solution, fully mix, in 20 ~ 30 DEG C, shaking table hatching 15 ~ 25h under 130 ~ 170r/min, then dropwise adding the concentration that 50 ~ 100 μ L newly prepare is the NaBH of 5 ~ 10mM
4solution, limit ultrasonic limit reduction, in 20 ~ 30 DEG C after reduction, under 130 ~ 170r/min, shaking table hatching 3 ~ 6h, namely obtains bollworm polyhedrosis protein load noble metal platinum nano particle.
2. the method utilizing bollworm polyhedrosis albumen to prepare nano platinum particle according to claim 1, it is characterized in that: the alkali solution liquid in described step (1) is 0.3mol/L sodium carbonate, 0.5mol/L sodium chloride and 0.03mol/L disodium ethylene diamine tetra-acetic acid solution.
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| CN104001931B (en) * | 2014-05-12 | 2016-01-06 | 燕山大学 | Bollworm baculoviral nucleocapsid is utilized to prepare the method for precious metal palladium nanometer rods |
| CN104001933B (en) * | 2014-05-12 | 2016-03-02 | 燕山大学 | Bollworm polyhedrosis protein extract is utilized to prepare the method for nanowires of gold |
| CN104668579B (en) * | 2015-02-11 | 2017-04-26 | 燕山大学 | Method for preparing palladium nano wires by use of heliothis armigera nuclear polyhedrin |
| CN104741618B (en) * | 2015-02-13 | 2016-10-05 | 燕山大学 | Utilize the method that bollworm polyhedrosis protein extract prepares Pt nanowires |
| CN114769610B (en) * | 2022-04-02 | 2023-08-11 | 西北工业大学 | Method for preparing gold-palladium nano alloy by utilizing protein assembly |
| CN114854789A (en) * | 2022-05-06 | 2022-08-05 | 苏州大学 | Preparation method of nanocrystal-coated CyHv-2 nucleic acid subunit vaccine |
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