CN105441489A - Method for synthesizing selenium and bismuth selenide nano material by using lysinibacillus sp. - Google Patents
Method for synthesizing selenium and bismuth selenide nano material by using lysinibacillus sp. Download PDFInfo
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- sodium selenite
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- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 title claims abstract description 63
- 239000011669 selenium Substances 0.000 title claims abstract description 55
- 229910052711 selenium Inorganic materials 0.000 title claims abstract description 55
- 239000002086 nanomaterial Substances 0.000 title claims abstract description 38
- FBGGJHZVZAAUKJ-UHFFFAOYSA-N bismuth selenide Chemical compound [Se-2].[Se-2].[Se-2].[Bi+3].[Bi+3] FBGGJHZVZAAUKJ-UHFFFAOYSA-N 0.000 title claims abstract description 25
- 238000000034 method Methods 0.000 title claims abstract description 22
- 230000002194 synthesizing effect Effects 0.000 title abstract description 4
- 241001472591 Lysinibacillus sp. Species 0.000 title 1
- 229940091258 selenium supplement Drugs 0.000 claims abstract description 54
- BVTBRVFYZUCAKH-UHFFFAOYSA-L disodium selenite Chemical compound [Na+].[Na+].[O-][Se]([O-])=O BVTBRVFYZUCAKH-UHFFFAOYSA-L 0.000 claims abstract description 33
- 229960001471 sodium selenite Drugs 0.000 claims abstract description 33
- 239000011781 sodium selenite Substances 0.000 claims abstract description 33
- 235000015921 sodium selenite Nutrition 0.000 claims abstract description 33
- 230000001954 sterilising effect Effects 0.000 claims abstract description 5
- 239000007788 liquid Substances 0.000 claims description 29
- 235000015097 nutrients Nutrition 0.000 claims description 14
- 239000006228 supernatant Substances 0.000 claims description 13
- PPNKDDZCLDMRHS-UHFFFAOYSA-N dinitrooxybismuthanyl nitrate Chemical compound [Bi+3].[O-][N+]([O-])=O.[O-][N+]([O-])=O.[O-][N+]([O-])=O PPNKDDZCLDMRHS-UHFFFAOYSA-N 0.000 claims description 12
- 241000894006 Bacteria Species 0.000 claims description 11
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 8
- XUYPXLNMDZIRQH-LURJTMIESA-N N-acetyl-L-methionine Chemical compound CSCC[C@@H](C(O)=O)NC(C)=O XUYPXLNMDZIRQH-LURJTMIESA-N 0.000 claims description 8
- 229930182817 methionine Natural products 0.000 claims description 8
- 239000002054 inoculum Substances 0.000 claims description 7
- 239000000047 product Substances 0.000 claims description 7
- 238000001035 drying Methods 0.000 claims description 5
- 230000008569 process Effects 0.000 claims description 5
- 239000002244 precipitate Substances 0.000 claims description 4
- 230000015572 biosynthetic process Effects 0.000 abstract description 12
- 239000000243 solution Substances 0.000 abstract description 10
- 238000006243 chemical reaction Methods 0.000 abstract description 7
- 238000012258 culturing Methods 0.000 abstract description 4
- 229910001385 heavy metal Inorganic materials 0.000 abstract description 3
- 238000001179 sorption measurement Methods 0.000 abstract description 3
- 239000001963 growth medium Substances 0.000 abstract 2
- 238000004659 sterilization and disinfection Methods 0.000 abstract 2
- 241001032013 Lysinibacillus sp. ZYM-1 Species 0.000 abstract 1
- 239000007864 aqueous solution Substances 0.000 abstract 1
- RXPAJWPEYBDXOG-UHFFFAOYSA-N hydron;methyl 4-methoxypyridine-2-carboxylate;chloride Chemical compound Cl.COC(=O)C1=CC(OC)=CC=N1 RXPAJWPEYBDXOG-UHFFFAOYSA-N 0.000 abstract 1
- 239000002077 nanosphere Substances 0.000 abstract 1
- 229940082569 selenite Drugs 0.000 description 16
- 239000002105 nanoparticle Substances 0.000 description 13
- 238000003786 synthesis reaction Methods 0.000 description 11
- MCAHWIHFGHIESP-UHFFFAOYSA-L selenite(2-) Chemical compound [O-][Se]([O-])=O MCAHWIHFGHIESP-UHFFFAOYSA-L 0.000 description 8
- 230000001580 bacterial effect Effects 0.000 description 7
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 244000005700 microbiome Species 0.000 description 6
- 239000011734 sodium Substances 0.000 description 6
- 239000007787 solid Substances 0.000 description 6
- 239000002070 nanowire Substances 0.000 description 5
- 239000000843 powder Substances 0.000 description 5
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 4
- 239000001888 Peptone Substances 0.000 description 4
- 108010080698 Peptones Proteins 0.000 description 4
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 4
- DPDMMXDBJGCCQC-UHFFFAOYSA-N [Na].[Cl] Chemical compound [Na].[Cl] DPDMMXDBJGCCQC-UHFFFAOYSA-N 0.000 description 4
- 238000002835 absorbance Methods 0.000 description 4
- 238000009630 liquid culture Methods 0.000 description 4
- 235000019319 peptone Nutrition 0.000 description 4
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 3
- 230000005540 biological transmission Effects 0.000 description 3
- 239000011248 coating agent Substances 0.000 description 3
- 238000000576 coating method Methods 0.000 description 3
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- 238000000870 ultraviolet spectroscopy Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 241001464944 Thauera selenatis Species 0.000 description 2
- 239000007795 chemical reaction product Substances 0.000 description 2
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- QGLWBTPVKHMVHM-KTKRTIGZSA-N (z)-octadec-9-en-1-amine Chemical compound CCCCCCCC\C=C/CCCCCCCCN QGLWBTPVKHMVHM-KTKRTIGZSA-N 0.000 description 1
- PMYDPQQPEAYXKD-UHFFFAOYSA-N 3-hydroxy-n-naphthalen-2-ylnaphthalene-2-carboxamide Chemical compound C1=CC=CC2=CC(NC(=O)C3=CC4=CC=CC=C4C=C3O)=CC=C21 PMYDPQQPEAYXKD-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 239000012901 Milli-Q water Substances 0.000 description 1
- 229940096437 Protein S Drugs 0.000 description 1
- 102000029301 Protein S Human genes 0.000 description 1
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- 241001459308 Pseudomonas alcaliphila Species 0.000 description 1
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- 229960001881 sodium selenate Drugs 0.000 description 1
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to a method for synthesizing selenium and a bismuth selenide nano material by using lysinibacillus sp.ZYM-1. The method comprises the following steps: adding 0.5-25mM of sodium selenite into an LB culture medium subjected to high-temperature sterilization at 121 DEG C, inoculating ZYM-1, and culturing for 24-48h under the condition that the temperature is 25-40 DEG C, the pH value is 5-9 and the speed is 50-300rpm to obtain a red electroselenium nano material; and adding 0.5-25mM of sodium selenite and 0.1-5mM of bismuth nitrate solution into the LB culture medium subjected to high-temperature sterilization at 121 DEG C, inoculating ZYM-1, and culturing for 24-48h under the condition that the temperature is 25-40 DEG C, the pH value is 5-9 and the speed is 50-300rpm to obtain a black bismuth selenide nano material. The reaction is carried out in aqueous solution without severe conditions of high temperature, high pressure and the like, so that cultivation can be enlarged easily. The prepared selenium nano spheres can be applied to adsorption removal of heavy metals, and biosynthesis of the prepared bismuth selenide nano material is not reported at present.
Description
Technical field
The invention belongs to environment field of nano biotechnology, relate to a kind of method utilizing Methionin genus bacillus to synthesize selenium and bismuth selenide nano material.
Background technology
Selenium belongs to the 6th main group element, be the necessary trace element of a kind of biological existence, but the selenium of high density has bio-toxicity and can cause environmental pollution.The selenium of occurring in nature mainly exists with four kinds of valence states (-2,0 ,+4 and+6).Wherein, selenite (SeO
3 2-) soluble in water, compared with other form, there is stronger bio-toxicity.Investigator finds that selenite can be reduced by detoxification by the multiple-microorganism be widespread in nature, and generates toxicity less, the red elemental selenium nano particle that water solvent is poor.WangGJ etc. all report specified microorganisms at document (BMCMicrobiology.2014.14.204-217), LampisS etc. at document (Microbialcellfactories.2014.13.35-39) and OremlandRS etc. in document (Exremophiles.2009.13.695-705) can be reduced to elemental selenium nano particle by Sodium Selenite.But for common microorganism, Sodium Selenite belongs to a kind of toxicant, a small amount of Sodium Selenite may will cause the suppression completely of microorganism growth.
Selenium nano material has excellent unilateral conduction and good photoelectric characteristic, has been widely used in rectifier and photronic manufacture, and its photoelectric characteristic is also for making toner cartridge in Xeroxing.Selenium nanowires is prepared for reducing sodium selenate as AbdelouasA etc. reports pigment C3 in document (Chemistryofmaterials.2000.12.1510-1512).XiaYN etc. report and prepare tripartite's selenium nanowires by liquid phase ageing method in document (JournaloftheAmericanChemicalSociety.2000.122.12582-12583).But these methods need to use expensive reductive agent or the presoma of complexity, or need reaction times of growing very much.Therefore, exploitation reaction conditions is gentle, and the green friendly novel process of building-up process has important using value.Because microorganism has detoxification to selenate and selenite, microorganism can be utilized as " biological factory " of synthesis selenium nano material, realize the Green synthesis process of selenium nano material.As DebieuxCM etc. reports under anaerobic culturing bacterium T.selenatis in document (ProceedingsoftheNationalAcademyofSciences.2011.108.13480-13485), T.selenatis can synthesize the selenium nanometer ball of 150-200nm size, and is transported to outside born of the same parents after being assembled in born of the same parents by Protein S efA.Under the people such as HurHG report anaerobic condition in document (Chemosphere.2007.68.1898-1905), Shewanellasp.HN-41 can synthesize selenium nanometer ball, and by the dimethyl sulfoxide (DMSO) of follow-up interpolation different concns, selenium nanometer ball is converted into nano wire.LiDP utilizes Pseudomonasalcaliphila in aerobic situation, synthesize the mixture of selenium nanometer ball and selenium nanometer ball/selenium nanometer rod in document (ColliodsandSurfaceB:Biointerfaces.2011.88.196-201).But utilize pure bioprocess at present, the selenium nanowires not adding any organic solvent synthesis pure phase has no report.
Bismuth selenide belongs to the inorganic semiconductor material of rhombohedron crystallographic system, and energy gap is narrower, and the thermoelectric conversion efficiency under normal temperature is high, and therefore in heat pump, refrigeration, biosensor etc., tool has been widely used.QianYT etc. report in the mixed solvent of oleyl amine and ethylene glycol in document (SolidStateCommunications.2008.147.36-40), are the hollow Bi of Material synthesis with selenium powder and Bismuth trinitrate
2se
3nano spherical shell structure, the diameter of single spheroid is about 600nm, and shell thickness is only 40nm.But, the Bi reported at present
2se
3synthetic method be chemical method or Physical, not yet have biological process to synthesize the report of bismuth selenide.
Retrieval shows about the document of bismuth selenide synthesis aspect and patent results both at home and abroad, before this invention is filed, does not also find the report of the biosynthesizing aspect of bismuth selenide under aerobic condition.
Summary of the invention
The object of the present invention is to provide a kind of method utilizing Methionin genus bacillus Lysinibacillussp.ZYM-1 reduction selenite synthesis selenium nano material and bismuth selenide nano material, selenium nano material and bismuth selenide nano material are in photochemical catalysis, and heavy metal adsorption and thermoelectric conversion field have significant application value.
Methionin genus bacillus Lysinibacillussp.ZYM-1 is that contriver is separated and obtains from the bed mud sample of Panjin City Liaodong Wan coastal waters, Liaoning Province, be preserved in China Microbial Culture Preservation Commission's common micro-organisms center (address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica), preservation date is on October 22nd, 2015, culture presevation is numbered CGMCC1.15346, and the Genbank number of logging in of its 16srRNA gene order is KT263530.
Technical scheme of the present invention is:
Methionin genus bacillus Lysinibacillussp.ZYM-1, is characterized in that: Methionin genus bacillus ZYM-1 has two kinds of training methods:
A kind of is melted at 4 DEG C by the glycerol stock liquid of-80 DEG C of bacterial strain ZYM-1 preserved, and is inoculated in 20ml and cultivates containing in the LB liquid nutrient medium of 0.5-25mM (mmole) Sodium Selenite; Described LB liquid culture based formulas is: sodium-chlor 10g/L, peptone 10g/L, yeast powder 5g/L; LB liquid nutrient medium all needs to carry out 121 DEG C of sterilising treatment 20min before using.
Another kind adopts LB solid medium, the ZYM-1 bacterium liquid flat board coating after above-mentioned cultivation cultivated; Described LB solid medium is in LB liquid nutrient medium, add the agar powder that massfraction is 2%.
A kind of ZYM-1 of utilization synthesizes the method for selenium nano material: ZYM-1 is inoculated in 20ml and cultivates 8-12 hour containing in the LB liquid nutrient medium of 0.5-25mM Sodium Selenite; ZYM-1 bacterium liquid after cultivation is transferred in new 100mlLB liquid nutrient medium with the inoculum size of 1%-5%, add the Sodium Selenite of 0.5-25mM, 25-40 DEG C, pH5-9,24-48h is cultivated under 50-300rpm, 20min is processed at ZYM-1 bacterium liquid after cultivation is placed in 121 DEG C, nanometer selenium in born of the same parents is discharged from thalline, 2000-3000rpm is centrifugal, the red supernatant of separating thallus fragment and wrapping biological molecule, red supernatant 12000-20000rpm is centrifugal, get centrifugal product, after drying, obtain red selenium nano material.
A kind of ZYM-1 of utilization synthesizes the method for bismuth selenide nano material: ZYM-1 is inoculated in 20ml and cultivates 8-12 hour containing in the LB liquid nutrient medium of 0.5-25mM Sodium Selenite; ZYM-1 bacterium liquid after cultivation is transferred in new 100mlLB liquid nutrient medium with the inoculum size of 1%-5%, add the Sodium Selenite of 0.5-25mM and the Bismuth trinitrate of 0.1-5mM simultaneously, 25-40 DEG C, pH5-9, cultivate 24-48h under 50-300rpm, subsequently 2000-3000rpm collected by centrifugation product, remove and precipitate to obtain black supernatant liquor, black supernatant liquor is used the centrifugal acquisition throw out of 12000-20000rpm again, black bismuth selenide nano material after drying precipitate, can be obtained.Transmission electron microscope and atomic force microscope display bismuth selenide are three-dimensional ball shape structure, and size is about between 20-40nm.
The invention has the beneficial effects as follows: the selenite larger containing selenium waste water toxic is synthesized selenium nano material and bismuth selenide nano material in the condition of gentleness by the present invention, method environment is green friendly, reaction is carried out in aqueous, without the need to severe condition such as High Temperature High Pressure, simple to operate, be easy to enlarged culturing.The appearance of nano material of preparation is controlled, and selenium nanometer ball may be used for the Adsorption of heavy metal, and bismuth selenide nano material has not yet to see biosynthetic report.
Accompanying drawing explanation
Fig. 1 is the digital photograph of ZYM-1 reduction selenite synthesis selenium nano particle under dull and stereotyped coating culture condition.
Fig. 2 is the variation tendency of selenium also commercial weight under different concentration of sodium selenite.
Fig. 3 is the variation tendency of selenium also commercial weight under different pH.
Fig. 4 is the ultraviolet full wavelength scanner figure synthesizing selenium nanoparticle system under different concentration of sodium selenite.
Fig. 5 is the topography scan Electronic Speculum figure synthesizing selenium nano particle under different concentration of sodium selenite;
A () is for generating the linear selenium nano material of nanometer after the reduction of 1mM Sodium Selenite;
B () is for generating cubes or spherical selenium nano material after the reduction of 2.5mM Sodium Selenite;
C () is for generating the selenium nano material of nanometer spherical after the reduction of 5mM Sodium Selenite;
D () is for generating irregular selenium nano material after the reduction of 25mM Sodium Selenite.
Fig. 6 is the representative transmission electron microscope picture that ZYM-1 synthesizes bismuth selenide.
Fig. 7 is the atomic force microscope collection of illustrative plates that ZYM-1 synthesizes bismuth selenide.
Embodiment
Embodiment 1: bacterial strain ZYM-1 reduction selenite synthesis selenium nano particle under dull and stereotyped coating culture condition
Reaction system: after the ZYM-1 bacterium liquid dilution different multiples after cultivate 8-12 hour in the LB liquid nutrient medium of 20ml containing 0.5-25mM (mmole) Sodium Selenite, with being coated with on LB solid medium that rod is coated on containing 1mM Sodium Selenite uniformly.36h is cultivated in 30 DEG C of constant incubators.The color from light yellow of bacterium colony becomes redness, discloses the generation of reduzate selenium nano particle, and the dull and stereotyped photo of cultivation as shown in Figure 1.
Embodiment 2: under different concentration of sodium selenite, bacterial strain ZYM-1 is to the reducing degree of selenite
In the LB liquid medium of 20mL, add the Sodium Selenite of 0.5,1,2.5,5,10,25mM respectively, ZYM-1 bacterium liquid inoculum size is 1%, and culture temperature is 30 DEG C, and pH is that 5.5,100rpm cultivates 48h; Described LB liquid culture based component is: sodium-chlor 10g/L, peptone 10g/L, yeast powder 5g/L.The elemental selenium nano particle Na generated
2s is oxidized, and calculate selenium simple substance content, concrete grammar is as follows:
1. prepare the Na of 1M
2s solution, adds appropriate NaOH solid and suppresses Na
2certainly the hydrolysis of S.The NaCl solution of configuration 1M.In 10ml test tube, add the commercialization selenium powder of different amount respectively, add 5mLNa
2s solution fully vibrates dissolving, makes the concentration of selenium in reference liquid be respectively 0.5,1,2,3,4,5mM.Utilize the absorbance (OD of ultraviolet-visible spectrophotometer bioassay standard solution under 500nm
500nm), drawing standard curve.Wherein ordinate zou is absorbance OD
500nm, X-coordinate is corresponding selenium concentration of standard solution.The relation of both correspondences utilizes linear fit equation to obtain.
2. the sample containing selenium nano particle in thalline and born of the same parents is passed through the centrifugal 20min of 12000rpm, be separated with supernatant liquor.Utilize 1MNaCl washing precipitation, remove selenite radical remaining in solution, subsequently, by the Na of residual solids 5mL1M
2s fully vibrates dissolving, utilizes ultraviolet-visible spectrophotometer under 500nm, measure the absorbance of solution.The typical curve linear fit result utilizing step 1 to obtain, can from the OD of sample actual measurement
500nmcalculate the content of corresponding selenium element.As measured absorbancy OD
500nmin the absorbancy exceeding typical curve in limited time, need sample to dilute, ensure that the absorbancy measured is in the scope that typical curve comprises.Experimental result as shown in Figure 2.When the content of initial selenite radical is 0.5,1,2.5 and 5mM time, the transformation efficiency of bacterial strain ZYM-1 in 48h all can reach 100%, and when initial selenite radical concentration rises to 10mM, transformation efficiency reduces to 53%, when initial selenite radical concentration is 25mM, transformation efficiency is 31.7%.
Embodiment 3: different pH is on the impact of bacterial strain reduction selenite radical.
Utilize HCl or NaOH to be 4,5,6,7,8,9,10,11 by the pH regulator of LB liquid medium respectively, and ensure that culture system cumulative volume is 20mL.Bacterium liquid ZYM-1 inoculum size is 1%, and selenite radical concentration is 5mM, and culture temperature is 30 DEG C, and 200rpm cultivates 36h; Described LB liquid culture based component is: sodium-chlor 10g/L, peptone 10g/L, yeast powder 5g/L.Utilize Na
2the elemental selenium nano particle that S oxidation generates, concrete measuring method is as follows:
By the sample containing selenium nano particle in thalline and born of the same parents by the centrifugal 10min of 10000rpm, be separated with supernatant liquor.Utilize 1MNaCl solution to rinse 2 times, remove selenite radical remaining in solution, subsequently, by the Na of residual solids 5mL1M
2s fully vibrates dissolving, utilizes ultraviolet-visible spectrophotometer under 500nm, measure the absorbance of solution.The amount of the elemental selenium in solution is calculated according to typical curve.Experimental result as shown in Figure 3.Bacterial strain ZYM-1 effectively can reduce to the selenite radical of 5mM in the scope that pH is 5-9, and reduction ratio is respectively 88%, 98%, 99%, 95% and 93%.
Embodiment 4: different initial concentration of sodium selenite is on the impact of synthesis selenium appearance of nano material.
Embodiment 2 is reacted the thalline comprising selenium nano particle of acquisition by 121 DEG C of damp and hot pyroprocessing 20min, mixture after process is centrifugal 5min under the condition of 3000rpm, the red supernatant of separating thallus fragment and wrapping biological molecule, get supernatant liquor centrifugal 20min under 12000rpm subsequently, obtain centrifugal product, product is the selenium nano material of acquisition after carrying out washing drying.By resuspended in distilled water for the selenium nano particle obtained, carry out visible region full wavelength scanner, result as shown in Figure 4.Fig. 4 shows the absorption spectrogram of 0.5-25mM Sodium Selenite system reaction product in 450-800nm interval.As can be seen from the figure the product that the reaction of 1mM and 5mM is complete has maximum absorption peak at 580nm and 592nm place respectively.The reaction product of other concentration does not have obvious absorption peak in described region.
To comprising 1mM, the reduzate of the Sodium Selenite of 2.5mM, 5mM, 25mM has carried out scanning electron microscope sign, and result as shown in Figure 5.As can be seen from the figure obvious pattern evolutionary process.What Fig. 5 a showed is the selenium nano material generated after the Sodium Selenite reduction of 1mM, and its pattern is presented as homogeneous nano wire form, and its length is 5-10 μm, and width is 50-100nm.When concentration continues to be increased to 2.5mM, the pattern of selenium nano material is presented as that cubes is main, and part is the state of ball-type, as shown in Figure 5 b.When concentration is increased to 5mM, selenium nano material develops into nanometer ball substantially completely, diameter at about 300nm, as shown in Figure 5 c.When concentration continues to be increased to 25mM, selenium nano material develops into irregular pattern, as fig 5d.
Embodiment 5: bacterial strain ZYM-1 is used for the synthesis of bismuth selenide nano material
The Sodium Selenite of 0.5,1,2.5,5,10,25mM and the Bismuth trinitrate of corresponding Sodium Selenite substance withdrawl syndrome 1/10 is added respectively in the LB liquid medium of 20mL.Sodium Selenite system as 0.5mM adds the Bismuth trinitrate of 0.05mM, and the Sodium Selenite system of 1mM adds the Bismuth trinitrate of 0.1mM, and other system is similar.Inoculum size is 1%, and culture temperature is 30 DEG C, and pH is that 5.5,300rpm cultivates 24h; Described LB liquid culture based component is: sodium-chlor 10g/L, peptone 10g/L, yeast powder 5g/L.Supernatant liquor after 3000rpm centrifugal 5min collection is centrifugal, then obtain product with the centrifugal 20min of 12000rpm, utilize milli-Q water 3 times, in 80 DEG C of baking ovens, bismuth selenide nano material can be obtained after dry 6h.Representational bismuth selenide transmission electron microscope and atomic force microscope figure are as shown in Fig. 6, Fig. 7.As can be seen from the figure, the bismuth selenide of synthesis is mainly ball-type, and size is about 20nm.
The above; be only this patent preferably embodiment; but protection scope of the present invention is not limited in this; even if those skilled in the art are familiar with in the scope that the present invention discloses any; be equal to according to technological method of the present invention and inventive concept thereof and replace or change, all should be encompassed within protection scope of the present invention.
Claims (2)
1. utilize Methionin genus bacillus ZYM-1 to synthesize a method for selenium nano material, it is characterized in that: ZYM-1 is inoculated in 20ml containing 8-12 hour in the LB liquid nutrient medium of 0.5-25mM Sodium Selenite; ZYM-1 bacterium liquid after cultivation is transferred in 100mlLB liquid nutrient medium with the inoculum size of 1%-5%, adds the Sodium Selenite of 0.5-25mM, cultivates 24-48h under 25-40 DEG C, pH5-9,50-300rpm; ZYM-1 bacterium liquid 121 DEG C process 20min, 2000-3000rpm is centrifugal obtains red supernatant, and red supernatant 12000-20000rpm is centrifugal, obtains selenium nano material after centrifugal product drying; Described LB liquid nutrient medium needs 121 DEG C of sterilising treatment 20 minutes before using.
2. utilize Methionin genus bacillus ZYM-1 to synthesize a method for bismuth selenide nano material, it is characterized in that: ZYM-1 is inoculated in 20ml containing 8-12 hour in the LB liquid nutrient medium of 0.5-25mM Sodium Selenite; ZYM-1 bacterium liquid after cultivating is transferred in 100mlLB liquid nutrient medium with the inoculum size of 1%-5%, adds the Sodium Selenite of 0.5-25mM and the Bismuth trinitrate of 0.1-5mM, under 25-40 DEG C, pH5-9,50-300rpm, cultivate 24-48h; 2000-3000rpm is centrifugal, removes and precipitates to obtain black supernatant liquor, by centrifugal for black supernatant liquor 12000-20000rpm acquisition throw out, obtains bismuth selenide nano material after drying precipitate; Described LB liquid nutrient medium needs 121 DEG C of sterilising treatment 20 minutes before using.
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