CN114231520A - Carbon-based biological agent for efficiently converting heavy metal chromium in soil - Google Patents
Carbon-based biological agent for efficiently converting heavy metal chromium in soil Download PDFInfo
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- 239000011651 chromium Substances 0.000 title claims abstract description 115
- 239000002689 soil Substances 0.000 title claims abstract description 101
- VYZAMTAEIAYCRO-UHFFFAOYSA-N Chromium Chemical compound [Cr] VYZAMTAEIAYCRO-UHFFFAOYSA-N 0.000 title claims abstract description 74
- 229910052804 chromium Inorganic materials 0.000 title claims abstract description 73
- 229910052799 carbon Inorganic materials 0.000 title claims abstract description 47
- 239000003124 biologic agent Substances 0.000 title claims abstract description 46
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 title claims abstract description 44
- 229910001385 heavy metal Inorganic materials 0.000 title claims abstract description 25
- 241000894006 Bacteria Species 0.000 claims abstract description 22
- 244000005700 microbiome Species 0.000 claims abstract description 14
- 238000005067 remediation Methods 0.000 claims abstract description 10
- 238000002360 preparation method Methods 0.000 claims abstract description 9
- 239000003610 charcoal Substances 0.000 claims abstract description 8
- 238000012216 screening Methods 0.000 claims abstract description 7
- 239000002028 Biomass Substances 0.000 claims description 12
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 12
- 239000007787 solid Substances 0.000 claims description 12
- 239000002068 microbial inoculum Substances 0.000 claims description 11
- 241000196252 Ulva Species 0.000 claims description 9
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- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 claims description 4
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- 239000003183 carcinogenic agent Substances 0.000 description 1
- 238000009388 chemical precipitation Methods 0.000 description 1
- JOPOVCBBYLSVDA-UHFFFAOYSA-N chromium(6+) Chemical compound [Cr+6] JOPOVCBBYLSVDA-UHFFFAOYSA-N 0.000 description 1
- 239000000356 contaminant Substances 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
-
- 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
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F1/28—Treatment of water, waste water, or sewage by sorption
- C02F1/283—Treatment of water, waste water, or sewage by sorption using coal, charred products, or inorganic mixtures containing them
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- 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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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/02—Separating microorganisms from their culture media
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F2003/001—Biological treatment of water, waste water, or sewage using granular carriers or supports for the microorganisms
- C02F2003/003—Biological treatment of water, waste water, or sewage using granular carriers or supports for the microorganisms using activated carbon or the like
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/20—Heavy metals or heavy metal compounds
- C02F2101/22—Chromium or chromium compounds, e.g. chromates
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- Tropical Medicine & Parasitology (AREA)
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Abstract
The invention discloses a carbon-based biological agent for efficiently converting soil heavy metal chromium, and a preparation method and application thereof, and belongs to the technical field of soil remediation. The carbon-based biological agent comprises, by weight, 10-40 parts of charcoal, 60-90 parts of chromium reducing bacteria and a culture thereof; the carbon-based biological agent is prepared by the steps of preparing biochar, screening chromium reducing bacteria, mixing and fixing and the like, and is used for in-situ remediation of chromium-polluted soil. According to the invention, the biochar is taken as a carrier, the chromium reducing bacteria are fixed on the biochar, heavy metal chromium can be effectively adsorbed and converted, and Cr (VI) is reduced into Cr (III), so that the toxicity of the heavy metal is reduced. Meanwhile, the biochar is used as an excellent carrier of the microorganisms, so that the biochar can be uniformly distributed in the soil, and the viability of the microorganisms after entering the soil is improved, so that the synergistic effect of the biochar and the microbial carrier keeps the biological activity of the chromium reducing bacteria in the chromium-polluted soil, and the conversion efficiency of the chromium reducing bacteria on Cr (VI) is improved.
Description
Technical Field
The invention belongs to the technical field of soil remediation, and particularly relates to a carbon-based biological agent for converting soil heavy metal chromium, and a preparation method and application thereof.
Background
Chromium is a commercially important element used in steel making, leather tanning, chrome plating and paint manufacturing. Chromite is distributed throughout the world, with asia and africa being particularly widespread. It is estimated that mining 1 ton of chromite produces about 10 tons of chromium slag; the worldwide chromium slag stored annually is about 760 million tons, and 11.73 tons of Cr (VI) can be released into the environment. Chromium is commonly present in the environment in the form of hexavalent chromium (cr (vi)) and trivalent chromium (cr (iii)), which is recognized by the World Health Organization (WHO), the international agency for research on cancer, and the United States Environmental Protection Agency (USEPA) as a carcinogen harmful to humans because of the high toxicity of cr (vi). USEPA sets the limit of Cr (VI) discharge to surface water to 0.1 mg.L-1The drinking water limit is set to 0.05 mg.L-1. Soil is considered to be a collection of lipophilic organic contaminants and metals. CrO4 2-And Cr2O7 2-Is the main existing form of Cr (VI) in soil. Cr (vi) has strong mobility between soil and groundwater, thus presenting a significant challenge to the remediation of chromium-contaminated soil and water.
At present, the conventional methods for repairing chromium pollution comprise various methods, such as chemical precipitation, electrochemical separation, membrane filtration, ion exchange and the like, but the methods have the defects of high operation cost, ineffectiveness when the concentration of pollutant ions is low, secondary pollution and the like. In recent years, bioremediation, which is an eco-friendly chromium pollution remediation technology, has received much attention. Biological materials are used in bioremediation processes to reduce, eliminate, control and/or convert pollutants in water, soil, sediment and air. Among these biomaterials, microorganisms are widely used as suitable materials for bioremediation because they are ubiquitous and have a stronger growth ability and a faster contamination remediation efficiency. However, in the practical application process, the soil environment is complex and variable, the survival, growth and reproduction of microorganisms are influenced by the environment, and the microorganisms are sensitive to the environmental change, so that the bioremediation effect is directly influenced. How to maintain the biological activity of microorganisms in chromium-contaminated soil and how to improve the conversion efficiency of the microorganisms on Cr (VI) is a problem to be solved by applying the current biotechnology in chromium-contaminated soil remediation.
Disclosure of Invention
Aiming at the technical problems, the invention aims to provide a biological agent capable of efficiently converting soil heavy metal chromium, and a preparation method and application thereof. According to the invention, biochar is used as a carrier, and chromium reducing bacteria are fixed on the biochar to prepare the carbon-based chromium reducing biological agent. The biological agent can effectively adsorb and convert heavy metal chromium, and reduce Cr (VI) into Cr (III), thereby reducing the toxicity of heavy metal and realizing the remediation of polluted soil.
In order to achieve the purpose, the invention adopts the following technical scheme:
the invention provides a preparation method of a carbon-based biological microbial inoculum for efficiently converting heavy metal chromium in soil, which comprises the following steps:
(1) air-drying algae biomass, crushing, grinding, sieving by a sieve of 1-5mm to obtain biomass powder, putting the biomass powder into a crucible, compacting, covering, placing in a muffle furnace, raising the temperature to 600 ℃ at the speed of 3-5 ℃/min, keeping the temperature for 1-3h, naturally cooling, taking out, crushing, and sieving by a sieve of 50-100 meshes;
(2) sterilizing biochar at 121 ℃ for 15-30min, adding 10-40 parts by weight of biochar into an LB liquid culture medium, then inoculating 60-90 parts by weight of chromium reducing bacteria and culture thereof after overnight culture, and carrying out shake culture at constant temperature of 25-37 ℃ and at 130-170 rpm; after 24h, centrifuging at 3500-5500rpm for 5-15 min; the lower solid was washed with Tris-HCl at pH 9, centrifuged for 5-15min to remove the supernatant, and washed repeatedly 3-5 times with sterile nonwoven fabric to remove non-immobilized microorganisms.
Preferably, the algae biomass is one of enteromorpha and ulva.
Preferably, the optimal preparation temperature of the biochar is 400 ℃.
Preferably, the optimal part ratio of the biochar to the chromium reducing bacteria agent is 1: 4.
Further, the method for separating and screening the chromium reducing bacteria comprises the following steps:
and separating the chromium reducing bacteria by a dilution flat plate coating method. The Cr contaminated soil sample was transferred to a conical flask containing sterile water and shaken to homogenize the sample. Homogenized soil samples were serially diluted and spread on sterilized Luria Bertani (LB) agar plates. And (3) placing the LB solid culture medium after coating in a biochemical incubator for culture. Selecting Cr (VI) tolerant isolates with different colors and forms to be respectively cultured in LB liquid culture medium, measuring the Cr (VI) and total chromium content in the culture medium after 3-5 days, and screening out the strains with the best Cr (VI) reducing capability.
Further, the sample is shaken at 130-170rpm for 20-40min at room temperature.
Further, the composition of the LB solid medium comprises: 5-15g/L of peptone, 2-8g/L, NaCl 5-15g/L of yeast extract, 10-30g/L, pH ═ 7-10 of agar and 50-200mg/L of Cr (VI).
Further, the sterilization condition of the LB agar plate is high-pressure steam sterilization at 121 ℃ for 15-30 min.
Further, the LB solid medium is cultured in a biochemical incubator under the constant temperature of 25-35 ℃ for 3-5 d.
Further, the composition of the LB liquid medium comprises: 5-15g/L of peptone, 2-8g/L, NaCl 5-15g/L, pH-7-10 of yeast extract and 50-200mg/L of Cr (VI).
In a second aspect of the invention, the application of the carbon-based biological agent in chromium-contaminated soil remediation is provided. The application method comprises the following steps:
applying the prepared carbon-based biological agent to chromium-polluted soil, and fully ploughing to uniformly mix the agent and the soil; wherein the addition amount of the carbon-based biological agent is 0.5-3% of the weight of the soil.
For polluted soil with the total chromium content of 0-100mg/kg, the carbon-based biological agent is weighed according to 0.5% -1% of the soil weight and added into the soil, and 1% is preferred.
For the polluted soil with the total chromium content of 100-200mg/kg, the carbon-based biological agent is weighed according to 1-2% of the weight of the soil and is added into the soil, and the preferable weight is 2%.
For polluted soil with the total chromium content of more than 200mg/kg, the carbon-based biological agent is weighed according to 2% -3% of the soil weight and added into the soil, and 3% is preferred.
Compared with the prior art, the invention has the advantages that:
(1) the biochar serving as a carbon-rich material not only can be used as a soil conditioner to improve the physicochemical property of soil, but also can be used as a heavy metal adsorbent to repair heavy metal pollution in soil and water. The biochar in the carbon-based biological agent has certain adsorption and conversion capacity on Cr (VI).
(2) The biochar has the characteristics of high internal porosity, large specific surface area, rich functional groups, strong bacteria adsorption capacity and the like, and can be used as an excellent carrier of microorganisms to ensure that the microorganisms are uniformly distributed in the soil and improve the survival capacity of the microorganisms after entering the soil. The biochar in the carbon-based biological agent provided by the invention is cooperated with the chromium reducing bacteria, so that the biological activity in chromium-polluted soil is maintained, and the conversion efficiency of Cr (VI) is effectively improved.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
(1) Separating and screening of chromium reducing bacteria
And separating the chromium reducing bacteria by a dilution flat plate coating method. 10g of the Cr-contaminated soil sample was accurately weighed, transferred to a conical flask containing 100mL of sterile water, and homogenized by shaking at 140rpm for 30min at room temperature.
Preparing an LB solid culture medium, which comprises the following components: 10g/L of peptone, 5g/L, NaCl 10g/L of yeast extract, 20g/L, pH of agar-agar 9 and 100mg/L of Cr (VI). The medium was autoclaved at 121 ℃ for 20min, and then the homogenized soil sample was serially diluted and spread on LB agar plates. And placing the LB solid culture medium after finishing coating in a constant-temperature biochemical incubator at 30 ℃ for culture for 4 d.
After 4d, 4 Cr (VI) tolerant isolates with different staining and morphological parameters were selected and cultured in 50mL LB liquid medium separately. The LB liquid culture medium comprises: 10g/L of peptone, 5g/L, NaCl 10g/L, pH-9 of yeast extract and 100mg/L of Cr (VI). And 4d, measuring the Cr (VI) content and the total chromium content in the culture medium, thereby screening out the strain with the optimal Cr (VI) reduction capability.
TABLE 1 chromium removal rates of four different strains
As can be seen from Table 1, the strain LB-1 has the best Cr (VI) reducing ability. The strain LB-1 was identified as Bacillus cereus (Bacillus cereus) by 16S rDNA sequence analysis.
(2) Preparation of carbon-based biological agent
Preparing biochar:
and (3) drying the enteromorpha biomass, crushing, grinding and sieving by using a 2mm sieve to obtain enteromorpha powder. Placing the enteromorpha powder into a crucible, compacting and covering, placing the crucible in a muffle furnace, raising the temperature to 400 ℃ at the speed of 4 ℃/min, keeping the temperature for 2h, naturally cooling and taking out, crushing and sieving by a 60-mesh sieve to obtain the enteromorpha biochar.
Preparing a carbon-based biological agent:
weighing 20 parts by weight (1.5g) of enteromorpha biochar, sterilizing at 121 ℃ for 20min, cooling and then putting into 50mL of LB liquid culture medium. 80 parts by weight (6.0g) of chromium-reducing microbial inoculum after overnight culture was inoculated and cultured with shaking at a constant temperature of 30 ℃ and 140 rpm. Centrifuging at 5000rpm for 5min after 24 hr; the lower solid was washed with Tris-HCl (pH 9), centrifuged for 5min to remove the supernatant, and washed repeatedly 3 times with sterile nonwoven fabric to remove the non-immobilized microorganisms.
(3) Application and efficacy evaluation of carbon-based biological agent
The heavy metal contaminated soil is taken from a contaminated site of an electrochemical plant in Hangzhou city for carrying out an indoor pot experiment. The total chromium content in the soil of the site is 72.32mg/kg, the content of Cr (VI) is 25.61mg/kg, and the content of Cr (III) is 46.71 mg/kg. Taking 1kg of chromium-contaminated soil per pot, adding deionized water to keep the soil moisture at about 60% of the saturated water holding capacity, and balancing for 2 weeks for later use.
Four experiments of a blank control group (CK), a biochar group (BC), a chromium reducing microbial inoculum group (LB-1) and a carbon-based microbial inoculum group (BC + LB-1) are arranged together, wherein the CK group is not added with any repairing agent, and the rest groups are respectively added with biochar, chromium reducing microbial inoculum and carbon-based microbial inoculum which are 1 percent of the weight of the soil. And placing each group of soil for 45 days under the conditions of room temperature and natural illumination, and sampling after 45 days to determine the change of the chromium content of the soil.
After comparative analysis, the Cr (VI) content in the contaminated soil of each experimental group is reduced after the repairing agent is added, wherein the Cr (VI) content in the soil of the charcoal-based biological agent group (BC + LB-1) is reduced to the maximum extent of 82.27%, so that the repairing and converting effect of the charcoal-based biological agent prepared by compounding the biochar and the chromium reducing bacteria on the heavy metal chromium contaminated soil is obvious. The results of the specific examples are shown in Table 2.
Table 2 example 1 variation of chromium content in soil for different treatment groups
Example 2
The difference from example 1 is: the contents of total chromium, Cr (VI) and Cr (III) in the polluted soil are 175.30mg/kg, 68.42mg/kg and 106.88mg/kg respectively; biochar, a chromium reducing microbial inoculum and a carbon-based biological microbial inoculum which are 2 percent of the weight of the soil are respectively added into the soil in each experimental group.
After comparative analysis, the Cr (VI) content in the contaminated soil of each experimental group is reduced after the repairing agent is added, wherein the Cr (VI) content in the soil of the charcoal-based biological agent group (BC + LB-1) is reduced to the maximum extent of 73.91%, so that the repairing and converting effect of the charcoal-based biological agent prepared by compounding the biochar and the chromium reducing bacteria on the heavy metal chromium contaminated soil is obvious. The results of the specific examples are shown in Table 3.
Table 3 example 2 variation of chromium content in soil for different treatment groups
Example 3
The differences from the embodiments 1 and 2 are as follows: the contents of total chromium, Cr (VI) and Cr (III) in the polluted soil are 320.84mg/kg, 115.37mg/kg and 205.47mg/kg respectively; biochar, a chromium reducing microbial inoculum and a carbon-based biological microbial inoculum which are 3 percent of the weight of the soil are respectively added into the soil in each experimental group.
After comparative analysis, the Cr (VI) content in the contaminated soil of each experimental group is reduced after the repairing agent is added, wherein the Cr (VI) content in the soil of the charcoal-based biological agent group (BC + LB-1) is reduced to the maximum extent of 62.08%, so that the repairing and converting effect of the charcoal-based biological agent prepared by compounding the biochar and the chromium reducing bacteria on the heavy metal chromium contaminated soil is obvious. The results of the specific examples are shown in Table 4.
Table 4 example 3 variation of chromium content in soil for different treatment groups
The above description is only a preferred embodiment of the present application and is not intended to limit the present application, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application shall be included in the protection scope of the present application.
Claims (10)
1. A carbon-based biological agent for efficiently converting soil heavy metal chromium is characterized by consisting of biochar, chromium reducing bacteria and a culture thereof; the preparation method comprises the following steps:
(1) air-drying algae biomass, crushing, grinding, sieving by a sieve of 1-5mm to obtain biomass powder, putting the biomass powder into a crucible, compacting, covering, placing in a muffle furnace, raising the temperature to 600 ℃ at the speed of 3-5 ℃/min, keeping the temperature for 1-3h, naturally cooling, taking out, crushing, and sieving by a sieve of 50-100 meshes;
(2) sterilizing biochar at 121 ℃ for 15-30min, adding 10-40 parts by weight of biochar into an LB liquid culture medium, then inoculating 60-90 parts by weight of chromium reducing bacteria and culture thereof after overnight culture, and carrying out shake culture at constant temperature of 25-37 ℃ and at 130-170 rpm; after 24h, centrifuging at 3500-5500rpm for 5-15 min; the lower solid was washed with Tris-HCl at pH 9, centrifuged for 5-15min to remove the supernatant, and washed repeatedly 3-5 times with sterile nonwoven fabric to remove non-immobilized microorganisms.
2. A preparation method of a carbon-based biological agent for efficiently converting soil heavy metal chromium is characterized by comprising the following steps:
(1) air-drying algae biomass, crushing, grinding, sieving by a sieve of 1-5mm to obtain biomass powder, putting the biomass powder into a crucible, compacting, covering, placing in a muffle furnace, raising the temperature to 600 ℃ at the speed of 3-5 ℃/min, keeping the temperature for 1-3h, naturally cooling, taking out, crushing, and sieving by a sieve of 50-100 meshes;
(2) sterilizing biochar at 121 ℃ for 15-30min, adding 10-40 parts by weight of biochar into an LB liquid culture medium, then inoculating 60-90 parts by weight of chromium reducing bacteria and culture thereof after overnight culture, and carrying out shake culture at constant temperature of 25-37 ℃ and at 130-170 rpm; after 24h, centrifuging at 3500-5500rpm for 5-15 min; the lower solid was washed with Tris-HCl at pH 9, centrifuged for 5-15min to remove the supernatant, and washed repeatedly 3-5 times with sterile nonwoven fabric to remove non-immobilized microorganisms.
3. The carbon-based biological agent for efficiently converting soil heavy metal chromium according to claim 1, wherein the algae biomass is one of enteromorpha or ulva.
4. The carbon-based biological agent for efficiently converting soil heavy metal chromium according to claim 1, wherein the preparation temperature of the biochar is 400 ℃.
5. The carbon-based biological agent for efficiently converting soil heavy metal chromium according to claim 1, wherein the optimal part ratio of the biochar to the chromium reducing agent is 1: 4.
6. The carbon-based biological agent for efficiently converting soil heavy metal chromium according to claim 1, wherein the method for separating and screening chromium-reducing bacteria in the step (2) is as follows:
separating chromium reducing bacteria by a dilution flat plate coating method; transferring the Cr contaminated soil sample into a conical flask filled with sterile water, and oscillating to homogenize the sample; continuously diluting and coating the homogenized soil sample on a sterilized LB agar plate; placing the LB solid culture medium after coating in a biochemical incubator for culture; selecting Cr-tolerant isolates with different colors and forms to be respectively cultured in an LB liquid culture medium, measuring the Cr content and the total chromium content in the culture medium after 3-5 days, and screening out the strain with the optimal Cr reduction capability.
7. The carbon-based biological agent for efficiently converting soil heavy metal chromium according to claim 1, wherein the sample in the step (2) is oscillated at 130-170rpm for 20-40min at room temperature.
8. The charcoal-based biological microbial inoculum for efficiently transforming soil heavy metal chromium according to claim 1, wherein the LB solid culture medium comprises the following components: 5-15g/L of peptone, 2-8g/L, NaCl 5-15g/L of yeast extract, 10-30g/L, pH ═ 7-10 of agar and 50-200mg/L of Cr (VI);
the sterilization condition of the LB agar plate is high-pressure steam sterilization at 121 ℃ for 15-30 min;
the LB solid culture medium is cultured in a biochemical incubator under the constant temperature of 25-35 ℃ for 3-5 d;
the LB liquid culture medium comprises the following components: 5-15g/L of peptone, 2-8g/L, NaCl 5-15g/L, pH-7-10 of yeast extract and 50-200mg/L of Cr (VI).
9. The application of a carbon-based biological agent for efficiently converting soil heavy metal chromium in chromium-polluted soil remediation; the application method comprises the following steps:
applying the prepared carbon-based biological agent to chromium-polluted soil, and fully ploughing to uniformly mix the agent and the soil; wherein the addition amount of the carbon-based biological agent is 0.5-3% of the weight of the soil;
for polluted soil with the total chromium content of 0-100mg/kg, the carbon-based biological agent is weighed according to 0.5-1% of the weight of the soil and added into the soil;
for polluted soil with total chromium content of 100-200mg/kg, the carbon-based biological agent is weighed according to 1-2% of the weight of the soil and added into the soil;
and for the polluted soil with the total chromium content of more than 200mg/kg, weighing the carbon-based biological agent according to 2-3% of the weight of the soil, and adding the carbon-based biological agent into the soil.
10. The application of claim 8, which is as follows:
for polluted soil with the total chromium content of 0-100mg/kg, weighing the carbon-based biological agent according to 1% of the weight of the soil, and adding the carbon-based biological agent into the soil;
for polluted soil with the total chromium content of 100-200mg/kg, weighing the carbon-based biological agent according to 2% of the weight of the soil, and adding the carbon-based biological agent into the soil;
and for the polluted soil with the total chromium content of more than 200mg/kg, weighing the carbon-based biological agent according to 3% of the weight of the soil, and adding the carbon-based biological agent into the soil.
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