CN114196415A - Heavy metal passivator and application thereof - Google Patents
Heavy metal passivator and application thereof Download PDFInfo
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- CN114196415A CN114196415A CN202111643165.7A CN202111643165A CN114196415A CN 114196415 A CN114196415 A CN 114196415A CN 202111643165 A CN202111643165 A CN 202111643165A CN 114196415 A CN114196415 A CN 114196415A
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- 229910001385 heavy metal Inorganic materials 0.000 title claims abstract description 90
- 239000002689 soil Substances 0.000 claims abstract description 67
- 239000002068 microbial inoculum Substances 0.000 claims abstract description 64
- 239000007788 liquid Substances 0.000 claims abstract description 59
- 241000894006 Bacteria Species 0.000 claims abstract description 39
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims abstract description 39
- 239000004202 carbamide Substances 0.000 claims abstract description 39
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 claims abstract description 35
- 102000003846 Carbonic anhydrases Human genes 0.000 claims abstract description 29
- 108090000209 Carbonic anhydrases Proteins 0.000 claims abstract description 29
- 238000005067 remediation Methods 0.000 claims abstract description 25
- 230000033558 biomineral tissue development Effects 0.000 claims abstract description 23
- 238000002156 mixing Methods 0.000 claims abstract description 17
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims abstract description 13
- 239000000243 solution Substances 0.000 claims description 58
- 238000000855 fermentation Methods 0.000 claims description 30
- 230000004151 fermentation Effects 0.000 claims description 30
- 238000000034 method Methods 0.000 claims description 25
- 229940041514 candida albicans extract Drugs 0.000 claims description 13
- 239000012138 yeast extract Substances 0.000 claims description 13
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 12
- 230000001580 bacterial effect Effects 0.000 claims description 12
- 238000012258 culturing Methods 0.000 claims description 12
- 238000007865 diluting Methods 0.000 claims description 12
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 claims description 8
- 239000001888 Peptone Substances 0.000 claims description 7
- 108010080698 Peptones Proteins 0.000 claims description 7
- 239000007983 Tris buffer Substances 0.000 claims description 7
- 239000001963 growth medium Substances 0.000 claims description 7
- 235000019319 peptone Nutrition 0.000 claims description 7
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 6
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 6
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 6
- 239000011780 sodium chloride Substances 0.000 claims description 6
- 150000001875 compounds Chemical class 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 abstract description 4
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 10
- 241000193395 Sporosarcina pasteurii Species 0.000 description 10
- 230000000052 comparative effect Effects 0.000 description 7
- 230000008569 process Effects 0.000 description 6
- 229910002092 carbon dioxide Inorganic materials 0.000 description 5
- 239000001569 carbon dioxide Substances 0.000 description 5
- 150000001768 cations Chemical class 0.000 description 4
- 238000011081 inoculation Methods 0.000 description 4
- 229910052751 metal Inorganic materials 0.000 description 4
- 239000002184 metal Substances 0.000 description 4
- 238000007596 consolidation process Methods 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 230000008439 repair process Effects 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 101000965313 Legionella pneumophila subsp. pneumophila (strain Philadelphia 1 / ATCC 33152 / DSM 7513) Aconitate hydratase A Proteins 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 229910052500 inorganic mineral Inorganic materials 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 238000002386 leaching Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000011707 mineral Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000002161 passivation Methods 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 108010046334 Urease Proteins 0.000 description 1
- 238000005842 biochemical reaction Methods 0.000 description 1
- 238000010170 biological method Methods 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 238000009713 electroplating Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000000834 fixative Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 238000001095 inductively coupled plasma mass spectrometry Methods 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000002957 persistent organic pollutant Substances 0.000 description 1
- 238000000053 physical method Methods 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 238000007711 solidification Methods 0.000 description 1
- 230000008023 solidification Effects 0.000 description 1
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- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K17/00—Soil-conditioning materials or soil-stabilising materials
- C09K17/40—Soil-conditioning materials or soil-stabilising materials containing mixtures of inorganic and organic compounds
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C1/00—Reclamation of contaminated soil
- B09C1/08—Reclamation of contaminated soil chemically
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C1/00—Reclamation of contaminated soil
- B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2101/00—Agricultural use
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
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Abstract
The invention provides a heavy metal passivator and application thereof. The heavy metal passivator comprises a microbial inoculum, a stationary liquid and a urea solution; the microbial inoculum comprises a carbonate mineralization microbial inoculum and a carbonic anhydrase production microbial inoculum; the volume ratio of the carbonate mineralization bacterium liquid to the carbonic anhydrase producing bacterium liquid is (1-5) to 1; the stationary liquid is calcium chloride solution. The heavy metal passivator is applied to the remediation of heavy metal contaminated soil, and the remediation steps comprise: and uniformly mixing the microbial inoculum and the polluted soil, sequentially adding the stationary liquid and the urea solution, and fermenting for 7-8 days. The invention can ensure that the removal rate of the heavy metal reaches more than 90 percent only in 7-8 days, and greatly reduces the time for restoring the heavy metal contaminated soil.
Description
Technical Field
The invention relates to the technical field of heavy metal consolidation, in particular to a heavy metal passivator and application thereof.
Background
With the rapid development of the world economy, the pollution discharge problem in the process of the industrialization process is more and more serious, so that the heavy metal pollution problem is increasingly concerned globally. Unlike other organic pollutants, heavy metals in the environment are not easily degraded, and the heavy metals in soil and underground water around electroplating plants and battery plants can cause damage to the health of people and animals by permeating into a food chain, so that the existence of living things on the earth is threatened, and the balance of the ecological environment is damaged.
The heavy metal treatment method mainly comprises a chemical remediation method, a physical remediation method and a biological remediation method. The traditional physical and chemical remediation method has special requirements on equipment or materials in the process of treating heavy metal pollution, so that the remediation cost is increased, the remediation process is complex, and a large amount of manpower and material resources are required to be input. In recent years, biological treatment methods have attracted much attention because of their advantages such as high removal rate, low cost, no secondary pollution, and little environmental impact. The heavy metal mineral formed by the mineralization of the microorganisms has the advantages of large particle size, small mobility and stable structure, and has high application potential in the aspect of heavy metal pollution remediation of soil and underground water.
At present, the technology of inducing Carbonate Precipitation (MICP) by microorganisms to mineralize and consolidate heavy metals is mainly applied, and the MICP technology is based on a series of biochemical reactions of Carbonate mineralized bacteria, namely metal cations adsorbed to extracellular polymers of microorganisms and CO generated in the metabolic process of microorganisms3 2-Forming heavy metal minerals. However, at present, the carbonate mineralization bacteria are used for solidifying the heavy metals for a long time, and the time is basically more than 20 days for removing the heavy metalsThe rate reaches more than 90 percent, and the application of the carbonate mineralized bacteria in the consolidation of heavy metals is limited.
Disclosure of Invention
Therefore, the technical problem to be solved by the invention is to overcome the defect that the application of carbonate mineralization bacteria in the solidification of heavy metals is limited because the time consumption for solidifying the heavy metals by using the carbonate mineralization bacteria is long in the prior art.
Therefore, the invention provides a heavy metal passivator, which comprises a microbial inoculum, a stationary liquid and a urea solution; the microbial inoculum comprises a carbonate mineralization microbial inoculum and a carbonic anhydrase producing microbial inoculum; the volume ratio of the carbonate mineralization bacterium liquid to the carbonic anhydrase producing bacterium liquid is (1-5) to 1; the stationary liquid is calcium chloride solution.
Further, the method for obtaining the carbonate mineralization bacterium liquid comprises the following steps:
inoculating carbonate mineralized bacteria to NH4Culturing in-YE culture medium at 28-30 deg.C for 24-48 hr, and diluting to OD6001-1.2; and/or
The method for obtaining the carbonic anhydrase producing bacterial liquid comprises the following steps:
inoculating the strain of carbonic anhydrase producing bacteria into LB culture solution, culturing at 28-30 deg.C for 24-48h, and diluting to OD600The content of the compound is 1-1.2.
Further, said NH4The YE culture solution comprises 18-20g/L of yeast extract, 10-15g/L of ammonium sulfate and 13-16g/L of Tris (Tris (hydroxymethyl) aminomethane).
Further, the LB culture solution comprises 10-15g/L of peptone, 3-5g/L of yeast extract and 10-15g/L of sodium chloride.
Further, the concentration of the fixing solution is 0.01-0.05 mol/L.
Further, the concentration of the urea solution is 0.5-1.5 mol/L.
The invention provides an application of a heavy metal passivator, which is applied to remediation of heavy metal contaminated soil.
Further, the heavy metal contaminated soil remediation step comprises:
uniformly mixing the microbial inoculum with the contaminated soil in a volume ratio of (1-1.5) to 1, then sequentially adding a fixing solution with the contaminated soil in a volume ratio of (0.12-0.2) to 1 and a urea solution with the contaminated soil in a volume ratio of (1-5) to 1, and then fermenting for 7-8 days.
Further, standing for 2-5h after uniformly mixing the microbial inoculum and the polluted soil, and then adding the stationary liquid.
Further, adding an equal amount of urea solution every 48 hours after the fermentation is started; the fourth day after the start of fermentation, the same amount of the microbial inoculum and the same amount of the stationary liquid as before the start of fermentation were added once more.
The technical scheme of the invention has the following advantages:
1. according to the heavy metal passivator provided by the invention, the added carbonate mineralization bacteria secrete urease, decompose urea to form carbonate ions, and metabolize the carbonate ions to form carbonate precipitates to create a suitable alkaline environment, and meanwhile, the carbonate mineralization bacteria have negative charges and can adsorb metal cations with positive charges to cause metal cation enrichment, so that the carbonate ions and the metal cations form insoluble and stable carbonate precipitates by taking the carbonate mineralization bacteria as crystal nuclei, and thus heavy metals are passivated; the carbonic anhydrase producing bacteria can act together with the carbonate mineralization bacteria to accelerate the dissolution speed of carbon dioxide in a system, provide enough carbonate ions for the carbonate mineralization bacteria, greatly accelerate the speed of the carbonate mineralization bacteria for solidifying heavy metals, enable the removal rate of the heavy metals to reach more than 90% in 7-8 days, and greatly reduce the time for passivating the heavy metals.
2. According to the heavy metal passivator provided by the invention, the carbonic anhydrase producing bacteria can accelerate the heavy metal consolidation and consume part of carbon dioxide generated by the carbonate mineralization bacteria and the environment, so that the emission of carbon dioxide can be reduced, and in addition, the use of urea in the heavy metal passivation process is reduced.
3. The application method of the heavy metal passivator provided by the invention is mainly applied to the remediation of heavy metal contaminated soil, and compared with a chemical/physical method, the heavy metal passivator is non-toxic, harmless, free of secondary pollution, low in remediation cost, simple in remediation method, and capable of leaving a large amount of culture medium and urea in the soil in the remediation process, so that the soil is more fertile, and ecological remediation is more utilized.
4. The invention provides an application method of a heavy metal passivator, which firstly proposes to construct a mixed bacteria system of carbonate mineralized bacteria and carbonic anhydrase producing bacteria and is used for the biological method rapid repair of heavy metal contaminated soil.
Detailed Description
The following examples are provided to further understand the present invention, not to limit the scope of the present invention, but to provide the best mode, not to limit the content and the protection scope of the present invention, and any product similar or similar to the present invention, which is obtained by combining the present invention with other prior art features, falls within the protection scope of the present invention.
The examples do not show the specific experimental steps or conditions, and can be performed according to the conventional experimental steps described in the literature in the field. The reagents or instruments used are not indicated by manufacturers, and are all conventional reagent products which can be obtained commercially.
The carbonate mineralization bacteria is Bacillus pasteurii, collection number ATCC 11895, and is purchased from American type culture Collection.
The carbonic anhydrase producing bacteria are shown carbonic anhydrase on cell surface and its enzymological property research according to the literature [ D ]]Tianjin university. "constructed E-22b-IRLAnd (3) strain S.
The yeast extract was purchased from Oboxing Biotechnology, Inc., Beijing.
The peptone used was purchased from Oboxing Biotechnology, Inc., Beijing.
Tris used was purchased from McClin Biotechnology Ltd.
The heavy metal contaminated soil is from a heavy metal contaminated site in Shandong.
Example 1
The embodiment provides a heavy metal passivator, which comprises a microbial inoculum, a stationary liquid and a urea solution; the microbial inoculum comprises a carbonate mineralization microbial inoculum and a carbonic anhydrase production microbial inoculum; the stationary liquid is calcium chloride solution; the heavy metal passivator is applied to remediation of heavy metal contaminated soil; the specific application method comprises the following steps:
obtaining a microbial inoculum: inoculation of Bacillus pasteurii into NH4Culturing in-YE culture medium at 30 deg.C for 24 hr, and diluting to OD600Obtaining a bacillus pasteurii bacterial liquid as 1; inoculating carbonic anhydrase producing bacteria into LB culture solution, culturing at 30 deg.C for 24 hr, and diluting to OD600Obtaining carbonic anhydrase producing bacteria liquid as 1; uniformly mixing the bacillus pasteurii bacterial liquid and the carbonic anhydrase producing bacterial liquid according to the volume ratio of 1:1 to obtain a microbial inoculum; NH used4YE culture solution comprises yeast extract 20g/L, ammonium sulfate 10g/L and Tris 15.748 g/L; the LB culture solution comprises 10g/L of peptone, 5g/L of yeast extract and 10g/L of sodium chloride;
adding a microbial inoculum: taking heavy metal contaminated soil, adding a microbial inoculum into the heavy metal contaminated soil, uniformly mixing, and standing for 3h, wherein the volume ratio of the added microbial inoculum to the contaminated soil is 1: 1;
adding a stationary liquid: adding a calcium chloride solution with the concentration of 0.04mol/L into the standing soil and uniformly mixing; the volume ratio of the added fixing liquid to the soil is 0.15: 1;
adding a urea solution: adding a 1mol/L urea solution, wherein the volume ratio of the added urea solution to soil is 1: 1; then fermentation is started, and the heavy metal ions in the soil can be solidified after fermentation for 7 days;
adding an equal amount of urea solution every 48 hours after the fermentation is started; the fourth day after the start of fermentation, the same amount of the microbial inoculum and the same amount of the stationary liquid as before the start of fermentation were added once more.
Example 2
The embodiment provides a heavy metal passivator, which comprises a microbial inoculum, a stationary liquid and a urea solution; the microbial inoculum comprises a carbonate mineralization microbial inoculum and a carbonic anhydrase production microbial inoculum; the stationary liquid is calcium chloride solution; the heavy metal passivator is applied to remediation of heavy metal contaminated soil; the specific application method comprises the following steps:
obtaining a microbial inoculum: inoculation of Bacillus pasteurii into NH4Culturing in-YE culture medium at 30 deg.C for 24 hr, and diluting to OD6001.2, obtaining a bacillus pasteurii bacterial liquid; inoculating carbonic anhydrase producing bacteria into LB culture solution, culturing at 28 deg.C for 48h, and diluting to OD600Obtaining carbonic anhydrase producing bacteria liquid as 1; uniformly mixing the bacillus pasteurii bacterial liquid and the carbonic anhydrase producing bacterial liquid according to the volume ratio of 5:1 to obtain a microbial inoculum; NH used4YE culture solution comprises yeast extract 18g/L, ammonium sulfate 15g/L and Tris 13 g/L; the LB culture solution comprises peptone 15g/L, yeast extract 3g/L and sodium chloride 10 g/L;
adding a microbial inoculum: taking heavy metal contaminated soil, adding a microbial inoculum into the heavy metal contaminated soil, uniformly mixing, standing for 2h, wherein the volume ratio of the added microbial inoculum to the contaminated soil is 1: 1;
adding a stationary liquid: adding a calcium chloride solution with the concentration of 0.01mol/L into the settled soil and uniformly mixing; the volume ratio of the added fixing liquid to the soil is 0.2: 1;
adding a urea solution: adding a urea solution with the concentration of 0.5mol/L, wherein the volume ratio of the added urea solution to the soil is 1: 1; then starting fermentation for 8 days to passivate heavy metal ions in the soil;
adding an equal amount of urea solution every 48 hours after the fermentation is started; the fourth day after the start of fermentation, the same amount of the microbial inoculum and the same amount of the stationary liquid as before the start of fermentation were added once more.
Example 3
The embodiment provides a heavy metal passivator, which comprises a microbial inoculum, a stationary liquid and a urea solution; the microbial inoculum comprises a carbonate mineralization microbial inoculum and a carbonic anhydrase production microbial inoculum; the stationary liquid is calcium chloride solution; the heavy metal passivator is applied to remediation of heavy metal contaminated soil; the specific application method comprises the following steps:
obtaining a microbial inoculum: inoculation of Bacillus pasteurii into NH4Culturing in-YE culture medium at 28 deg.C for 48h, and diluting to OD600Obtaining a bacillus pasteurii bacterial liquid as 1; inoculating carbonic anhydrase producing bacteria into LB culture solution, culturing at 30 deg.C for 24 hr, and diluting to OD600Obtaining carbonic anhydrase producing bacteria liquid as 1.2; uniformly mixing the pasteur bacillus bacterial liquid and carbonic anhydrase producing bacterial liquid according to the volume ratio of 2:1 to obtain bacteriaAn agent; NH used4YE culture solution comprises yeast extract 20g/L, ammonium sulfate 10g/L and Tris 16 g/L; the LB culture solution comprises 10g/L of peptone, 5g/L of yeast extract and 15g/L of sodium chloride;
adding a microbial inoculum: taking heavy metal contaminated soil, adding a microbial inoculum into the heavy metal contaminated soil, uniformly mixing, standing for 5 hours, wherein the volume ratio of the added microbial inoculum to the contaminated soil is 1.5: 1;
adding a stationary liquid: adding a calcium chloride solution with the concentration of 0.05mol/L into the settled soil and uniformly mixing; the volume ratio of the added fixing liquid to the soil is 0.12: 1;
adding a urea solution: adding a 1.5mol/L urea solution, wherein the volume ratio of the added urea solution to soil is 1: 1; then, fermentation is started, and the heavy metal passivation in the soil can be completed after 7 days of fermentation;
adding an equal amount of urea solution every 48 hours after the fermentation is started; the fourth day after the start of fermentation, the same amount of the microbial inoculum and the same amount of the stationary liquid as before the start of fermentation were added once more.
Comparative example 1
The comparative example provides a heavy metal passivator, comprising a microbial inoculum, a stationary liquid and a urea solution; the microbial inoculum comprises carbonate mineralized bacteria liquid; the stationary liquid is calcium chloride solution; the heavy metal passivator is applied to remediation of heavy metal contaminated soil; the specific application method comprises the following steps:
obtaining a microbial inoculum: inoculation of Bacillus pasteurii into NH4Culturing in-YE culture medium at 30 deg.C for 24 hr, and diluting to OD600Obtaining the microbial inoculum after 1; NH used4YE culture solution comprises yeast extract 20g/L, ammonium sulfate 10g/L and Tris 15.748 g/L;
adding a microbial inoculum: taking heavy metal contaminated soil, adding a microbial inoculum into the heavy metal contaminated soil, uniformly mixing, and standing for 3h, wherein the volume ratio of the added microbial inoculum to the contaminated soil is 1: 1;
adding a stationary liquid: adding a calcium chloride solution with the concentration of 0.04mol/L into the standing soil and uniformly mixing; the volume ratio of the added fixing liquid to the soil is 0.15: 1;
adding a urea solution: adding a 1mol/L urea solution, wherein the volume ratio of the added urea solution to soil is 1: 1; then fermentation is started, and the remediation of the heavy metal contaminated soil can be completed after 7 days of fermentation;
adding an equal amount of urea solution every 48 hours after the fermentation is started; the fourth day after the start of fermentation, the same amount of the microbial inoculum and the same amount of the stationary liquid as before the start of fermentation were added once more.
Comparative example 2
The comparative example provides a heavy metal passivator, comprising a microbial inoculum, a stationary liquid and a urea solution; the microbial inoculum comprises carbonic anhydrase producing bacterial liquid; the stationary liquid is calcium chloride solution; the heavy metal passivator is applied to remediation of heavy metal contaminated soil; the specific application method comprises the following steps:
obtaining a microbial inoculum: inoculating carbonic anhydrase producing bacteria into LB culture solution, culturing at 30 deg.C for 24 hr, and diluting to OD600Obtaining the microbial inoculum after 1; the LB culture solution comprises 10g/L of peptone, 5g/L of yeast extract and 10g/L of sodium chloride;
adding a microbial inoculum: taking heavy metal contaminated soil, adding a microbial inoculum into the heavy metal contaminated soil, uniformly mixing, and standing for 3h, wherein the volume ratio of the added microbial inoculum to the contaminated soil is 1: 1;
adding a stationary liquid: adding a calcium chloride solution with the concentration of 0.04mol/L into the standing soil and uniformly mixing; the volume ratio of the added fixing liquid to the soil is 0.15: 1;
adding a urea solution: adding a 1mol/L urea solution, wherein the volume ratio of the added urea solution to soil is 1: 1; then fermentation is started, and the remediation of the heavy metal contaminated soil can be completed after 7 days of fermentation;
adding an equal amount of urea solution every 48 hours after the fermentation is started; the fourth day after the start of fermentation, the same amount of the microbial inoculum and the same amount of the stationary liquid as before the start of fermentation were added once more.
Comparative example 3
The comparative example differs from example 1 only in that the fixative was not added and the remaining conditions were maintained.
Test example 1
Taking 100g of soil before remediation, soaking the soil in 1000mL of deionized water, filtering to obtain a leaching solution, and detecting the content of heavy metals in the leaching solution by ICP-MS; according to the method, the heavy metal content in the soil repaired in the examples 1 to 3 and the comparative examples 1 to 3 is detected, and compared with the soil before repair, the less the heavy metal content is, the higher the heavy metal removal rate is, and the test result is shown in table 1:
TABLE 1 heavy metal content
As can be seen from Table 1, the two-bacterium system in the application can enable the heavy metal removal rate to reach more than 90% only in 7-8 days, greatly reduces the time for repairing the heavy metal contaminated soil compared with the traditional single-bacterium system repair, and simultaneously utilizes the carbon dioxide in the system, reduces the emission of the carbon dioxide and reduces the dosage of the urea.
It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications therefrom are within the scope of the invention.
Claims (10)
1. The heavy metal passivator is characterized by comprising a microbial inoculum, a stationary liquid and a urea solution; the microbial inoculum comprises a carbonate mineralization microbial inoculum and a carbonic anhydrase producing microbial inoculum; the volume ratio of the carbonate mineralization bacterium liquid to the carbonic anhydrase producing bacterium liquid is (1-5) to 1; the stationary liquid is calcium chloride solution.
2. The heavy metal passivator of claim 1, wherein the method for obtaining the carbonate mineralization bacteria liquid comprises:
inoculating carbonate mineralized bacteria to NH4Culturing in-YE culture medium at 28-30 deg.C for 24-48 hr, and diluting to OD6001-1.2; and/or
The method for obtaining the carbonic anhydrase producing bacterial liquid comprises the following steps:
inoculating the strain of carbonic anhydrase producing bacteria into LB culture solution, culturing at 28-30 deg.C for 24-48h, and diluting to OD600The content of the compound is 1-1.2.
3. The heavy metal passivator of claim 2, wherein the NH is4The YE culture solution comprises 18-20g/L of yeast extract, 10-15g/L of ammonium sulfate and 13-16g/L of Tris.
4. The heavy metal passivator of claim 2, wherein the LB culture solution comprises peptone 10-15g/L, yeast extract 3-5g/L and sodium chloride 10-15 g/L.
5. The heavy metal passivator of any one of claims 1-4, wherein the concentration of the fixing solution is 0.01-0.05 mol/L.
6. A heavy metal passivator according to any one of claims 1 to 5 wherein the concentration of the urea solution is 0.5 to 1.5 mol/L.
7. The application of the heavy metal passivator is characterized in that the heavy metal passivator is the heavy metal passivator as claimed in any one of claims 1 to 6, and is applied to remediation of heavy metal contaminated soil.
8. The use according to claim 7, wherein the step of remediating heavy metal contaminated soil comprises:
uniformly mixing the microbial inoculum with the contaminated soil in a volume ratio of (1-1.5) to 1, then sequentially adding a fixing solution with the contaminated soil in a volume ratio of (0.12-0.2) to 1 and a urea solution with the contaminated soil in a volume ratio of (1-5) to 1, and then fermenting for 7-8 days.
9. The application of claim 8, wherein the fixing solution is added after the microbial inoculum and the polluted soil are mixed uniformly and are kept stand for 2-5 h.
10. Use according to claim 8, characterized in that an equal amount of urea solution is added every 48h after the start of the fermentation; the fourth day after the start of fermentation, the same amount of the microbial inoculum and the same amount of the stationary liquid as before the start of fermentation were added once more.
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| CN115433687A (en) * | 2022-09-28 | 2022-12-06 | 华中科技大学 | Engineering bacterium for removing heavy metal ions through broad-spectrum and high-efficiency adsorption and coupling biomineralization |
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| CN103951532A (en) * | 2014-05-21 | 2014-07-30 | 安徽飞天农用生物科技股份有限公司 | Compound soil remediation agent containing microorganism bacteria and preparation method thereof |
| CN108220380A (en) * | 2018-02-08 | 2018-06-29 | 天津科技大学 | The method for preparing calcium carbonate using two kinds of common mineralisings of microorganism |
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| CN103951532A (en) * | 2014-05-21 | 2014-07-30 | 安徽飞天农用生物科技股份有限公司 | Compound soil remediation agent containing microorganism bacteria and preparation method thereof |
| CN108220380A (en) * | 2018-02-08 | 2018-06-29 | 天津科技大学 | The method for preparing calcium carbonate using two kinds of common mineralisings of microorganism |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN115433687A (en) * | 2022-09-28 | 2022-12-06 | 华中科技大学 | Engineering bacterium for removing heavy metal ions through broad-spectrum and high-efficiency adsorption and coupling biomineralization |
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