Method for preparing a bacterial colony complex purifying agent for synergistic treatment of PAEs and Cd.
A bacterial colony complex purifying agent using modified montmorillonite and biochar carriers with Cd-resistant strains addresses the inefficiencies in treating PAEs and Cd contamination by enhancing adsorption and passivation, achieving simultaneous removal of both contaminants from agricultural soils.
Patent Information
- Authority / Receiving Office
- JP · JP
- Patent Type
- Applications
- Current Assignee / Owner
- NANJING AGRICULTURAL UNIVERSITY
- Filing Date
- 2024-08-12
- Publication Date
- 2026-06-04
AI Technical Summary
Current methods for treating combined contamination of phthalate esters (PAEs) and heavy metals like cadmium (Cd) in agricultural soils, particularly in karst regions, are inefficient due to insufficient adsorption and passivation capacity, and the bonding coefficient is small, leading to desorption issues.
A method is developed to prepare a bacterial colony complex purifying agent using modified montmorillonite with mercapto groups and biochar carriers, enhancing the passivation of heavy metals and the degradation of PAEs by incorporating Cd-resistant bacterial strains that form interdependent relationships through swarm sensing mechanisms.
The method effectively removes both PAEs and Cd from soil by leveraging the synergistic action of modified montmorillonite and biochar carriers with Cd-resistant bacterial strains, improving adsorption and passivation, thus overcoming the limitations of existing treatments.
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Abstract
Description
[Technical Field]
[0001] The present invention relates to the technical field of soil contaminant treatment, and more specifically to the synergistic treatment of PAEs and Cd. This invention relates to a method for preparing a bacterial colony complex purifying agent for control purposes. [Background technology]
[0002] In agricultural soils in karst regions, phthalate ester (PAEs) plasticizers and heavy metals Cd The combined contamination is becoming a problem. To address the above problem, currently, a complex consisting of functional bacterial colonies and their carriers is being developed. Using immobilized bacterial agents is a cost-effective method. To address the problem of combined contamination of phthalate plasticizers and heavy metals Cd, bacterial colonization —It is necessary for the cells to withstand Cd's attack and for PAEs to be efficiently removed. The inventors efficiently degrade PAEs using a swarm sensing mechanism based on the decomposition capacity of the bacterial strain. We constructed functional bacterial colonies that can perform the necessary functions. When preparing the carrier for the functional bacterial colonies described above, the inventors found the prior art to have the following problems. I discovered that... In managing PAEs problems in agricultural land, bacterial colonies and biochar are currently being used. The carriers show good effects in both adsorption and removal of organic matter, but both are effective against heavy metals. The adsorption and passivation capacity is insufficient, the bonding coefficient is small, and it is prone to desorption. It is present. To solve the above problem, the present inventors have developed a method for modifying Montmorillonina with mercapto. By introducing threads, the passivation ability of the purification agent against heavy metals is improved, and furthermore, bacterial colonies Designed a fixed-type composite remediation agent and phthalate ester plasticizer for agricultural soils in karst regions. It also provides technical support to solve the problem of excessive combined contamination by heavy metals such as cadmium. [Overview of the project]
[0003] To achieve the above objective, the present invention provides a detailed method for synergistically processing PAEs and Cd. This invention provides a method for preparing a bacterial colony complex purifying agent, and the complex purifying agent prepared by this method is suitable for use in machinery. Potent bacterial colonies not only efficiently decompose PAEs in the soil, but also mercaptolytically break down PAEs. The carrier material passiveizes Cd in the soil, and simultaneously removes PAEs and Cd from the soil. It is possible to achieve this. A bacterial colony complex designed according to the present invention for synergistically treating PAEs and Cd. The preparation method for the purifying agent includes the following steps: S1, Prepare functional bacterial colonies. S1-1, Domestication of Cd-resistant strains: Normal growth of strains and the function of strains that degrade PAEs. Under the condition that we guarantee the following, strains of the genera Goldonella, Erythrobacter, and Bacillus are grown in a gradient concentration. Cd 2+ They were induced to become domesticated using LB medium. The three strains treated with S1-2 and S1-1 were activated in LB medium for 24 hours, and OD 600 =1. Adjust to 0, S1-3, the three strains treated with S1-2 were mixed in a volume ratio of 1:1:1, and the PAEs were divided. We prepared functional bacterial colonies that possess both resorption and Cd resistance functions. S2. Prepare the bacterial colony carrier. S2-1, mercaptomontmorillonite was prepared, S2-2, prepare a biochar carrier, S2-3, carrier composite: First, mercaptomontmorillonite in S2-1 and bi in S2-2 The charcoal carrier is placed in pure water, mixed uniformly, then centrifuged, washed, and dried to remove bacterial colonies. Gaining a body, S3. Prepare a bacterial colony composite purifying agent. S3-1. Uniformly mix the functional bacterial colony prepared in S1 and the bacterial colony carrier prepared in S2 to obtain a mixed system. Let n be the magnification factor, where n ∈ R + Then, the addition amount of the bacterial colony carrier is 1n g, and the addition amount of the functional bacterial colony is [10n, 30n] mL. S3-2. First, culture the mixed system obtained in S3-1 at 150 - 200 r / min, at a constant temperature of 30 °C, under light-shielding conditions for 24 h with shaking. Then, under sterile conditions, after filtration and rinsing, transfer it to drying at 25 °C to prepare a bacterial colony composite purifying agent. Furthermore, the mercapto montmorillonite in S2-1 is sodium silicate-mercapto montmorillonite or polyvinyl alcohol-mercapto montmorillonite. Explanation: Generally, the interlayer spacing of the montmorillonite structure is less than 1 nm as it is. It is difficult to adsorb and fix montmorillonite and the biochar carrier, and it is difficult to form a stable organic-mineral composite structure, and there is a risk of falling off during use. Therefore, it is necessary to expand the interlayer gap of montmorillonite. Furthermore, the preparation method of sodium silicate-mercapto montmorillonite is as follows : S2-1-A. Add montmorillonite and sodium silicate to an ethanol-aqueous solution of 3-mercaptopropyltrimethoxysilane, mix uniformly, and then control the temperature at 25 °C and the pH at 9 .5 - 10, react for 6 - 7 h while continuing stirring, and prepare small-pore montmorillonite modified with mercapto, denoted as sodium silicate-mercapto montmorillonite . Here, montmorillonite, sodium silicate mercapto compound, and 3-mercaptoprop The mass ratio of ropiltrimethoxysilane is 1:0.06:0.1. The mass concentration of the ethanol-aqueous solution is 95%. The mass sum of montmorillonite and sodium silicate is M1,3-mercaptopropyltrimeth If V1 is the sum of the volumes of xysilane and ethanol aqueous solution, then M1:V1 = 1.06g:2 It is 0 mL. Explanation: After sodium modification, the original Ca between the montmorillonite layers 2+ Na + Replaced with As a result, the montmorillonite particles become even smaller, and stoneflake-like characteristics become more pronounced. As a result, the adsorption capacity improved, and the ion exchange capacity within the interlayer domains of the crystal cell improved. Furthermore, the method for preparing polyvinyl alcohol-mercaptomontmorillonite is as follows: can be: S2-1-B-1, Interlayer insertion of organic matter: First, montmorillonite is dissolved in deionized water and the Prepare a suspension with a mass concentration of 2-4%, then add PVA to deionized water to a mass concentration of 2-3%. Prepare a % mixture, then add the solution to the suspension in a volume ratio of 1:4, and vibrate with an ultrasonic vibrator for 5-7 days. Then, obtain a colloidal suspension. S2-1-B-2, Low-temperature self-assembly: Processing of the colloidal suspension obtained in S2-1-B-1. After that, freeze at -10°C for 24 hours to expand the interlayer spacing and then extract the large interlayer montmorillonite. Gain, S2-1-B-3, Mercapto-modified: Large intercalated montmorillonite in S2-1-B-2 Add to an ethanol-aqueous solution of 3-mercaptopropyltrimethoxysilane and mix uniformly. After mixing, control the temperature to 25°C and the pH to 9.5-10, and continue stirring while reacting for 6-7 hours. Then, large intercalated montmorillonite, modified by mercapto, was prepared, and polyvinyl It is written as nyl alcohol-mercaptomontmorillonite, Here, large intercalated montmorillonite and 3-mercaptopropyltrimethoxysilane The mass ratio is 1:0.1, The mass concentration of the ethanol-aqueous solution is 95%. The mass of large intercalated montmorillonite is M2,3-mercaptopropyltrimethoxysilane If V2 is the sum of the volumes of the ethanol-aqueous solution, then M2:V2 = 1g:20mL. Description: The original montmorillonite has a small interlayer spacing, making it stable with the biochar material used in subsequent processes. Because this is unfavorable for bonding, the interlayer spacing of each layer of montmorillonite needs to be increased: When preparing polyvinyl alcohol-mercaptomontmorillonite, PVA is used. It is inserted into the montmorillonite gap, and the montmorillonite gap is expanded by low-temperature self-assembly. It provides the basis for subsequent bonding. The principle of low-temperature self-assembly of montmorillonite modified with PVA is as follows: At high temperatures, it freezes and forms ice crystals, and at the solid-liquid interface, the solute separates from the ice crystals, and the ice crystals further form montmorillonite. The lonite particles are concentrated in the gaps of the ice crystals, slowly assemble under the action of supramolecules, and finally freeze. After drying, a large interlaminar montmorillonite material with regularly distributed pores is obtained. Furthermore, the interlayer insertion method for S2-1-B-1, which involves organic matter, is as follows: S2-1-B-1-1, First, dissolve montmorillonite in deionized water to a mass concentration of 2-4%. Prepare a suspension and stir at 45-50°C for 3-4 hours, then add PVA to deionized water. Prepare a mixed solution with a mass concentration of 2-3%, Add solution S2-1-B-1-2 to the suspension in a volume ratio of 1:4, and follow the parameters below. Insert PVA into the montmorillonite gap: First, vibrate with ultrasound for 3-5 minutes and let stand for 1 hour, then vibrate with ultrasound for 4-5 minutes. Let stand for 10 hours, then shake with ultrasound for 5 minutes and let stand for 10 hours, followed by stirring at 45-50°C for 3 hours. Mix the mixture, then seal it and shake it ultrasonically at room temperature for 5-7 days to obtain a colloidal suspension. The colloidal suspensions in S2-1-B-1-3 and S2-1-B-1-2 were separated by centrifugation. The separated material is obtained, and the separated material is repeatedly washed with deionized water, and finally the separated material is diluted with deionized water. I'll keep it as a spare. Furthermore, biochars paired with sodium silicate-mercaptomontmorillonite The body is made of biocarbon particles, and is paired with polyvinyl alcohol-mercaptomontmorillonite. The biochar carrier used is a string-like carbon fiber. Explanation: Because the biochar particles have a small particle size and a large specific surface area, they bond with montmorillonite. This makes it easier, but at the same time, the bonding surface with montmorillonite is struck, so the bonding with bacterial colonies is also affected. The surface area becomes smaller, and as a result, the amount of bacterial colony that can be carried decreases, and the lattice-like carbon fiber becomes biochar Unlike particles, its specific surface area is smaller than that of biochar particles, but it has a larger absolute surface area. And while one end binds to montmorillonite, the rest still contains numerous bacterial colonies. There is room for support, but as a result, the bonding strength with montmorillonite decreases. Furthermore, the method for preparing biochar particles is as follows: After crushing and polishing plant fibers, inactivation The product is thermally decomposed under a thermal atmosphere, and after thermal decomposition, the product is polished and passed through a 200-mesh sieve to bio-grade materials. Prepare carbon particles, The parameters for pyrolysis are as follows: the furnace temperature is increased from room temperature to 60°C at a rate of 4-5°C / min. Raise the temperature to 0°C and maintain it for 2 hours. Description: Plant fibers include, without particular limitation, plant fibers such as corn and rice straw. You can choose from these options, and biochar particles can be used to improve the binding strength during loading. It just needs to be small and uniform. Furthermore, the method for preparing the stratified carbon fibers is as follows: First, 1-2 mm long lignocellulose is immersed in a 0.1 mol / L NaOH solution, 5 Stir at 00-550 r / min for 20-25 hours, then remove the lignocellulose. The material is then dried at 80°C, followed by firing at a gradient temperature. After firing, the product is washed and dried to form a rope-like structure. Obtain carbon fiber, The temperature gradient is as follows: First, it rises from 20°C to 250°C at a rate of 4-5°C / min. Heat, maintain temperature for 5-6 minutes, then heat at a rate of 20-22°C / min from 250°C to 500°C. Heat to ℃, maintain the temperature for 1-2 hours, and finally cool to 20℃ in a furnace. Description: Lignocellulose can retain its plant form even after high-temperature firing, and plant fiber bone It forms a lattice-like fiber structure. Furthermore, the method for composite formation in S2-3 is as follows: S2-3-1, Mercaptomontmorillonite obtained in S2-1 and biochar in S2-2 The carrier is placed in pure water and mixed uniformly to obtain a mixture, and n is the multiplication coefficient, where n∈R + So, The amount of mercaptomontmorillonite added is 1 ng, and the amount of biochar support added is [3 ng] The amount of pure water added is [100n, 110n] g, and the amount of pure water added is [4n] mL. The mixture obtained in S2-3-2 and S2-3-1 was separated, and the separated product was washed. The material is dried at 100°C for 5 hours, and finally passed through a 200-mesh sieve to obtain a composite support. Explanation: Taking lignocellulose as an example, after carbonization, the lignocellulose retains the original plant form. It has a string-like skeleton, and after mixing with polyvinyl alcohol-mercaptomontmorillonite, the string The carbon fibers are bonded to polyvinyl alcohol-mercaptomontmorillonite, and on their surface The intertwined, rope-like carbon fiber surface and its gaps can provide numerous bonding sites. It provides structural support for subsequent adhesion of bacterial strains. Compared to existing bacterial agents for degrading PAEs, the present invention has the following beneficial effects. (1) The composite purifying agent designed according to the present invention has functional bacterial colonies that are cadmium resistant. By being domesticated to possess this characteristic, they have broad degradability against PAEs. However, each degrading strain within a bacterial colony can form interdependent relationships through group sensing. A single functional bacterial strain exhibits reduced decomposition efficiency under heavy metal Cd contamination stress in complex contaminated soil. This allows us to overcome the problem of weakened competitiveness. (2) The composite purifying agent designed according to the present invention is modified by mercapto and cadmium It has an excellent passivation effect against heavy metals such as, and in cooperation with functional bacterial colonies, This method can achieve the effect of simultaneously removing PAEs and Cd from the soil. [Brief explanation of the drawing]
[0004] [Figure 1] This is a growth curve diagram of the cadmium-resistant domesticated bacterial strain in Example 1 from 0 to 48 hours, where Gordonia sp. is the genus Gordonia, Rhodococcus sp. is the genus Erythrobacter, and Bacillus sp. is the genus Bacillus. [Figure 2] This figure shows the OD600 values of cadmium-resistant domesticated bacterial strains grown for 24 hours in LB medium with different Cd2+ concentrations in Example 1. [Figure 3] This figure shows the OD600 values of cadmium-resistant domesticated bacterial strains grown for 24 hours in LB medium at different pH levels in Example 1. [Figure 4] This figure shows the degradation rates of the cadmium-resistant domesticated bacterial strains against six types of PAEs in Example 1. [Figure 5]This figure shows the saturated adsorption amount of Cd2+ by the bacterial colony carrier in Example 4, Comparative Example 2, and Comparative Example 3. Here, error bars a and b are letter markers in SPSS significance analysis, indicating significant differences in the PAEs degradation rate by the bacterial agent. [Figure 6] This figure shows the degradation efficiency of the combined purifying agent after 5d culture in Examples 4-10 and Comparative Examples 1-3. Here, a2 is Example 4, a3 is Example 5, a4 is Example 6, a5 is Example 7, a6 is Example 8, a7 is Example 9, a8 is Example 10, b1 is Comparative Example 1, b2 is Comparative Example 2, and b3 is Comparative Example 3. The error bars a, b, d, f, ef, e, cd, and bc are all letter markers in SPSS significance analysis, indicating significant differences in PAE degradation rates by bacterial agents. [Figure 7] This is an SEM morphological view of the composite purifying agent in Example 4. [Figure 8] This diagram shows the decomposition efficiency of PAEs in soil by the combined remediation agent in Example 4. [Figure 9] This figure shows the passivation efficiency of the heavy metal Cd by the composite purifying agent in Example 4. [Modes for carrying out the invention]
[0005] To further explain the method and effects achieved by the present invention, the following is described: The technical solutions of the present invention will be clearly and completely explained in conjunction with the experiments. In the following examples, PAEs are dimethyl phthalate (DMP) and diethyl phthalate (DE P), dibutyl phthalate (DBP), butyl benzyl phthalate (BBP), di( Six types of ethylhexyl phthalate (DEHP) and di-n-octyl phthalate (DOP) It contains EPA-grade control plasticizers, and all of the above reagents are analytically pure. Purchased from Lin Biochemical Technology Co., Ltd. Example 1: This example describes a bacterial colonization process for synergistically treating PAEs and Cd. A preparation method of a nickel composite purifying agent, wherein the bacterial colony carrier is polyvinyl alcohol- Adopt a combination of mercapto montmorillonite and reticulated carbon fiber, and include the following steps as follows: S1. Prepare functional bacterial colonies, S1-1. Domestication of Cd-resistant strains: On the premise of ensuring the normal growth of the strains and the functions of the strains that can decompose PAEs, induce the strains of the genera Gordonia, Erythrobacter, and Bacillus to be domesticated in the LB medium containing Cd 2 + S1-2. Activate the three strains after S1-1 treatment in LB medium for 24 h, and adjust OD 600 = 1. 0, S1-3. Mix the three strains after S1-2 treatment at a volume ratio of 1:1:1 to prepare functional bacterial colonies with PAEs decomposition and Cd resistance functions, The domestication results are shown in FIGS. 1 to 4. Here, FIG. 1 is a growth curve diagram of cadmium-resistant domesticated strains from 0 to 48 h, where Gordonia sp. belongs to the genus Gordonia, Rh odococcus sp. belongs to the genus Erythrobacter, and Bacillus sp. is the genus Bacillus. FIG. 2 is a diagram of the OD 2+ value when cadmium-resistant domesticated strains grow in LB medium with different Cd concentrations for 24 h. FIG. 3 is a diagram of the OD 600 value when cadmium-resistant domesticated strains grow in LB medium with different pH values for 24 h. FIG. 4 is a diagram of the degradation rates of six types of PAEs by cadmium resistant domesticated strains, 600 S2. Prepare a bacterial colony carrier, S2-1-B-1-1. First, dissolve montmorillonite in deionized water to form a suspension with a mass concentration of 4% Prepare the turbidity, stir at 50°C for 4 hours, then add PVA to deionized water to a mass concentration of 3%. Prepare a mixture of the following: Add solution S2-1-B-1-2 to the suspension in a volume ratio of 1:4, and follow the parameters below. Insert PVA into the montmorillonite gap: First, vibrate with ultrasound for 5 minutes and let stand for 1 hour, then vibrate with ultrasound for 5 minutes and let stand for 2 hours. Next, it is ultrasonically shaken for 5 minutes and left to stand for 10 hours, then stirred at 50°C for 3 hours, and finally sealed. After stopping the process, the material is subjected to 7d ultrasonic shaking at room temperature to obtain a colloidal suspension. The colloidal suspensions in S2-1-B-1-3 and S2-1-B-1-2 were separated by centrifugation. The separated material is then repeatedly washed with deionized water, and finally the separated material is mixed in a ratio of 1g:50mL. Dilute with deionized water and store as a reserve. S2-1-B-2, Low-temperature self-assembly: Processing of the colloidal suspension obtained in S2-1-B-1. After that, freeze at -10°C for 24 hours to expand the interlayer spacing and then extract the large interlayer montmorillonite. Gain, S2-1-B-3, Mercapto-modified: Large intercalated montmorillonite in S2-1-B-2 Add to an ethanol-aqueous solution of 3-mercaptopropyltrimethoxysilane and mix uniformly. After mixing, the temperature is controlled to 25°C and the pH to 10, and the reaction is carried out for 7 hours while continuing to stir. Large intercalated montmorillonite was prepared after modification with PT, and polyvinyl alcohol was used. It is written as Ru-mercaptomontmorillonite, The amount of large intercalated montmorillonite added is 1 g, and 3-mercaptopropyl trimethopropyl The amount of sicilan ethanol-aqueous solution added was 25 mL. S2-2, Preparation of lattice-like carbon fibers: First, 2 mm long lignocellulose is immersed in a 0.1 mol / L NaOH solution, and 550 Stir at r / min for 25 hours, then remove the lignocellulose and dry at 80°C. Finally, the material is calcined at a gradient temperature, and the calcined product is washed and dried to obtain a string-like carbon fiber. As a carbonized carrier, The temperature gradient is as follows: First, the temperature is raised from 20°C to 250°C at a rate of 5°C / min. Then, maintain the temperature for 6 minutes, and then raise the temperature from 250°C to 500°C at a rate of 22°C / min, 2 After keeping it warm for a while, it is finally cooled to 20°C in a furnace. S2-3, carrier composite: Polyvinyl alcohol-mercaptomontmorillony in S2-3-1 and S2-1-B-3 The biochar carrier in S2-2 is placed in pure water and mixed uniformly to obtain a mixture. The amount of polyvinyl alcohol-mercaptomontmorillonite added is 1g, and the biochar is The amount of body added was 4g, and the amount of pure water added was 110mL. The mixture in S2-3-2 and S2-3-1 was separated, and the separated product was washed and then 100 The material is dried at °C for 5 hours, and finally passed through a 200-mesh sieve to obtain a composite carrier, which is then used as a bacterial colony carrier. year, S3. Preparation of a fixed-type composite purifying agent. The functional bacterial colonies prepared in S3-1 and S1 and the bacterial colony carrier prepared in S2 were uniformly distributed. Mix them to obtain a mixed system. The amount of bacterial colony carrier added was 5g, and the amount of functional bacterial colonies added was 300mL. the law of nature, S3-2. First, the mixture obtained in S3-1 is subjected to a constant temperature of 30°C and light-shielded conditions at a rate of 200 r / min. The samples were then incubated with shaking for 24 hours, filtered and washed under sterile conditions, and then dried at 25°C. A bacterial colony complex purifying agent is prepared. Example 2: The description of this example is based on Example 1, but mainly describes a preparation method under different parameters. Explain, including the following steps: S1, Prepare functional bacterial colonies. S1-1, Domestication of Cd-resistant strains: Normal growth of strains and the function of strains that degrade PAEs. Under the condition that we guarantee it, strains of the genera Goldonella, Erythrobacter, and Bacillus are Cd 2 + Induced domestication using LB medium containing the active ingredient, The three strains treated with S1-2 and S1-1 were activated in LB medium for 24 hours, and OD 600 =1. Adjust to 0, Three strains treated with S1-2 were mixed in a volume ratio of 1:1:1 with S1-3, and PAE We prepared functional bacterial colonies that possess s-degradation and Cd-resistant functions. S2. Prepare the bacterial colony carrier. S2-1-B-1-1, First, montmorillonite is dissolved in deionized water to form a 2% mass concentration solution. Prepare the turbidity, stir at 50°C for 4 hours, then add PVA to deionized water to a mass concentration of 2%. Prepare a mixture of the following: Add solution S2-1-B-1-2 to the suspension in a volume ratio of 1:4, and follow the parameters below. Insert PVA into the montmorillonite gap: First, vibrate with ultrasound for 3 minutes and let stand for 1 hour, then vibrate with ultrasound for 4 minutes and let stand for 2 hours. Next, ultrasonic shaking for 5 minutes, then standing for 10 hours, followed by stirring at 45°C for 3 hours, and finally sealing. After stopping the process, the material is subjected to 5d ultrasonic shaking at room temperature to obtain a colloidal suspension. The colloidal suspensions in S2-1-B-1-3 and S2-1-B-1-2 were separated by centrifugation. The separated material is then repeatedly washed with deionized water, and finally the separated material is mixed in a ratio of 1g:50mL. Dilute with deionized water and store as a reserve. S2-1-B-2, Low-temperature self-assembly: Processing of the colloidal suspension obtained in S2-1-B-1. After that, freeze at -10°C for 24 hours to expand the interlayer spacing and then extract the large interlayer montmorillonite. Gain, S2-1-B-3, Mercapto-modified: Large intercalated montmorillonite in S2-1-B-2 Add to an ethanol-aqueous solution of 3-mercaptopropyltrimethoxysilane and mix uniformly. After mixing, the temperature is controlled to 25°C and the pH to 9, and the reaction is carried out for 6 hours while continuing to stir, and the mercapsules are formed. Large intercalated montmorillonite, modified by the agent, is prepared, and polyvinyl alcohol -Written as mercaptomontmorillonite, The amount of large intercalated montmorillonite added is 1 g, and 3-mercaptopropyl trimethopropyl The amount of sisirane ethanol-aqueous solution added was 20 mL. S2-2, Preparation of lattice-like carbon fibers: First, 1 mm long lignocellulose is immersed in a 0.1 mol / L NaOH solution, and 500 The mixture was stirred at r / min for 20 hours, and then the treated lignocellulose was separated and dried at 80°C. The material is then transferred to a different location, and finally fired at a gradient temperature. After firing, the product is washed and dried to obtain a string-like carbon fiber. As a biocarbon carrier, The temperature gradient is as follows: First, the temperature is raised from 20°C to 250°C at a rate of 4°C / min. Then, maintain the temperature for 5 minutes, and then raise the temperature from 250°C to 500°C at a rate of 20°C / min, 1 After keeping it warm for a while, it is finally cooled to 20°C in a furnace. S2-3, carrier composite: Polyvinyl alcohol-mercaptomontmorillony in S2-3-1 and S2-1-B-3 The biochar carrier in S2-2 is placed in pure water and mixed uniformly to obtain a mixture. The amount of polyvinyl alcohol-mercaptomontmorillonite added is 1g, and the biochar is The amount of body added was 3g, and the amount of pure water added was 100mL. The mixture in S2-3-2 and S2-3-1 was separated, and the separated product was washed and then 100 The material is dried at °C for 5 hours, and finally passed through a 200-mesh sieve to obtain a composite carrier, which is then used as a bacterial colony carrier. year, S3. Prepare a bacterial colony complex purifying agent. The functional bacterial colonies prepared in S3-1 and S1 and the bacterial colony carrier prepared in S2 were uniformly distributed. Mix them to obtain a mixed system. The amount of bacterial colony carrier added was 5g, and the amount of functional bacterial colonies added was 100mL. the law of nature, S3-2, First, the mixing system in S3-1 was mixed at 150 r / min, under constant temperature of 30°C and light-shielded conditions for 24 The samples were cultured by shaking, then filtered and washed under sterile conditions, and dried at 25°C to remove bacteria. Prepare a colony complex purifying agent. Example 3: This example describes a bacterial culture for synergistically treating PAEs and Cd. This is a method for preparing Ronnie's complex purifying agent, in which the bacterial colony carrier is sodium silicate- The combination of mercaptomontmorillonite and biocarbon particles is used, and the steps are as follows: It is: S1, Prepare functional bacterial colonies. S1-1, Domestication of Cd-resistant strains: Normal growth of strains and the function of strains that degrade PAEs. Under the condition that we guarantee it, strains of the genera Goldonella, Erythrobacter, and Bacillus are Cd 2 + Induced domestication using LB medium containing the active ingredient, The three strains treated with S1-2 and S1-1 were activated in LB medium for 24 hours, and OD 600 =1. Adjust to 0, S1-3, the three strains treated with S1-2 were mixed in a volume ratio of 1:1:1, and the PAEs were divided. We prepared functional bacterial colonies that possess both resorption and Cd resistance functions. S2. Prepare the bacterial colony carrier. S2-1-A, 10g montmorillonite, 0.6g sodium silicate, 1g 3-methyl 200 mL of 95% ethanol containing lucaptopropyltrimethoxysilane After adding it to the aqueous solution and mixing uniformly, the temperature is controlled to 25°C and the pH to 10, and stirring is continued. The reaction was carried out for 7 hours while mercapto was used to prepare porous montmorillonite after modification. And, it is written as sodium silicate-mercaptomontmorillonite, S2-2, Preparation of biochar particles: After crushing and polishing plant fibers, they are thermally decomposed under a nitrogen atmosphere. After decomposition, the product is polished and passed through a 200-mesh sieve to prepare biochar particles. As NT, The parameters for pyrolysis are as follows: The furnace temperature is raised to 600°C at a rate of 5°C / min. Then, hold for 2 hours, and after thermal decomposition, polish the biochar and pass it through a 200-mesh sieve to form biochar granules. Prepare the child, S2-3, carrier composite: Sodium silicate-mercaptomontmorillonite and S2 in S2-3-1 and S2-1-A -2 The biochar carrier is placed in pure water and mixed uniformly to obtain a mixture. The amount of sodium silicate-mercaptomontmorillonite added is 1g, and the biochar support The amount added was 4g, and the amount of pure water added was 110mL. The mixture in S2-3-2 and S2-3-1 was separated, and the separated product was washed and then 100 The material is dried at °C for 5 hours, and finally passed through a 200-mesh sieve to obtain a composite carrier, which is then used as a bacterial colony carrier. year, S3. Prepare a bacterial colony complex purifying agent. The functional bacterial colonies prepared in S3-1 and S1 and the bacterial colony carrier prepared in S2 were uniformly distributed. Mix them to obtain a mixed system. The amount of bacterial colony carrier added was 5g, and the amount of functional bacterial colonies added was 100mL. the law of nature, S3-2, First, the mixing system in S3-1 was mixed at 150 r / min, under constant temperature of 30°C and light-shielded conditions for 24 The samples were cultured by shaking, then filtered and washed under sterile conditions, and dried at 25°C to remove bacteria. Prepare a colony complex purifying agent. Example 4: The description of this example is based on Example 3, but mainly describes a preparation method under different parameters. Please explain in detail, including the following steps: S1, Prepare functional bacterial colonies. S1-1, Domestication of Cd-resistant strains: Normal growth of strains and the function of strains that degrade PAEs. Under the condition that we guarantee it, strains of the genera Goldonella, Erythrobacter, and Bacillus are Cd 2 + Induced domestication using LB medium containing the active ingredient, The three strains treated with S1-2 and S1-1 were activated in LB medium for 24 hours, and OD 600 =1. Adjust to 0, S1-3, the three strains treated with S1-2 were mixed in a volume ratio of 1:1:1, and the PAEs were divided. We prepared functional bacterial colonies that possess both resorption and Cd resistance functions. S2. Prepare the bacterial colony carrier. S2-1-A, 10g montmorillonite, 0.6g sodium silicate, 1g 3-methyl 200 mL of 95% ethanol containing lucaptopropyltrimethoxysilane After adding it to the aqueous solution and mixing uniformly, the temperature is controlled to 25°C and the pH to 9.5, and stirring is performed. Continue the reaction for 6 hours, and then examine the porous montmorillonite that has been denatured by mercapto. Prepared and labeled as sodium silicate-mercaptomontmorillonite, S2-2, Preparation of biochar particles: After crushing and polishing plant fibers, they are thermally decomposed under a nitrogen atmosphere. After decomposition, the product is polished and passed through a 200-mesh sieve to prepare biochar particles. As a carbonized carrier, The parameters for pyrolysis are as follows: The furnace temperature is raised to 600°C at a rate of 4°C / min. Then, hold for 2 hours, and after thermal decomposition, polish the biochar and pass it through a 200-mesh sieve. Prepare the particles, S2-3, carrier composite: Sodium silicate-mercaptomontmorillonite and S2 in S2-3-1 and S2-1-A -2 The biochar carrier is placed in pure water and mixed uniformly to obtain a mixture. The amount of sodium silicate-mercaptomontmorillonite added is 1g, and the biochar support The amount added was 3g, and the amount of pure water added was 100mL. The mixture in S2-3-2 and S2-3-1 was separated, and the separated product was washed and then 100 The material is dried at °C for 5 hours, and finally passed through a 200-mesh sieve to obtain a composite carrier, which is then used as a bacterial colony carrier. year, S3. Prepare a bacterial colony complex purifying agent. The functional bacterial colonies prepared in S3-1 and S1 and the bacterial colony carrier prepared in S2 were uniformly distributed. Mix them to obtain a mixed system. The amount of bacterial colony carrier added was 5g, and the amount of functional bacterial colonies added was 100mL. the law of nature, S3-2. First, the mixture obtained in S3-1 is prepared at 150 r / min at a constant temperature of 30°C under light-shielded conditions. The samples were then incubated with shaking for 24 hours, filtered and washed under sterile conditions, and then dried at 25°C. A bacterial colony complex purifying agent is prepared. Example 5: This example describes a bacterial colonization process for synergistically treating PAEs and Cd. This is a method for preparing a Knee complex purifying agent. This example is based on Example 4, but with modified conditions, and the following applies: They are the same except for the content: In S3-1: The amount of bacterial colony carrier added is 5g, and the amount of functional bacterial colonies added is 5 It is 0 mL. Example 6: This example describes a bacterial colonization process for synergistically treating PAEs and Cd. This is a method for preparing a Knee complex purifying agent. This example is based on Example 4, but with modified conditions, and the following applies: They are the same except for the content: In S3-1: The amount of bacterial colony carrier added is 5g, and the amount of functional bacterial colony added is 1 It is 50 mL. Example 7: This example describes a bacterial colonization process for synergistically treating PAEs and Cd. This is a method for preparing a Knee complex purifying agent. This example is based on Example 4, but with modified conditions, and the following applies: They are the same except for the content: S3-2: Maintain a constant temperature of 25°C. Example 8: This example describes a bacterial colonization process for synergistically treating PAEs and Cd. This is a method for preparing a Knee complex purifying agent. This example is based on Example 4, but with modified conditions, and the following applies: They are the same except for the content: During S3-2: Maintain a constant temperature of 35°C. Example 9: This example describes a bacterial colonization process for synergistically treating PAEs and Cd. This is a method for preparing a Knee complex purifying agent. This example is based on Example 4, but with modified conditions, and the following applies: They are the same except for the content: S3-2: Incubate for 48 hours with shaking under light-shielding conditions. Example 10: This example describes a bacterial culture for synergistically treating PAEs and Cd. This is a method for preparing Ronnie's complex purifying agent, and this example is based on Example 4 with modified conditions, as follows: It is the same except for the content: S3-2: Incubate for 72 hours with shaking under light-shielding conditions.
[0006] Comparative Example 1: The description of this example is for the synergistic treatment of bacterial colonies for PAEs and Cd. This is a method for preparing a Knee complex purifying agent. This example is based on Example 4, but with modified conditions, and the following applies: They are the same except for the content: Instead of functional bacterial colonies, we use the genus Goldonella. Comparative Example 2: The description of this example is for the synergistic treatment of bacterial colonies containing PAEs and Cd. This is a method for preparing a Knee complex purifying agent. This example is based on Example 4, but with modified conditions, and the following applies: They are the same except for the content: Instead of bacterial colony carriers, corn straw biochar, carbonized at 600°C, is used. Comparative Example 3: The description of this example is for the synergistic treatment of bacterial colonies with PAEs and Cd. This is a method for preparing a Knee complex purifying agent. This example is based on Example 4, but with modified conditions, and the following applies: They are the same except for the content: The bacterial colony carriers have not undergone mercaptochemical treatment. Test Example 1: This test example shows the Cd production by bacterial colony carriers in Example 4, Comparative Examples 2 and 3. 2+ of It is used to measure the saturated adsorption amount, and the specific steps are as follows: 0.1g A bacterial colony carrier is taken and 600 mg / L C containing 0.1 mol / L KNO3 is added. d 2+ Add to 20 mL of solution, maintain a solid-liquid ratio of 1 g:200 mL, and adjust the pH to 6.0. Then, shake at 25°C for 1 hour, let stand for 16 hours, centrifuge for 30 minutes, and collect the supernatant. CD 2+ The concentration was measured, and the test results are shown in Figure 5. As can be seen from the results: Cd by bacterial colony carrier in Example 4 2+ The saturation adsorption amount is, The result was significantly larger than comparative examples 2 and 3 (P<0.05), which is due to the composite prepared according to the present invention. The purifying agents include a composite purifying agent based on a biochar carrier, and a composite purifying agent that has not undergone mercaptotherapy. In comparison, heavy metal Cd 2+ The adsorption amount was significantly higher, and the passivation efficiency was clearly demonstrated. I showed it. Test Example 2: This test example concerns the inorganic salt system of the composite purifying agent in Examples 4-10 and Comparative Examples 1 and 2. It is used to measure the decomposition efficiency of PAEs, and the specific steps are as follows: Under sterile conditions, weigh 0.2g of the combined purifying agent and measure out six types of PAEs with a total concentration of 30mg / L. After adding it to 20 mL of inorganic salt medium and culturing for 5 days at 150 rpm under light-shielded conditions at 30°C, The ΣPAEs decomposition rate was measured and calculated, and the test results are shown in Figure 6. As can be seen from the results: The combined purifying agent in Example 4 showed a total decomposition rate of 86% for the six types of PAEs. The highest was 70%, as can be seen from the ΣPAEs decomposition rate of the composite purifying agents in Examples 5-10. The optimal preparation conditions for immobilized bacterial preparations are: bacterial ratio 1g:20mL, temperature 30°C, and 24 hours in the dark. The method involves shaking culture, and as can be seen from Comparative Example 1, functional bacterial colonies are functional Compared to single bacteria, the prepared complex purifier showed significantly improved ΣPAEs decomposition ability. Test Example 3: This test involved observing the composite purifying agent in Example 4 using a scanning electron microscope, and the test results were as follows: The result is shown in Figure 7. As can be seen from the results: Observation of the composite purifying agent in Example 4 using a SEM bioelectron microscope Furthermore, numerous functional bacterial strains are attached to its surface and pore structure, and the bacterial strains are aggregated. The compound purifier is attached to the composite material in a state or dispersed state, and the prepared composite purifier contains functional bacteria. This demonstrated that Ronnie can be efficiently concentrated and fixed. Test Example 4: This test example shows the decomposition efficiency of PAEs in contaminated soil by the composite remediation agent in Example 4. It is also used to measure the passivation efficiency of heavy metals such as Cd. The composite remediation agent in Example 4 was applied to soil contaminated with PAEs and heavy metal Cd at a concentration of 1%. After culturing in the dark for 50 days, the PAEs content and heavy metal fugue morphology were measured, and the test results are shown in Figure 8. This is shown in Figure 9. Conclusion: As can be seen from Figure 8, when the combined remediation agent in Example 4 is applied to the combined contaminated soil, The ΣPAEs content decreased from 2.56 mg / kg to 1.17 mg / kg, and the degradation rate was 54%. The PAEs degradation efficiency is 0.14%, and as can be seen in Figure 9, the combined purifying agent is suitable. The Cd content in the treated soil under the exchange state decreased by 37.06%, and the carbonate binding state and iron manganese were also reduced. The oxide bond state and residual state Cd were 19.26%, 13.00%, and 12.5%, respectively. It increased by 5%, which is because the application of the combined remediation agent causes the soil's cadmium to be in a more active exchange state. The conversion to a stable carbonate bond state, an iron-manganese oxide bond state, and a residual state is promoted, It demonstrated that it possesses high passivation efficiency for heavy metals.
Claims
1. S1, a step of preparing functional bacterial colonies, S1-1, Domestication of Cd-resistant strains: Goldonella, Erythrobacter, and Bacillus Strains of the genus with gradient concentration Cd ２＋ The process involves inducing domestication using LB medium, The aforementioned strain of the genus Goldonella is Goldonia sp., and the genus Erythrobacter The strain is Rhodococcus sp., and the strain of the genus Bacillus is Bacillus. sp. The three bacterial strains treated with S1-2 and S1-1 were activated in LB medium for 24 hours, and then OD ６００ = 1 The process of adjusting to 0, S1-3, the three bacterial strains treated with S1-2 were mixed in a volume ratio of 1:1:1, and PAEs The process includes a step of preparing functional bacterial colonies that have degradation and Cd resistance functions, S2, a step of preparing a bacterial colony carrier, S2-1, the process of preparing mercaptomontmorillonite, S2-2, A step to prepare a biochar carrier, S2-3, Carrier composite: First, mercaptomontmorillonite in S2-1 and the ba in S2-2 Iocarbon carriers are placed in pure water, mixed uniformly, then centrifuged, washed, and dried to remove bacterial colonies. The process includes obtaining a carrier, S3, a step of preparing a bacterial colony complex purifying agent, The functional bacterial colonies prepared in S3-1 and S1 and the bacterial colony carrier prepared in S2 were mixed together. The first step is to mix the ingredients and then obtain the mixed system. Let n be the multiplier coefficient, where n ∈ R ＋ Therefore, if the amount of bacterial colony carrier added is 1 ng Yes, the amount of functional bacterial colonies added was 20 nmL. S3-2, First, the mixed system obtained in S3-1 is heated at a constant temperature of 30°C at a rate of 150-200 r / min. The samples were cultured for 24 hours with shaking under light-shielding conditions, then filtered, washed, and incubated at 25°C under sterile conditions. The process includes a step of transferring the material to dry and preparing a bacterial colony complex purifying agent. A bacterial colony complex for synergistically treating PAEs and Cd, characterized by containing a bacterial colony complex Method for preparing a purifying agent.
2. In S2-1, the mercaptomon morillonite is sodium silicate-mercaptomon It is molylonite or polyvinyl alcohol-mercaptomon molylonite. A bacterial colony for synergistically treating PAEs and Cd as described in claim 1. Method for preparing a complex purifying agent.
3. The method for preparing the sodium silicate-mercaptomontmorillonite is as follows: S2-1-A, montmorillonite, sodium silicate, 3-mercaptopropyl trimeth Xysilane is added to an ethanol-aqueous solution and mixed uniformly, then the temperature is raised to 25°C and the pH to 9. Control the temperature to 5-10 and continue stirring while reacting for 6-7 hours until the mixture is denatured by mercapto. After preparing the small-pore montmorillonite, sodium silicate-mercaptomontmorillonite was used. He wrote, Here, montmorillonite, sodium silicate and 3-mercaptopropyl trimethohydrate The mass ratio of sisirane is 1:0.06:0.
1. The mass concentration of the ethanol-aqueous solution is 95%. The mass sum of montmorillonite and sodium silicate is M. １ , 3-mercaptopropyl trimeth The sum of the volumes of xysilane and ethanol aqueous solutions is V. １ Therefore, M １ : V １ = 1.06g:2 The PAEs and Cd according to claim 2, characterized in that the amount is 0 mL, A method for preparing a bacterial colony complex purifying agent.
4. The method for preparing the polyvinyl alcohol-mercaptomontmorillonite is as follows: ： S2-1-B-1, Interlayer insertion of organic matter: First, montmorillonite is dissolved in deionized water and the Prepare a suspension with a mass concentration of 2-4%, then add PVA to deionized water to a mass concentration of 2-3%. Prepare a % mixture, then add the solution to the suspension in a volume ratio of 1:4, and apply 5-7 d ultrasonic waves. Wave vibration is performed to obtain a colloidal suspension. S2-1-B-2, Low-temperature self-assembly: The colloidal suspension obtained in S2-1-B-1 is -1 After freezing at 0°C for 24 hours and expanding the interlayer spacing, large interlayer montmorillonite was obtained. S2-1-B-3, Mercapto-modified: Large intercalated montmorillonite in S2-1-B-2 Add to an ethanol-aqueous solution of 3-mercaptopropyltrimethoxysilane and mix uniformly. After mixing, control the temperature to 25°C and the pH to 9.5-10, and continue stirring while reacting for 6-7 hours. This process yields large interlayer montmorillonite modified by mercapto, and polyvinyl Alcohol-mercaptomontmorillonite, Here, large intercalated montmorillonite and 3-mercaptopropyltrimethoxysilane The mass ratio is 1:0.1, The mass concentration of the ethanol-aqueous solution is 95%. The mass of a large interlayer montmorillonite is M ２ , 3-mercaptopropyltrimethoxysilane , let the volume sum of the ethanol - aqueous solution be V ２ Then, M ２ : V ２ = 1 g : 20 mL, this A bacterial colonization method for synergistically treating PAEs and Cd according to claim 2, characterized by the above. Method for preparing a complex purifying agent.
5. The method for interlayer insertion of organic matter in S2-1-B-1 is as follows: S2-1-B-1-1, First, dissolve montmorillonite in deionized water to a mass concentration of 2-4 Prepare a % suspension, stir at 45-50°C for 3-4 hours, then add PVA to deionized water. Prepare a mixed solution with a mass concentration of 2-3%, S2-1-B-1-2, the above solution is added to the suspension in a volume ratio of 1:4, and according to the following parameters Then insert PVA into the gaps of the montmorillonite: First, vibrate with ultrasound for 3-5 minutes and let stand for 1 hour, then vibrate with ultrasound for 4-5 minutes. Let it stand for 10 hours, then soak it with ultrasound for 5 minutes and let it stand for 10 hours, then stir at 45-50°C for 3 hours. Mix the mixture, then seal it and shake it ultrasonically at room temperature for 5-7 days to obtain a colloidal suspension. The colloidal suspensions in S2-1-B-1-3 and S2-1-B-1-2 were separated by centrifugation. The separated material is then repeatedly washed with deionized water, and finally the separated material is diluted with deionized water. The PAEs and Cd described in 4 are compounded by releasing and storing them for reserve. A method for preparing a bacterial colony complex purifying agent for targeted treatment.
6. The biochar carrier paired with the aforementioned sodium silicate-mercaptomontmorillonite is Biochar particles, which are paired with the aforementioned polyvinyl alcohol-mercaptomontmorillonite. The PA according to claim 2, characterized in that the biochar carrier to be attached is a string-like carbon fiber. A method for preparing a bacterial colony complex purifying agent for synergistically treating Es and Cd.
7. The method for preparing the biochar particles is as follows: plant fibers are crushed and polished, and then inactivated. The product is thermally decomposed under a thermal atmosphere, and after thermal decomposition, the product is polished and passed through a 200-mesh sieve. Prepare carbon particles, The parameters for the aforementioned pyrolysis are as follows: the furnace temperature is increased from room temperature at a rate of 4-5°C / min. The PAEs described in 6 are characterized by raising the temperature to 600°C and holding it there for 2 hours. A method for preparing a bacterial colony complex purifying agent for synergistically treating Cd.
8. The method for preparing the aforementioned lattice-like carbon fibers is as follows: First, 1-2 mm long lignocellulose is immersed in a 0.1 mol / L NaOH solution, 5 After stirring at 00-550 r / min for 20-25 hours, remove the lignocellulose and 80 The material is then dried at °C, and finally calcined at a gradient temperature. The calcined product is then washed and dried to obtain stringy carbon. Obtaining fibers, The aforementioned temperature gradient is as follows: First, at a rate of 4-5°C / min, from 20°C to 250°C The temperature is raised, maintained for 5-6 minutes, and then heated from 250°C to 50°C at a rate of 20-22°C / min. The process involves raising the temperature to 0°C, maintaining the temperature for 1-2 hours, and finally cooling it to 20°C in a furnace. A bacterial colony complex purifying agent for synergistically treating PAEs and Cd as described in item 6. Preparation method.
9. The carrier composite method in S2-3 is as follows: S2-3-1, mercaptomontmorillonite in S2-1 and biochar carrier in S2-2 Dissolve in pure water and mix uniformly to obtain a mixture, with n as the multiplication coefficient, where n ∈ R ＋ Therefore, the above M The amount of lucaptomontmorillonite added was 1 ng, and the amount of biochar support added was 3 ng. The amount of pure water added is 100 nmL. The mixture obtained in S2-3-2 and S2-3-1 is separated, and the separated product is washed to form 1 The process involves drying at 0°C for 5 hours, followed by passing the material through a 200-mesh sieve to obtain a composite carrier. A bacterial colony complex for synergistically treating PAEs and Cd according to claim 1. Method for preparing a purifying agent.