Method for removing polycyclic aromatic hydrocarbons from soil using recyclable laccase system

Abstract

To provide a method for removing polycyclic aromatic hydrocarbons from soil using a recyclable laccase system.SOLUTION: The method includes: a step S1 of preparing a polyaniline-BiVO4 / ZnO fiber membrane metal kernel laccase; a step S2 of preparing a surfactant-based laccase; a step S3 of applying the polyaniline-BiVO4 / ZnO fiber membrane metal kernel laccase and the surfactant-based laccase to the soil; and a step S4 of recovering the polyaniline-BiVO4 / ZnO fiber membrane metal kernel laccase. By co-applying the improved polyaniline-BiVO4 / ZnO fiber membrane metal kernel laccase and the surfactant-based laccase to soil contaminated with polycyclic aromatic hydrocarbons, the polycyclic aromatic hydrocarbons in the soil can be efficiently removed.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to the technical field of soil pollution management, in particular to the recyclable laccase The present invention relates to a method for removing polycyclic aromatic hydrocarbons from soil using a system. [Background technology]

[0002] Polycyclic aromatic hydrocarbons (PAHs) are aromatic hydrocarbons that contain two or more benzene rings. PAHs are known to cause respiratory irritation due to their toxicity, genotoxicity, mutagenicity and carcinogenicity. Major effects on the human body include damage to the organs, circulatory system, nervous system, liver, and kidneys. It is recognized as an organic pollutant. Removing polycyclic aromatic hydrocarbons (PAHs) from the environment will restore polluted environments. These are considered to be the most important methods for reducing CO2 emissions. These include incineration, alkaline catalyzed dechlorination, UV oxidation, Many physical and chemical treatment methods, such as immobilization and solvent extraction, have been attempted. These traditional methods have the disadvantage of being costly, complex and difficult to regulate. In many cases, environmental purification technologies are unable to completely remove pollutants, and simply reduce the amount of pollutants. They simply move it from one environment to another or form other pollutants. Bioremediation and semiconductor photocatalytic decomposition technologies remove polycyclic aromatic hydrocarbons It is a better way to Recent studies have shown that laccase is effective in managing PAHs. Further research is needed to rationally utilize caseins for the purification of PAHs. Summary of the Invention

[0003] In response to the above-mentioned problems, the present invention provides a recyclable laccase system that can be used to treat soil. A method for removing polycyclic aromatic hydrocarbons from soil is provided. The technical solutions of the present invention are as follows: Removal of polycyclic aromatic hydrocarbons from soil by a recyclable laccase system The law is S1, Preparation of polyaniline-BiVO4 / ZnO fibrous membrane metallokernel laccase: parts by weight 1-2 parts of BiVO4 solid powder, 5-6 parts of (CH3COO)2Zn solid powder, NaOH Add 4 parts of the solid powder to 15-20 parts of 75% ethanol solution and store at room temperature for 30 min. n A mixture was obtained by magnetic stirring, and the mixture was mixed with the polyaniline fiber membrane fragments in a ratio of 15 to 25:1. The mixture was mixed in a weight ratio and shaken for 1-2 hours. After that, the polyaniline fiber membrane fragments were taken out and Incubate in a flask for 2-3 hours, then mix with the above mixture and incubate the polyaniline. The phosphorus fiber membrane fragments were mixed in a weight ratio of 10–15:1, heated in a water bath for 3–4 h, and polyaniline was added. The polyaniline fiber membrane fragments were taken out and dried to obtain the activated polyaniline fiber membrane fragments. The activated polyaniline fiber membrane fragments and the laccase solution were mixed and stirred in a weight ratio of The mass concentration of the Zeolite solution was set to 5-20 mg / mL, and the polyaniline-BiVO4 / ZnO fibrous membrane was Obtaining a metallokernel laccase; S2. Preparation of surfactant-based laccase: rhamnolipid, laccase solution and deionized water. The resulting surfactant-based laccase was mixed with water. The mass concentration of rhamnolipid in the case of lacquer is 300 to 500 mg / L. A step of adjusting the mass concentration of the Ze solution to 100 to 200 mg / L; S3, Polyaniline-BiVO4 / ZnO fibrous membrane metal kernel laccase to soil and interface Application of laccase based surfactant: Metal kernel lacquer to soil at weight ratio of 3:5-10 Laccase and surfactant-based laccase are applied at a depth of 20-80 cm. Including, In one aspect of the present invention, the method includes a step of recovering and reusing the metallokernel laccase. It further includes the S4 S4, Metal kernel laccase recovery cycle: The laccase applied in step S3 through the application device The polyaniline-BiVO4 / ZnO fibrous membrane was used to recover and recover the metallokernel laccase. The polyaniline-BiVO4 / ZnO fibrous membrane metallokernel laccase was washed with deionized water. After that, it was applied to the soil in step S3 again, and the recovered polyaniline-BiVO4 / ZnO fibers were The membrane metallokernel laccase and surfactant-based laccase were added to soil at a weight ratio of 4:7. Apply. In one aspect of the present invention, the size of the polyaniline fiber membrane fragments in step S1 is 0 .5mm~5mm, and the thickness of the polyaniline fiber membrane fragments is 0.5μm~2mm; The shaking speed of the mixing shaker was 1000 rpm, and the incubation temperature in the oven was 4 The temperature of the water bath was 100°C, and the polyaniline fiber membrane fragments after activation were The mixing and stirring speed of the laccase solution was 400 rpm. In one aspect of the present invention, the method for preparing the BiVO4 solid powder in step S1 is as follows: As follows: Take 0.01 mol of Bi(NO3)3·5H2O and add 20 mL of ethyl acetate. The mixture was dissolved in ethylene glycol and stirred for 60 min to obtain mixture A. Take NH4VO3 and dissolve it in 20mL of deionized water, and stir the mixture for 60min. After obtaining B, add mixed solution B dropwise to mixed solution A and stir magnetically for 60 minutes. Then add it to the reactor and stir for 15 minutes. The reaction was carried out at 0°C for 10 hours, and then cooled to room temperature in a furnace. The product was then centrifuged in a centrifuge. The BiVO4 solid was obtained and dried in vacuum at 75 °C for 2 h. Description: The BiVO4 solid prepared by this method has good stability as a laccase carrier. , low cost and increases the loading and activity of laccase. As one aspect of the present invention, the polyaniline-BiVO4 / ZnO fiber in step S3 The application sequence of the membrane metallokernel laccase and surfactant-based laccase was as follows: First, surfactant-based laccase is applied to the soil by layer infiltration, and then Polyaniline-BiVO4 / ZnO fibrous membrane with metallokernel laccase layered on soil The permeation rate of surfactant-based laccase was 0.3-0.6 L / h. The penetration rate is 2 to 3 L / m 2 and the spacing between the permeation layers of each surfactant-based laccase The permeation layer of each surfactant-based laccase and each polyaniline- The BiVO4 / ZnO fiber membrane metallokernel laccase laying layer had a one-to-one correspondence. Description: Optimizing the application sequence of metallokernel laccase and surfactant-based laccase By infiltrating the soil with surfactant-based laccase, the metal kernel laccase Application of laccase increased the activity of metal kernel laccase and reduced the amount of polycyclic aromatic hydrocarbons in the soil. It is possible to improve the treatment efficiency of aromatic hydrocarbons and simultaneously recover metal kernel laccase. . In one aspect of the present invention, in step S3, polyaniline-BiVO 4 / ZnO fiber membrane metallokernel laccase and surfactant-based laccase were applied, and The application device includes a cartridge disposed in a hollow space, the cartridge containing polyaniline. Multiple disks for distributing phosphorus-BiVO4 / ZnO fibrous membrane metallokernel laccase a dispenser assembly for dispensing the cartridge in an axial direction; Distributed in a spiral shape along the The dispenser assembly is disposed inside an outer slider and the outer slider. an inner slider slidably connected to said cartridge; and a front end of said outer slider The rear end of the outer slider is slidably connected to a first slot provided in a side wall of the flange. Extending to the inner wall of the cartridge, there are fixed blocks on both the upper and lower sides of the rear end of the outer slider. The front end of the fixed block is in contact with the front end inner wall of the cartridge via a first spring. The rear end of the fixed block is connected to the rear end inner wall of the cartridge via a telescopic rod. The front end of the inner slider fits into a second slot on the front wall of the outer slider. The rear end of the inner slider is slidably connected to the outer slider. abutting a rear end of the cavity, the rear end of the cavity being provided with a third slot, A porous baffle is provided at the tip of the third slot, and the rear end of the inner slider is supported by the second spring. and connected to the center of the porous baffle. A surfactant is attached to the inner wall of the cartridge corresponding to the tip of each of the dispenser assemblies. Each of the soils has an infiltration hole for infiltrating the laccase-based activator into the soil. The upper and lower sides of the inner slider are covered with polyaniline-BiVO4 / ZnO fiber membrane metal carburettes. A groove for disposing the nerlaccase is provided, and a permeation membrane is provided on the surface of the groove at the bottom. A magnetic attraction disk is provided between the two permeable membranes, and the upper and lower rear ends of the inner slider An engagement block is provided on each side, and the engagement block is an inner part of the outer slider. The cartridge is slidably connected to the sliding grooves provided on both the upper and lower sides of the cavity, and the top of the cartridge is provided with an air hole. An inlet and a liquid inlet are provided, and a drill bit is provided at the bottom of the cartridge. Description: By installing the application device, polyaniline-BiVO4 / ZnO fiber membrane metal carnet The application of laccase and surfactant-based laccase has been completed, and the method of the present invention is expected to be realized. By providing a groove, the polyaniline-BiVO4 / ZnO fiber membrane metal kernel la Carboxyase and recovered polyaniline-BiVO4 / ZnO fibrous membrane metal kernel lacquer The polyaniline-BiVO4 / ZnO fiber membrane can be simultaneously placed on the metal kernel. It is convenient for recovering laccase. Preferably, when the telescopic rod is compressed to the shortest position, the front end of the outer slider is When the second spring is fully extended, the groove is fully extended and the first slot is inserted. Located outside the second slot, A third spring is provided on both the upper and lower sides of the penetration hole, and a penetration hole is provided on the tip of each of the two third springs. The restrictive baffle for blocking the through hole is connected together, and when the third spring naturally expands, The limiting baffle fits onto the rear end of the inner slider and limits the number of dispenser assemblies. The number of holes is 5 to 6, and the total infiltration rate of all the infiltration holes is 0.3 to 0.6 L / h. Description: By optimizing and limiting the length of the telescopic rod, polyaniline-BiVO4 / Z The best treatment effect of nO fiber membrane metal kernel laccase was achieved, and at the same time the operation of the equipment was made smooth. The restriction baffle can be installed to reduce the surfactant base. At the same time, the laccase in the gas mixture is prevented from entering the cavity through the porous baffle. The holes can be sealed. Effect of the Invention

[0004] The present invention has the following beneficial effects: (1) The recyclable laccase system of the present invention decomposes polycyclic aromatic hydrocarbons in soil The method for removing the fluorine from the improved polyaniline-BiVO4 / ZnO fibrous membrane metal kernel Co-application of laccase and surfactant-based laccase to polycyclic aromatic hydrocarbon-contaminated soils This allows efficient removal of polycyclic aromatic hydrocarbons from the soil, optimizing the application method. By optimizing the soil, the surfactant-based laccase is first introduced into the soil by layer infiltration. After application, the polyaniline-BiVO4 / ZnO fiber membrane was covered with a layer of metal kernel laccase. The polyaniline-BiVO4 / ZnO fiber membrane was applied to the soil as shown in Fig. 1. This prolongs the activity of glycerol, improving the effectiveness of treating polycyclic aromatic hydrocarbons in soil. At the same time, the polyaniline-BiVO4 / ZnO fibrous membrane facilitates the recovery of metallokernel laccase. It can be. (2) The recyclable laccase system of the present invention decomposes polycyclic aromatic hydrocarbons in soil. Polyaniline-BiVO4 / ZnO fibrous membrane solids prepared by the method of removing oxygen were It has good stability as a laccase carrier, is inexpensive, and has high loading and activity of laccase. and co-applied with surfactant-based laccase to polycyclic aromatic hydrocarbon-contaminated soil. By using this method, the removal rate of polycyclic aromatic hydrocarbons in soil reached 35-50%, and the recovered polycyclic aromatic hydrocarbons were Rianiline-BiVO4 / ZnO fibrous membrane metallokernel laccase and newly prepared polyaniline Optimization and tailoring of Nilin-BiVO4 / ZnO fibrous membrane metallokernel laccase and its mixing By using polyaniline-BiVO4 / ZnO fiber membrane metal kernel lacquer Further improve the efficiency of the recovery and utilization of the enzyme, and avoid a decline in the treatment effect under the assumption of a high recovery and utilization rate. It is environmentally friendly, efficient and suitable for further expansion. (3) The recyclable laccase system of the present invention decomposes polycyclic aromatic hydrocarbons in soil. The method of removing the dye is to install an application device and apply it to the polyaniline-BiVO4 / ZnO fiber. The application of membrane metallokernel laccase and surfactant-based laccase was completed, and the present invention The method is most suitable for realizing the grooved polyaniline-BiVO4 / ZnO fiber membrane. Metallokernel laccase and the recovered polyaniline-BiVO4 / ZnO fibrous membrane metallokernel The laccase can be placed on the polyaniline-BiVO4 / ZnO fiber at the same time. It is convenient for the recovery of membrane metallokernel laccase and optimizes and limits the length of the telescopic rod. The best treatment of metallokernel laccase was achieved by polyaniline-BiVO4 / ZnO fibrous membrane The effect is achieved, and at the same time, the operation of the equipment can be made smoother, the failure rate can be reduced, and the control By providing a limiting baffle, the surfactant-based laccase can enter the cavity through the porous baffle. At the same time, the penetration holes can be sealed after gas injection to prevent the gas from entering the inlet. [Brief description of the drawings]

[0005] [Figure 1] FIG. 15 is a structural schematic diagram of the application device of Example 12 of the present invention. [Diagram 2] FIG. 15 is a structural schematic diagram of the dispenser assembly inside the application device of Example 12 of the present invention. [Diagram 3] FIG. 23 is a structural schematic diagram of the dispenser assembly inside the application device of Example 12 of the present invention during dispensing. [Figure 4]FIG. 15 is a structural schematic diagram of the porous baffle of the application device of Example 12 of the present invention. [Diagram 5] FIG. 2 is a structural schematic diagram of the application device of Example 12 of the present invention during discharging.

[0006] [Explanation of symbols] 1 Cartridge 11 First Slot 12 Penetration holes 13 Air inlet 14 Liquid inlet 15 Drill Bits 2 Outer slider 21 Fixed Block 22 First Spring 23 Telescopic Rod 24 2nd Slot 25 Cavity 26 3rd Slot 27 Porous Baffle 28 Sliding groove 3 Inner slider 31 Second Spring 32 Placement groove 33 Magnetic attraction disk 34 Engagement block 4 Restriction Baffle 41 Third Spring DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0007] Example 1 Removal of polycyclic aromatic hydrocarbons from soil by a recyclable laccase system The method includes the steps of: S1, Preparation of polyaniline-BiVO4 / ZnO fibrous membrane metallokernel laccase: parts by weight 1 part BiVO4 solid powder, 5 parts (CH3COO)2Zn solid powder, 1 part NaOH solid powder Add 4 parts of the mixture to 17 parts of 75% ethanol solution and stir with a magnetic stirrer at room temperature for 30 min. A mixture was obtained, and the mixture was mixed with the polyaniline fiber membrane fragments in a weight ratio of 15 to 25:1. After mixing and shaking for 1 h, the polyaniline fiber membrane fragments were taken out and placed in an oven for 2 h. The polyaniline fiber membrane fragments were then incubated with the mixture for 12: The mixture was mixed in a weight ratio of 1 and heated in a water bath for 3 h. The polyaniline fiber membrane fragments were then removed and dried. The activated polyaniline fiber membrane fragments were obtained by mixing the activated polyaniline fiber fragments with the activated polyaniline fiber fragments at a weight ratio of 5:1. The fiber membrane fragments and the laccase solution were mixed and stirred until the mass concentration of the laccase solution was 12 mg / mL. The polyaniline-BiVO4 / ZnO fibrous membrane metallokernel laccase was obtained. The size of the polyaniline fiber membrane fragments was 1 mm, and the average thickness of the polyaniline fiber membrane fragments was 50 μm. The rotating speed of the mixer was 1000 rpm, and the incubation time was 1 h. The heating temperature was 400°C, and the water bath heating temperature was 100°C. The polyaniline fibers after activation The mixing and stirring speed of the membrane fragments and laccase solution was 400 rpm. The chemical reaction between (CH3COO)2Zn and NaOH produces zinc hydroxide, which is When heated to temperatures of 300-400 degrees, it decomposes to produce ZnO. The preparation method of BiVO4 solid powder is as follows: 0.01 mol Bi(NO3) Dissolve 3·5H2O in 20 mL of ethylene glycol solution and mix with magnetic stirring for 60 min. Mixture A was obtained, and 0.01 mol of NH4VO3 was dissolved in 20 mL of deionized water and cooled for 60 min. After obtaining mixed solution B by magnetic stirring, add mixed solution B dropwise to mixed solution A and magnetically stir for 60 min. The mixture was then placed in a reactor and reacted at 150°C for 10 hours. The mixture was then cooled to room temperature in an oven and the product was centrifuged. The BiVO4 solid was obtained by centrifuging in a centrifuge and then dried in vacuum at 75℃ for 2h. S2. Preparation of surfactant-based laccase: rhamnolipid, laccase solution and deionized water. The resulting surfactant-based laccase was mixed with water. The mass concentration of rhamnolipid in the laccase solution was 400 mg / L. The mass concentration is 150 mg / L. S3, Polyaniline-BiVO4 / ZnO fibrous membrane metal kernel laccase to soil and interface Application of activator-based laccase: Metal kernel laccase was applied to soil at a weight ratio of 3:7. Laccase based surfactant was applied at a depth of 60cm. Polyaniline-BiVO4 / ZnO fibrous membrane metallokernel laccase and boundary during step S3 The application sequence of surfactant-based laccase is as follows: first, surfactant-based After laccase was applied to the soil by layer infiltration, polyaniline-BiVO4 / Zn O Fiber membrane metallokernel laccase was applied to the soil as a layer, and surfactant-based The permeation rate of laccase in the water was 0.5 L / h, and the permeation volume was 2.5 L / m 2 And in various fields The spacing between the surfactant-based laccase permeation layers was 12 cm. Permeation layer of laccase and metal kernel lacquer on polyaniline-BiVO4 / ZnO fibrous membranes The layers of Ze correspond one-to-one.

[0008] Example 2 This example differs from Example 1 in the following respects: the mass concentration of the laccase solution is different. S2. Preparation of surfactant-based laccase: rhamnolipid, laccase solution and deionized water. The resulting surfactant-based laccase was mixed with water. The mass concentration of rhamnolipid in the laccase solution was 400 mg / L. The mass concentration is 100 mg / L. Example 3 This example differs from Example 1 in the following respects: the mass concentration of the laccase solution is different. S2. Preparation of surfactant-based laccase: rhamnolipid, laccase solution and deionized water. The resulting surfactant-based laccase was mixed with water. The mass concentration of rhamnolipid in the laccase solution was 400 mg / L. The mass concentration is 200 mg / L. Example 4 This embodiment differs from embodiment 1 in the following respects: S3, Polyaniline-BiVO4 / ZnO fibrous membrane metal kernel laccase to soil and interface Application of laccase based on surfactant: Polyaniline-BiVO4 / in soil at a weight ratio of 3:5. ZnO fiber membrane metallokernel laccase and surfactant-based laccase were applied, and the application depth The degree is 60 cm. Example 5 This embodiment differs from embodiment 1 in the following respects: S3, Polyaniline-BiVO4 / ZnO fibrous membrane metal kernel laccase to soil and interface Application of laccase based on surfactant: Polyaniline-BiVO4 / in soil at a weight ratio of 3:9. ZnO fiber membrane metallokernel laccase and surfactant-based laccase were applied, and the application depth The degree is 60 cm. Example 6 This embodiment differs from embodiment 1 in the following respects: S3, Polyaniline-BiVO4 / ZnO fibrous membrane metal kernel laccase to soil and interface Application of laccase based surfactant: Polyaniline-BiVO4 in soil at a weight ratio of 3:10 / ZnO fiber membrane metallokernel laccase and surfactant-based laccase were applied and applied The depth is 60cm. Example 7 This embodiment differs from embodiment 1 in the following respects: S3, Polyaniline-BiVO4 / ZnO fibrous membrane metal kernel laccase to soil and interface Application of activator-based laccase: Polyaniline-BiVO4 in soil at a weight ratio of 3:12 / ZnO fiber membrane metallokernel laccase and surfactant-based laccase were applied and applied The depth is 60cm. Example 8 This embodiment differs from embodiment 1 in the following respects: S3, Polyaniline-BiVO4 / ZnO fibrous membrane metal kernel laccase to soil and interface Application of laccase based on surfactant: Polyaniline-BiVO4 / in soil at a weight ratio of 3:4. ZnO fiber membrane metallokernel laccase and surfactant-based laccase were applied, and the application depth The degree is 60 cm. Example 9 This embodiment differs from the first embodiment in the following respects, and further includes step S4: S4. Recovery of metallokernel laccase from polyaniline-BiVO4 / ZnO fibrous membrane: Application The polyaniline-BiVO4 / ZnO fiber membrane metallization applied in step S3 through the Lulaccase was harvested and recovered from polyaniline-BiVO4 / ZnO fibrous membrane metal kernel The laccase was washed with deionized water and then applied to the soil in step S3 again. Aniline-BiVO4 / ZnO fibrous membrane metallokernel laccase and surfactant-based laccase Case is applied to the soil in a weight ratio of 4:7. Example 10 This embodiment differs from the first embodiment in the following respects: Some specific parameters of steps S1 and S3 The data selection is different. S1, Preparation of polyaniline-BiVO4 / ZnO fibrous membrane metallokernel laccase: parts by weight 1 part BiVO4 solid powder, 5 parts (CH3COO)2Zn solid powder, 1 part NaOH solid powder Add 4 parts of the mixture to 15 parts of 75% ethanol solution and stir magnetically at room temperature for 30 min. A mixture was obtained, and the mixture was mixed with the polyaniline fiber membrane fragments in a weight ratio of 15:1 and then left for 1 h. Mix and shake, then take out the polyaniline fiber membrane fragments and place them in an oven for 2 h incubation. The polyaniline fiber membrane fragments were then incubated with the mixture in a 10:1 ratio. The mixture was mixed in the ratio of 0.1 to 0.25, heated in a water bath for 3 h, and the polyaniline fiber membrane fragments were taken out and dried to confirm the activity. The activated polyaniline fiber membrane fragments were obtained and then treated with 5:1 weight ratio. The debris and the laccase solution were mixed and stirred to a mass concentration of 5 mg / mL. Polyaniline-BiVO4 / ZnO fibrous membrane for metallokernel laccase synthesis The size of the fragments was 0.5 mm, and the average thickness of the polyaniline fiber membrane fragments was 0.5 μm. The rotation speed of the mixer was 1000 rpm, and the incubation temperature in the oven was The temperature of the water bath was 400°C, and the water bath heating temperature was 100°C. The polyaniline fiber membrane fragments after activation The mixing and stirring speed of the laccase solution was 400 rpm. Polyaniline-BiVO4 / ZnO fibrous membrane metallokernel laccase and boundary during step S3 The application sequence of surfactant-based laccase is as follows: first, surfactant-based After laccase was applied to the soil by layer infiltration, polyaniline-BiVO4 / Zn O Fiber membrane metallokernel laccase was applied to the soil as a layer, and surfactant-based The permeation rate of laccase in the water was 0.3 L / h, and the permeation volume was 2 L / m 2 Each interface activity The spacing between the permeation layers of each surfactant-based laccase was 10 cm. The permeation layer of laccase and the polyaniline-BiVO4 / ZnO fibrous membranes containing metal kernel laccase The laying layers correspond one-to-one. Example 11 This embodiment differs from the first embodiment in the following respects: Some specific parameters of steps S1 and S3 The data selection is different. S1, Preparation of polyaniline-BiVO4 / ZnO fibrous membrane metallokernel laccase: parts by weight 2 parts of BiVO4 solid powder, 6 parts of (CH3COO)2Zn solid powder, and NaOH solid powder Add 4 parts of the mixture to 20 parts of 75% ethanol solution and stir magnetically at room temperature for 30 min. A mixture was obtained, and the mixture was mixed with the polyaniline fiber membrane fragments in a weight ratio of 25:1 and then incubated for 2 hours. Mix and shake, then take out the polyaniline fiber membrane fragments and place them in an oven for 3 h incubation. The polyaniline fiber membrane fragments were then incubated with the mixture in a 15:1 ratio. The mixture was mixed in the ratio of 0.1 to 0.25, heated in a water bath for 4 hours, and the polyaniline fiber membrane fragments were taken out and dried to confirm the activity. The activated polyaniline fiber membrane fragments were obtained and then treated with 5:1 weight ratio. The debris and the laccase solution were mixed and stirred to a mass concentration of 20 mg / mL. Polyaniline-BiVO4 / ZnO fiber membrane metallokernel laccase obtained from polyaniline fiber The size of the membrane fragments was 5 mm, the average thickness of the polyaniline fiber membrane fragments was 2 mm, and the mixture The shaking rotation speed of the shaker was 1000 rpm, and the incubation temperature in the oven was 40 The temperature of the water bath was 100°C, and the polyaniline fiber membrane fragments were wrapped around the activated polyaniline fiber membrane. The rotation speed of the gelatin solution was 400 rpm. Polyaniline-BiVO4 / ZnO fibrous membrane metallokernel laccase and boundary during step S3 The application sequence of surfactant-based laccase is as follows: first, surfactant-based After laccase was applied to the soil by layer infiltration, polyaniline-BiVO4 / Zn O Fiber membrane metallokernel laccase was applied to the soil as a layer, and surfactant-based The permeation rate of laccase in the water was 0.6 L / h, and the permeation volume was 3 L / m 2 Each interface activity The spacing between the permeation layers of each surfactant-based laccase was 15 cm. The permeation layer of laccase and the polyaniline-BiVO4 / ZnO fibrous membranes containing metal kernel laccase The laying layers correspond one-to-one. Example 12 In this example, polyaniline-BiVO4 / was applied using the application device in step S3 of Example 1. The ZnO fiber membrane metallokernel laccase and surfactant-based laccase were applied, as shown in Figure 1. As shown, the application device includes a cartridge 1 disposed in a hollow space. To place metallokernel laccase inside the polyaniline-BiVO4 / ZnO fibrous membrane, A plurality of dispenser assemblies are provided for dispensing the ink from the inkjet printer, each of the dispenser assemblies being a cartridge. distributed in a spiral shape along the axial direction of the cartridge 1, As shown in Figures 2-4, the dispenser assembly consists of an outer slider (2), and a front The slider includes an inner slider (3) located inside the outer slider (2) and slidably connected thereto. The front end of the outer slider (2) is inserted into the first slider provided on the side wall of the cartridge (1). The rear end of the outer slider (2) is slidably connected to the cartridge (1). ) on the inner wall of the outer slider (2), and fixed blocks ( The front end of the fixed block (21) is supported by a first spring (22) on the cartridge. The front end of the fixed block (21) is connected to the inner wall of the ridge (1), and the rear end of the fixed block (21) is connected to the telescopic rod (23 ) and is connected to the rear end inner wall of the cartridge (1) through the front of the inner slider (3). The end is slidably inserted into a second slot (24) provided on the front wall of the outer slider (2). The rear end of the inner slider (3) is connected to the outer slider (2) through a cavity (25) and abutting against the rear end of the cavity (25), A third slot (26) is provided, and a porous baffle (27) is provided at the end of the third slot (26). The rear end of the inner slider (3) is connected to the porous backing (31) via a second spring (31). Connected to the center of the full (27), As shown in Figs. 2, 3 and 5, the inner slider 3 was coated with polyaniline-BiVO4 / ZnO fiber membrane metal kernel laccase placement groove 32 is provided, and the lower A permeable membrane is provided on the surface of the arrangement groove 32, and a magnetic attraction disk 33 is provided between the two permeable membranes. The inner slider 3 has an engagement block 34 provided on both the upper and lower sides of the rear end thereof. The lock 34 is slidably connected to the slide grooves 28 provided on both the upper and lower sides of the inner cavity of the outer slider 2. An air inlet 13 and a liquid inlet 14 are provided at the top of the cartridge 1. When the drill bit 15 is provided at the bottom of the edge 1 and the telescopic rod 23 is compressed to its shortest position, The front end of the outer slider (2) fits into the first slot (11), and the second spring (31) When the second slot (24) is fully extended, the positioning groove (32) is fully extended and the second slot (24) is fully extended. ) outside the As shown in FIGS. 1 and 2, the cartridge corresponding to the tip of each of the dispenser assemblies The inner wall of the ridge (1) is permeated with a surfactant-based laccase to infiltrate the soil. A hole (12) is provided in each of the holes, and a third spring (41) is provided on both the upper and lower sides of the penetration hole (12). The two third springs (41) are provided at their tips with a restricting spring for blocking the penetration hole (12). The limit baffle (4) is connected together, and when the third spring (41) naturally expands, the limit baffle The flues (4) are fitted to the rear end of the inner slider (3), and the total infiltration amount of all the infiltration holes 12 is The flow rate is 0.5L / h. Example 13 This example differs from Example 12 in the following respects: The number of dispenser assemblies is 6. This indicates that the number of outer sliders 2 and permeation holes 12 is six, and the present invention is based on the claims. In 4, the spacing between each of the six surfactant-based laccase permeation layers is 10 to 15 cm. Each (6) surfactant-based laccase permeation layer is each (6) polyaniline- The BiVO4 / ZnO fiber membrane corresponds to the laying layer of metal kernel laccase. Principle of Operation: The principle of operation of the application device of the present invention in conjunction with the method of the present invention is further described below. When using the cartridge, first insert the cartridge 1 through the drill bit 15 into the polycyclic aromatic hydrocarbon contaminated soil. Then, a surfactant-based laccase is injected through the liquid inlet 14. The soil is treated by infiltrating the soil through the infiltration hole 12 until the infiltration amount reaches the specified amount in step S3. After reaching the limit, pressurizing air is injected from the air inlet 13, and the restricting baffle 4 is pressed under the action of the air pressure. 3, the third spring 41 is compressed to close the penetration hole 12, and at the same time, the outer slider 2 is inserted into the first slot 11, compressing the first spring 22 and pulling the telescopic rod 23, When the inner slider 3 reaches its maximum extension, the inner slider 3 slides outward along the second slot 24. , the second spring 31 is pulled, and the arrangement groove 32 extends to the outside of the outer slider 2, and the arrangement groove 32 Polyaniline-BiVO4 / ZnO fibrous membranes containing metallokernel laccase in soil Treating polycyclic aromatic hydrocarbons in soil by contact. In addition, the newly prepared polyaniline-BiVO4 / ZnO was placed above the placement groove 32. The fiber membrane metal kernel laccase is arranged, and the polyaniline- After the BiVO4 / ZnO fiber membrane metallokernel laccase was placed in the cartridge, the soil was purified. The gas inside the slider 1 is exhausted, and the outer slider 2 and the inner slider 3 are moved by the action of the two springs. 3 was recovered, and the cartridge 1 was taken out and polyaniline-BiVO4 / ZnO fiber membrane gold was Extraction and recovery of laccase from polyaniline-BiVO4 / ZnO fibrous membrane metallization The method for recovering kernel laccase is as follows: The polyaniline-BiVO 2 and the inner slider 3 are pushed out, and the upper and lower sides of the arrangement groove 32 are 4 / ZnO fiber membrane metallokernel laccase was collected, and polyaniline-B was placed above the collection groove 32. The iVO4 / ZnO fiber membrane metallokernel laccase was treated and reused, and the bottom of the collection groove 32 The laccase is treated again depending on the number of times it is reused, and then, The required polyaniline-BiVO4 / ZnO fibrous membrane metal kernel laccase was placed again. Use it for soil purification next time or in the next location. Experimental Example 1 The methods in Examples 1 to 3 were applied to the detection of polycyclic aromatic hydrocarbons in actual soil, and the surfactant-based Benzo(a)anthracene in soils as a function of mass concentration of laccase in laccase solution. The effect of the fluoride on the degree of decomposition was observed, and the results are shown in Table 1. Table 1. Decomposition rate of benzo(a)anthracene in soil by the methods in Examples 1 to 3

[0009] TIFF2025017289000002.tif47145

[0010] As can be seen from the data in Table 1, the mass concentration of the laccase solution was set to 100-150 mg / L. The effect on the decomposition rate of benzo(a)anthracene in soil was large, and laccase When the mass concentration of the solution is 150 to 200 mg / L, the effect is small and the cost is high. Therefore, the mass concentration of the laccase solution in the surfactant-based laccase in Example 1 is preferred. stomach. Experimental Example 2 The methods in Examples 1, 4 to 8 were applied to the actual detection of polycyclic aromatic hydrocarbons in soil, and polyaniline was Phosphorus-BiVO4 / ZnO fibrous membrane metal kernel laccase and surfactant-based lacquer The effect of the ratio of fluoranthene to benzo(b)fluoranthene in soil was observed. The results are shown in Table 2. Table 2. Decomposition rate of benzo(b)fluoranthene in soil by the methods in Examples 1, 4 to 8

[0011] TIFF2025017289000003.tif68161

[0012] As can be seen from the data in Table 2, the polyaniline-BiVO4 / ZnO fibrous membrane metal kernel The ratio of laccase to surfactant-based laccase must be in the appropriate range. If the amount is too small, the decomposition rate may decrease. Since the decomposition rate is outside the bright range, the decrease in the decomposition rate is most significant compared to Example 1. Polyaniline-BiVO4 / ZnO fibrous membrane metallokernel laccase and surfactant-based A ratio of laccase to glycerol is preferred. Experimental Example 3 In addition, the formation of immobilized laccase by Fe3O4@SiO2-chitosan immobilized laccase can also provide some control effect on polycyclic aromatic hydrocarbons in water and soil. As a comparative example, in Comparative Example 1, Fe3O4@SiO2-chitosan immobilized lacquer The immobilized laccase was used to treat phenanthrene contaminants in real soil. In Comparative Example 2, the polyaniline-BiVO4 / ZnO fiber membrane metal kernel in Example 1 was used. Instead of laccase, the above Fe3O4@SiO2-chitosan immobilized laccase was used. The immobilized laccase was used to treat phenanthrene contaminants in real soil. The results are shown in Table 3. As shown in. Table 3. Benzo(a)anthracene in soil by the methods in Example 1, Comparative Examples 1 and 2 Decomposition rate

[0013] TIFF2025017289000004.tif50161

[0014] As can be seen from the data in Table 3, under the same soil sample and dosage conditions, the treatment method of the present invention The decomposition rate of phenanthrene in soil by the method is better than that of the two comparative examples, which is the reason why the present invention Polyaniline-BiVO4 / ZnO fibrous membranes containing metallokernel laccase and surfactant-based Rational combination of laccases effectively improves the degradation rate of phenanthrene contaminants in soil This shows that it is possible to effectively improve At the same time, as can be seen from the above three sets of experiments, the contamination of soil with various polycyclic aromatic hydrocarbons However, the method of the present invention can exert a good decomposition effect. Experimental Example 4 The methods in Examples 1 and 9 were applied to the actual detection of polycyclic aromatic hydrocarbons in soil, and the secondary regeneration Application of polyaniline-BiVO4 / ZnO fibrous membrane to metallokernel laccase and surfactant-based After resetting the ratio of laccase to cellulose, the removal effect of various pollutants in soil was observed and implemented. Compare with the method of Example 1 and also compare with Comparative Example 3, in which the method in Example 9 was used. The polyaniline-BiVO4 / ZnO fibrous membrane metallokernel laccase was then reused. The results are shown in Table 4. Table 4. Benzo(a)anthracene in soil by the methods in Examples 1, 9 and Comparative Example 3. Decomposition rate of benzo(b)fluoranthene

[0015] TIFF2025017289000005.tif51158

[0016] As can be seen from the data in Table 4, polyaniline-BiVO4 was synthesized using the method in Example 9. / ZnO fiber membrane metallokernel laccase recovery and comparison with surfactant-based laccase When the rate was adjusted again and reused, the removal rate of contaminants in the soil did not change significantly, which is It is shown that the method of Example 9 of the present invention can be carried out, and the apparatus of the present invention can be used in combination with the method of the present invention. In Comparative Example 3, the removal rate was significantly increased when the material was reused. Therefore, one reuse according to the method in Example 9 is sufficient, and multiple reuse is not recommended. Not recommended.

Claims

1. Step S1: Polyaniline-BiVO ４ Preparation of ZnO fiber membrane metallokernel laccase :Weight part, BiVO ４ Solid powder 1-2 parts, (CH ３ COO) ２ Zn solid powder 5-6 parts, Add 4 parts of NaOH solid powder to 15 to 20 parts of 75% ethanol solution at room temperature. The mixture was stirred magnetically for 30 minutes to obtain a mixture, and the mixture and the polyaniline fiber membrane fragments were mixed together for 15 to 20 minutes. The mixture was mixed in a weight ratio of 5:1 and shaken for 1 to 2 hours. After that, the polyaniline fiber membrane fragments were taken out. Then, place in an oven and incubate for 2 to 3 hours. Then, mix with the mixture and incubate. The polyaniline fiber membrane fragments were mixed in a weight ratio of 10-15:1, heated in a water bath for 3-4 hours, and The polyaniline fiber membrane fragments were taken out and dried to obtain activated polyaniline fiber membrane fragments. The activated polyaniline fiber membrane fragments and the laccase solution were mixed and stirred in a weight ratio of 5:1; The mass concentration of the laccase solution was 5 to 20 mg / mL, and polyaniline-BiVO ４ / Zn Obtaining an O-fiber membrane metallokernel laccase; Step S2, Preparation of surfactant-based laccase: rhamnolipid, laccase solution and deionized water to obtain surfactant-based laccase, and the obtained surfactant-based In the laccase of the above-mentioned, the mass concentration of rhamnolipid is 300 to 500 mg / L, A step of adjusting the mass concentration of the laccase solution to 100 to 200 mg / L; Step S3: Polyaniline-BiVO in soil ４ / ZnO fiber membrane metal kernel lacquer Application of laccase and surfactant-based laccase: Metal kernels in soil at a weight ratio of 3:5-10. Laccase and surfactant-based laccase are applied at a depth of 20-80 cm. and a step of: A method for removing polycyclic aromatic hydrocarbons from soil.

2. The method further comprises a step S4 of recovering and cycling the metallokernel laccase, Step S4, Metal kernel laccase recovery cycle: Step S3 through application device Polyaniline-BiVO applied at ４ / Recovering the ZnO fiber membrane metal kernel laccase; Recovered polyaniline-BiVO ４ / ZnO fiber membrane metal kernel laccase in deionized water After washing with 1000 ml of water, the recovered polyaniline-BiVO4 can be applied to the soil in step S3 again. ４ / ZnO fiber membrane metallokernel laccase and surfactant-based laccase in a weight ratio of 4:7 2. The method according to claim 1, wherein the composition is applied to the soil in a concentration of 0.1 to 100% by weight.

3. The size of the polyaniline fiber membrane fragments in step S1 is 0.5 mm to 5 mm. The thickness of the polyaniline fiber membrane fragments is 0.5 μm to 2 mm, and the shaking rotation speed of the mixing and shaking is The rotational speed was 1000 rpm, the incubation temperature in the oven was 400° C., and the water bath heating The temperature of the activated polyaniline fiber membrane was 100° C., and the activated polyaniline fiber membrane fragments and the laccase solution were mixed and stirred.

2. The method according to claim 1, wherein the stirring speed is 400 rpm.

4. BiVO in step S1 ４ The method for preparing the solid powder is as follows: 1 mol of Bi(NO ３ ) ３ ・5H ２ Take O and dissolve it in 20 mL of ethylene glycol solution. The mixture was dissolved and stirred magnetically for 60 min to obtain mixed solution A, and 0.01 mol of NH ４ V.O. ３ Take The mixture was dissolved in 20 mL of deionized water and stirred magnetically for 60 min to obtain mixture B. The mixture was added dropwise to the mixed solution A and magnetically stirred for 60 minutes, then placed in a reactor and reacted at 150°C for 10 hours. After cooling to room temperature in the furnace, the product was placed in a centrifuge and centrifuged to separate BiVO. ４ Obtaining a solid 2. The method according to claim 1, characterized in that the product is prepared by vacuum drying at 75°C for 2 hours.

5. Polyaniline-BiVO in step S3 ４ / ZnO fiber membrane metal kernel lacquer The application sequence of laccase and surfactant-based laccase is as follows: first, surfactant After applying the laccase-based solution to the soil by layer infiltration, the polyaniline-BiVO ４ / ZnO fiber membrane metal kernel laccase is applied inside the soil as a layer laying, and the interface activity The permeation rate of the laccase-based agent is 0.3-0.6 L / h, and the permeation volume is 2-3 L / h. m ２ and the spacing between each surfactant-based laccase impregnated layer is 10-15 cm. , each surfactant-based laccase-permeated layer and each polyaniline-BiVO ４ / ZnO fiber The method according to claim 1, characterized in that the layers of the membrane metallokernel laccase correspond one-to-one. Method of posting.

6. In step S3, polyaniline-BiVO is applied using an application device. ４ / ZnO fiber membrane metal The application device is hollow. The cartridge (1) includes a polyaniline-Bi V.O. ４ / Multiple dispenser applicators for disposing ZnO fiber membrane metal kernel laccase Each of the dispenser assemblies is arranged in the axial direction of the cartridge (1). Distributed in a spiral along the The dispenser assembly includes an outer slider (2), and a The inner slider (3) is located inside and slidably connected to the outer slider (2 The front end of the cartridge (1) is slidably inserted into a first slot (11) provided in the side wall of the cartridge (1). The rear end of the outer slider (2) extends to the inner wall of the cartridge (1), and the outer A fixing block (21) is provided on each of the upper and lower sides of the rear end of the side slider (2), The front end of the fixed block (21) is connected to the inside of the front end of the cartridge (1) via a first spring (22). The rear end of the fixed block (21) is connected to the cartridge through a telescopic rod (23). The rear end of the inner slider (3) is connected to the inner wall of the outer slider ( 2) is slidably connected to a second slot (24) provided on the front wall of the inner slider The rear end of the outer slider (3) extends into a cavity (25) provided inside the outer slider (2). The cavity (25) is abutted against a rear end thereof, and a third slot (26) is provided at the rear end of the cavity (25). A porous baffle (27) is provided at the tip of the third slot (26), and the inner slider The rear end of the porous baffle (27) is connected to the center of the porous baffle (27) via a second spring (31). R, On the inner wall of the cartridge (1) corresponding to the tip of each of the dispenser assemblies, Each of the soils had an infiltration hole (12) for infiltrating the surfactant-based laccase into the soil. It was kicked, The upper and lower sides of the inner slider (3) are coated with polyaniline-BiVO ４ / ZnO fiber membrane gold A placement groove (32) for placing the genus Kernel laccase is provided, and the lower placement groove ( A permeable membrane is provided on the surface of the magnetic disk (32), and a magnetic attraction disk (33) is provided between the two permeable membranes. The inner slider (3) has engagement blocks (34) on both the upper and lower sides of its rear end. The engagement blocks (34) are provided on both the upper and lower sides of the inner cavity of the outer slider (2). The cartridge (1) is slidably connected to the sliding groove (28) and has an air inlet (1) at the top of the cartridge (1). 3) and a liquid inlet (14), and a drill bit (15) at the bottom of the cartridge (1).

3. The method of claim 2, further comprising:

7. When the telescopic rod (23) is compressed to the shortest position, the front end of the outer slider (2) When the second spring (31) is fully extended by fitting into the first slot (11), the positioning groove (32) is fully extended and located outside of said second slot (24); A third spring (41) is provided on both the upper and lower sides of the penetration hole (12), and the two third springs (41) are A restricting baffle (4) for blocking the infiltration hole (12) is jointly connected to the tip of the spring (41). When the third spring (41) is naturally expanded, the limiting baffle (4) moves toward the inner slide. The number of dispenser assemblies is 5 to 6, and each dispenser is fitted to the rear end of the slider (3). a total permeation rate of all the permeation holes (12) is 0.3 to 0.6 L / h.

6. The method according to claim 6.

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