CN110257311B - Microorganism for preparing broccoli-shaped antimony sulfide and application thereof - Google Patents

Microorganism for preparing broccoli-shaped antimony sulfide and application thereof Download PDF

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CN110257311B
CN110257311B CN201910743357.1A CN201910743357A CN110257311B CN 110257311 B CN110257311 B CN 110257311B CN 201910743357 A CN201910743357 A CN 201910743357A CN 110257311 B CN110257311 B CN 110257311B
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antimony sulfide
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salinity
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CN110257311A (en
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张海坤
解静仪
孙延瑜
胡晓珂
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Yantai Institute of Coastal Zone Research of CAS
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Abstract

The invention belongs to the technical field of water treatment in environmental engineering, and particularly relates to a microorganism for preparing broccoli-shaped antimony sulfide and application thereof. The microorganism related to the invention is marine halomonasHalomonassp.x3, deposited in the chinese culture collection, address: wuhan university, the preservation time is 2019, 5 months and 30 days, and the preservation number is: CCTCC M2019409. The strain is indigenous microorganisms screened from marine sediments, and can prepare nano antimony sulfide by using potassium pyroantimonate and sodium thiosulfate or sodium sulfite as precursors. The nano antimony sulfide obtained by the microorganism is broccoli-shaped, and can be used as a photocatalyst for accelerating the photodegradation of azo dye compounds. The preparation of the antimony sulfide mostly adopts a chemical method at present, and the invention prepares the antimony sulfide with controllable appearance by utilizing microorganisms for the first time, thereby having stronger innovation.

Description

Microorganism for preparing broccoli-shaped antimony sulfide and application thereof
Technical Field
The invention relates to the technical field of water treatment in environmental engineering, in particular to a microorganism for preparing broccoli-shaped antimony sulfide and application thereof.
Background
In recent years, binary metal chalcogenides have attracted much attention from researchers because of their excellent photoelectric properties. The band gaps of the metal chalcogenide are narrow, and the narrow band gaps can enable the metal chalcogenide to widely absorb visible light and near infrared light in sunlight, so that the metal chalcogenide has a good application prospect in photovoltaic devices and thermoelectric devices. Wherein antimony trisulfide (Sb)2S3) The band gap of the solar cell is about 1.6eV, is close to the optimal theoretical band gap value of a single-junction solar cell, and the solar cell and a semiconductor photoelectric/photovoltaic device are widely applied. In addition, Sb2S3The nano material also has high specific surface area and can be used in a photocatalytic reactor based on visible light. Thus, study of Sb2S3Efficient and controllable synthesis of nanomaterials is in the fieldThe direction of emphasis of.
Sb reported at present2S3The synthesis methods are mostly chemical methods, such as hydrothermal method, wet chemical method, microwave method, reflux method, colloid reaction method, low-temperature solvothermal method and the like. The methods can synthesize Sb with different shapes under certain conditions2S3Such as rod, dumbbell, chrysanthemum, sea urchin, dandelion, etc. However, some of these methods are relatively complex, some have severe condition control, some require addition of a large amount of toxic chemicals, and have certain limitations in practical application processes. Therefore, there is a great need to develop Sb which is environment-friendly, low in cost and easy to operate2S3The method of (1).
The biological synthesis nano material has the characteristics of low cost, environmental friendliness and the like, and can be used for pertinently developing Sb2S3The microbe synthesis technology of nanometer material. It has been reported that microorganisms can be used to synthesize various metal chalcogenides such as CuS, ZnS, FeS, etc. However, since Sb (V) compounds are toxic and difficult to reduce, no microorganism has been used for synthesizing Sb2S3And (4) reporting of nano materials.
Disclosure of Invention
The invention aims to provide a microorganism for preparing broccoli-shaped antimony sulfide and application thereof.
In order to achieve the purpose, the invention adopts the technical scheme that:
a microorganism for the preparation of broccoli-like antimony sulfide, the microorganism being the marine bacterium Halomonas sp.x3, deposited at the chinese collection of cultures, address: china, Wuhan and Wuhan university, the preservation time is 2019, 5 months and 30 days, and the preservation number is: CCTCC M2019409.
The strain grows under the conditions of 10-50 ℃, 1-7% of salinity and 4-10 of pH.
The application of a microorganism in preparing broccoli-shaped antimony sulfide.
Inoculating 0.1-1g/L dry weight of marine bacteria into a culture medium containing Sb (V) and S2O3 2-(orSO3 2-) In the inorganic salt culture medium, 0.5-2g/L carbon source is added for reduction reaction at 10-50 ℃, 1-7% of salinity and pH of 4-10, and the broccoli-shaped Sb is obtained after hot water bath treatment2S3(ii) a The carbon source is sodium lactate, glucose, sodium formate or sucrose.
Further, it is said that
1) The marine bacteria are cultured in a medium containing 0.01-2mM Sb (V) and 0.5-2mM S2O3 2-(or SO)3 2-) The inorganic salt culture medium is a mixture which is subjected to light-shielding reaction for 1-10 days at the temperature of 10-50 ℃, the salinity of 1-7 percent and the pH value of 4-10;
2) centrifuging the mixture obtained in the step (1), placing the obtained precipitate in deionized water for ultrasonic treatment, after centrifugation, resuspending the precipitate with 0.2-5% of anionic surfactant (sodium dodecyl sulfate), treating for 1-45min in a hot water bath at 70-100 ℃, then centrifuging and freeze-drying the precipitate to obtain powdery particles, namely the biologically prepared broccoli-shaped Sb2S3。
The temperature is 20-40 ℃, the salinity is 1-3% and the pH is 6-8.
Broccoli-shaped Sb2S3The preparation of (1) inoculating the marine bacteria with dry weight of 0.1-1g/L into a culture medium containing Sb (V) and S2O3 2-(or SO)3 2-) In the inorganic salt culture medium, 0.5-2g/L carbon source is added for reduction reaction at 10-50 ℃, 1-7% of salinity and pH of 4-10, and the broccoli-shaped Sb is obtained after hot water bath treatment2S3(ii) a The carbon source is sodium lactate, glucose, sodium formate or sucrose.
In a further aspect of the present invention,
1) the marine bacterium according to claim 1, wherein the marine bacterium contains 0.01-2mM Sb (V) and 0.5-2mM S2O3 2-(or SO)3 2-) Adding 0.5-2g/L carbon source into the inorganic salt culture medium at 10-50 deg.C, 1-7% salinity and pH of 4-10, and reacting in dark for 1-10 days to obtain a mixture;
2) centrifuging the mixture obtained in the step (1), placing the obtained precipitate in deionized water for ultrasonic treatment, centrifuging, and using the precipitateResuspending 0.2-5% cationic surfactant (sodium dodecyl sulfate), treating with 70-100 deg.C hot water bath for 1-45min, centrifuging, and freeze drying the precipitate to obtain powdery granule, i.e. biological prepared cauliflower-like Sb2S3。
Preparation of broccoli-shaped Sb2S3The application of the antimony sulfide in degrading azo compounds in the dye wastewater by light.
The nano Sb with the ultrasonic dispersion concentration of 1-100mg/L is added into water2S3Adding the dispersion liquid into the dye wastewater to be treated, and carrying out photocatalytic degradation under the irradiation of a long-arc xenon lamp (500W) at room temperature so as to realize the degradation of azo compounds in the dye wastewater; wherein, each liter of wastewater contains nano Sb2S3The amount of (A) is 1-100 mg.
The invention has the advantages that:
the method screens indigenous microorganisms from marine sediments, and can prepare nano antimony sulfide by using potassium pyroantimonate and sodium thiosulfate or sodium sulfite as precursors. The nano antimony sulfide obtained by the microorganism is in a broccoli shape, is synthesized by a microorganism method with controllable appearance, has low technical cost and is environment-friendly, and has more industrial value compared with a chemical method. The nanometer antimony sulfide obtained by the microorganism can be used as a photocatalyst for accelerating the photodegradation of azo dyes compounds.
Drawings
Figure 1 is a graph of the color change of Halomonas sp.x3, provided in example 1 of the present invention, after antimony sulfide was produced within 5 d. Wherein, 1 bacterium + Sb (V) + SO3 2-(ii) a 2 bacteria + Sb (V) + S2O3 2-(ii) a 3 bacteria + Sb (V).
FIG. 2 is the X-ray photoelectron spectrum of the biological nanometer antimony sulfide in example 2. Wherein (a) is a peak separation diagram of different valences of antimony and oxygen; (b) is a diagram of the peaks of different valence states of sulfur.
FIGS. 3a-b are the electron scanning electron micrographs of biological nano antimony sulfide mentioned in example 2 of the present invention (a)1 μm; (b)500 nm; FIGS. 3c-d Electron Transmission Electron micrographs (c) of biological Nano antimony sulfide mentioned in example 2 of the present invention at 50 nm; (d)100 nm.
FIG. 4 is a scanning electron microscopy (EDS) spectrum of the biological nano antimony sulfide mentioned in example 3 of the present invention.
FIG. 5 is a graph showing the catalytic effect of bio-nano antimony sulfide according to example 4 of the present invention, wherein the bio-nano antimony sulfide is subjected to ultrasonic treatment for 1 hour before use.
Detailed Description
The following non-limiting examples are presented to enable those of ordinary skill in the art to more fully understand the present invention and are not intended to limit the invention in any way.
The method takes marine bottom sediment in the middle of a Bohai sea channel as a bacterial source, screens a marine indigenous bacterium, is named as X3, can be expanded and propagated by using improved LB and marine 2216E and other classical culture media, and can also grow in an inorganic salt culture medium by using various carbon sources (such as sodium lactate, glucose, sodium formate and sucrose) under the conditions of 10-50 ℃, 1-7% salinity and 4-10 pH. The improved LB culture medium is 10g of peptone, 5g of yeast extract, 20g of NaCl and the balance of deionized water per liter; 5g of peptone, 1g of yeast extract, 0.01g of phosphoric acid and ferric iron and the balance of aged seawater are added into each liter of the ocean 2216E culture medium; the inorganic salt culture medium is 1g NH per liter4Cl,0.8g Na2HPO4,0.2g KH2PO4,0.2g MgCl2·7H2O,0.1g CaCl2·2H2O,20g NaCl, and the balance of deionized water.
X3 is gram-negative bacteria, bacterial colony is light brown, irregular round, smooth in surface and free of rough edges, the bacteria are found to be single-grown and rod-shaped through identification of an electronic scanning electron microscope, 16s rRNA shows that the similarity with the Boliviana Halomonas (Halomonas boliviansis LC1) is highest through comparison (96.85 percent), so the bacterial colony is named as Halomonas sp.X3, the bacterial colony is preserved in China center for type culture collection, the address is eight Lopa mountains in Wuhan city, Gao, Hubei, the preservation time is 6 months 19 in 2019, and the preservation number is: CCTCC M2019409. X3 was identified as producing broccoli-like antimony sulfide, and subsequent examples were based on the above studies.
X3 16 srna sequence:
CGAGCGGTAACAGATCCAGCTTGCTGGATGCTGACGAGCGGCGGACGGGTGAGTAATGCATAGGAATCTGCCCGNTAGTGGGGGATAACCTGGGGAAACCCAGGCTAATACCGCATACGTCCTACGGGAGAAAGGGGGCTTCGGCTCCCGCTATCGGATGAGCCTATGTCGGATTAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCCACGATCCGTAGCTGGTCTGAGAGGATGATCAGCCACATCGGGACTGAGACACGGCCCGAACTCCTACGGGAGGCAGCAGTGGGGAATATTGGACAATGGGGGGAACCCTGATCCAGCCATGCCGCGTGTGTGAAGAAGGCCCTCGGGTTGTAAAGCACTTTCAGCGAGGAAGAACGCCTAGCGGTTAATACCCGCTAGGAAAGACATCACTCGCAGAAGAAGCACCGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGGAGGGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAGCGCGCGTAGGTGGCTTGATAAGCCGGTTGTGAAAGCCCCGGGCTCAACCTGGGAACGGCATCCGGAACTGTCAAGCTAGAGTGCAGGAGAGGAAGGTAGAATTCCCGGTGTAGCGGTGAAATGCGTAGAGATCGGGAGGAATACCAGTGGCGAAGGCGGCCTTCTGGACTGACACTGACACTGAGGTGCGAAAGCGTGGGTAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGTCGACCAGCCGTTGGGTGCCTAGCGCACTTTGTGGCGAAGTTAACGCGATAAGTCGACCGCCTGGGGAGTACGGCCGCAAGGTTAAAACTCAAATGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGATGCAACGCGAAGAACCTTACCTACTCTTGACATCCTGCGAATTNGGTAGAGATACCTTAGTGCCTTCGGGAACGCAGAGACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTTGTGAAATGTTGGGTTAAGTCCCGTAACGAGCGCAACCCTTGTCCTTATTTGCCAGCGCGTAATGGCGGGAACTCTAAGGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGACGACGTCAAGTCATCATGGCCCTTACGAGTAGGGCTACACACGTGCTACAATGGTCGGTACAAAGGGTTGCCAACTCGCGAGAGTGAGCCAATCCCGAAAAGCCGATCTCAGTCCGGATCGGAGTCTGCAACTCGACTCCGTGAAGTCGGAATCGCTAGTAATCGTAGATCAGAATGCTACGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGGAGTGGACTGCACCAGAAGTGGTTAGCCTAACGCAAGAGGGCGATCACCA
example 1
The above-obtained strain X3 (dry weight 0.5g/L), Sb (V) (0.1mM) and 1mM S2O3 2-(or SO)3 2-) Mixing in 100mL inorganic salt culture medium with salinity of 2%, adjusting pH to 7, adding 18mM sodium lactate, exposing to nitrogen for 15min, reacting in 30 deg.C anaerobic incubator in dark place for 5d, and changing color as shown in FIG. 1.
The inorganic salt culture medium is 1g NH per liter4Cl,0.8g Na2HPO4,0.2g KH2PO4,0.2g MgCl2·7H2O,0.1g CaCl2·2H2O,20g NaCl, and the balance of deionized water.
As can be seen from FIG. 1, X3 can turn orange yellow in the presence of antimony and thiosulfate or sulfite, which is verified as Sb2S3Wherein the darker the color, the better the reduction. The results show that in the current reaction system, the reaction system with sulfite added is lighter in color than the reaction system with thiosulfate added at the same time point (see fig. 1), indicating that the thiosulfate effect is more prominent than the sulfite effect.
Example 2
The strains X3(0.5g/L), Sb (V) (0.2mM) and 1mM S obtained above were added2O3 2-Mixing in 100mL inorganic salt culture medium with salinity of 2%, adjusting pH to 7, adding 18mM sodium lactate, exposing to nitrogen for 15min, reacting in 30 deg.C anaerobic incubator in dark place for 5d, centrifuging (10000rpm,5min), collecting precipitate, ultrasonic treating in deionized water for 1h, centrifuging, resuspending the precipitate with 1% anionic surfactant sodium dodecyl sulfate, treating in 90 deg.C hot water bath for 45min, centrifuging again (10000rpm, 10min), and freeze drying to obtain Sb2S3(see FIG. 2).
The inorganic salt culture medium is 1g NH per liter4Cl,0.8g Na2HPO4,0.2g KH2PO4,0.2g MgCl2·7H2O,0.1g CaCl2·2H2O,20g NaCl, and the balance of deionized water.
The product obtained is powder Sb which is verified by X-ray diffraction energy spectrum in figure 22S3Wherein 3d of Sb (V) is shown in FIG. 2a5/2The binding energy site appears at 531eV, and 3d of Sb (III)5/2The binding energy site appeared at 528.2eV, from which it was concluded that most of Sb (V) was reduced to Sb (III). At the same time, from the XPS plot of S (FIG. 2b), S2O3 2-Is substantially reduced to S2-One part of the antimony sulfide is combined with Sb (III) to form antimony sulfide, and the other part of the antimony sulfide is free in the system.
The shape of the resulting material was verified by electron scanning electron microscopy and transmission electron microscopy (see FIGS. 3 a-d). 3a-b, the arrow points to the position to produce the nanometer antimony sulfide material in a broccoli shape. As can be seen from FIGS. 3c-d, the pointed position of the arrow is the generated nano antimony sulfide material, which conforms to the shape obtained by scanning electron microscopy.
Example 3
The strain X3(0.7g/L), Sb (V) (1mM) and 2mM SO obtained above were added3 2-Mixing in 100mL inorganic salt culture medium with salinity of 3%, adjusting pH to 7.2, adding 18mM glucose, exposing to nitrogen for 15min, reacting in 30 deg.C anaerobic incubator in dark place for 7d, centrifuging the obtained mixture (10000rpm,5min), placing the precipitate in deionized water for ultrasonic treatment for 1h, resuspending the precipitate with 1.5% anionic surfactant sodium dodecyl sulfate, treating in 95 deg.C hot water bath for 60min, centrifuging again (10000rpm, 10min), and freeze drying the precipitate to obtain Sb2S3。
The elements contained in the obtained material are verified by an energy spectrometer, and the material is mainly composed of Sb and S elements according to the energy spectrum result, and in addition, the surface has some residual salts (such as Na, Cl and P) and biological elements (such as C, N, O, P) (figure 4).
Example 4
The biological nano antimony sulfide obtained in the example 2 is used for carrying out photodegradation detection on methyl orange in azo dye:
and (3) setting a control group and an experimental group, wherein the experimental group is added with the biological nano antimony sulfide obtained in the example 2 and is used as a catalyst, and the control group is a system without the catalyst: wherein the experimental group reaction system is 50mL of deionized water, 10mg/L of methyl orange and 30mg/L of the biological nano antimony sulfide treated by ultrasonic treatment prepared by the above embodiment, and the comparison system is 50mL of deionized water and 10mg/L of methyl orange. Then, photocatalytic degradation was carried out at room temperature under irradiation of a long-arc xenon lamp (500W) (irradiation distance: about 10cm), 3mL of each of the reaction systems of the above experimental group and control group was sampled, and the change in concentration of methyl orange was monitored at 462nm using a spectrophotometer (see FIG. 5).
Fig. 5 shows that the degradation rate of azo dye in the experimental group containing the material is significantly improved compared with that of the control group, the degradation rate of methyl orange in the blank control system is only about 5% within 3h of reaction time, and the degradation rate of methyl orange in the experimental group containing the material can reach 82%.
Sequence listing
<110> institute of tobacco pipe coastal zone of Chinese academy of sciences
<120> a microorganism for preparing broccoli-like antimony sulfide and use thereof
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1397
<212> DNA
<213> Marine bacterium (Halomonas sp. X3)
<400> 1
cgagcggtaa cagatccagc ttgctggatg ctgacgagcg gcggacgggt gagtaatgca 60
taggaatctg cccgntagtg ggggataacc tggggaaacc caggctaata ccgcatacgt 120
cctacgggag aaagggggct tcggctcccg ctatcggatg agcctatgtc ggattagcta 180
gttggtgagg taacggctca ccaaggccac gatccgtagc tggtctgaga ggatgatcag 240
ccacatcggg actgagacac ggcccgaact cctacgggag gcagcagtgg ggaatattgg 300
acaatggggg gaaccctgat ccagccatgc cgcgtgtgtg aagaaggccc tcgggttgta 360
aagcactttc agcgaggaag aacgcctagc ggttaatacc cgctaggaaa gacatcactc 420
gcagaagaag caccggctaa ctccgtgcca gcagccgcgg taatacggag ggtgcaagcg 480
ttaatcggaa ttactgggcg taaagcgcgc gtaggtggct tgataagccg gttgtgaaag 540
ccccgggctc aacctgggaa cggcatccgg aactgtcaag ctagagtgca ggagaggaag 600
gtagaattcc cggtgtagcg gtgaaatgcg tagagatcgg gaggaatacc agtggcgaag 660
gcggccttct ggactgacac tgacactgag gtgcgaaagc gtgggtagca aacaggatta 720
gataccctgg tagtccacgc cgtaaacgat gtcgaccagc cgttgggtgc ctagcgcact 780
ttgtggcgaa gttaacgcga taagtcgacc gcctggggag tacggccgca aggttaaaac 840
tcaaatgaat tgacgggggc ccgcacaagc ggtggagcat gtggtttaat tcgatgcaac 900
gcgaagaacc ttacctactc ttgacatcct gcgaattngg tagagatacc ttagtgcctt 960
cgggaacgca gagacaggtg ctgcatggct gtcgtcagct cgtgttgtga aatgttgggt 1020
taagtcccgt aacgagcgca acccttgtcc ttatttgcca gcgcgtaatg gcgggaactc 1080
taaggagact gccggtgaca aaccggagga aggtggggac gacgtcaagt catcatggcc 1140
cttacgagta gggctacaca cgtgctacaa tggtcggtac aaagggttgc caactcgcga 1200
gagtgagcca atcccgaaaa gccgatctca gtccggatcg gagtctgcaa ctcgactccg 1260
tgaagtcgga atcgctagta atcgtagatc agaatgctac ggtgaatacg ttcccgggcc 1320
ttgtacacac cgcccgtcac accatgggag tggactgcac cagaagtggt tagcctaacg 1380
caagagggcg atcacca 1397

Claims (6)

1. A microorganism for the production of broccoli-like antimony sulfide, characterized in that: the microorganism is marine bacteriaHalomonassp, X3, as deposited with the chinese type culture collection, address: china, Wuhan and Wuhan university, the preservation time is 2019, 5 months and 30 days, and the preservation number is: CCTCC M2019409.
2. The microorganism according to claim 1 for the preparation of broccoli-like antimony sulfide, wherein: the growth condition of the microorganism is 10-50oC, salinity of 1-7% and pH of 4-10.
3. Use of a microorganism according to claim 1, wherein: the application of the microorganism in preparing broccoli-shaped antimony sulfide.
4. Broccoli-shaped Sb2S3The preparation method is characterized by comprising the following steps: inoculating the microorganism of claim 1 with a dry weight of 0.1-1g/L into the culture medium at 10-50oC. Adding 0.5-2g/L carbon source for reduction reaction under the conditions of 1-7% of salinity and pH 4-10, and treating with hot water bath to obtain broccoli-like Sb2S3；
The culture medium contains pentavalent antimony and S2O3 2-Inorganic salt medium of (4), or containing pentavalent antimony and SO3 2-The inorganic salt culture medium.
5. The process according to claim 4, wherein:
1) culturing the microorganism in a culture medium at a temperature of 10-50 deg.CoC. Reacting the mixture for 1-10 days in a dark place under the conditions of 1-7% of salinity and pH 4-10;
the culture medium contains 0.01-2mM of pentavalent antimony and 0.5-2mM of S2O3 2-Or inorganic salt medium containing 0.01-2mM of pentavalent antimony and 0.5-2mM of SO3 2-The inorganic salt medium of (1);
2) centrifuging the mixture obtained in step (1), placing the obtained precipitate in deionized water for ultrasonic treatment, resuspending the precipitate with 0.2-5% of anionic surfactant sodium dodecyl sulfate, and 70-100%oTreating with hot water bath for 1-45min, centrifuging, and freeze drying the precipitate to obtain powdery particles, i.e. biologically prepared cauliflower-like Sb2S3。
6. The process according to claim 4 or 5, wherein: the temperature is 20-40 deg.CoC. 1-3% salinity and pH 6-8.
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