A laccase composite material, a preparation method and application thereof, and a treatment method for phenol-containing wastewater

Abstract

The present application belongs to the technical field of wastewater treatment, and provides a laccase composite material, a preparation method and application thereof, and a treatment method for phenol-containing wastewater. The laccase composite material provided by the present application comprises a substrate and laccase attached to the substrate; the substrate is foamed carbon, the specific surface area of the foamed carbon is 150-200 m 2 / g; the minimum size of the foamed carbon is greater than or equal to 0.2 cm; and the loading amount of the laccase is 1-10 wt%. The present application controls the minimum size of the substrate foamed carbon to be greater than or equal to 0.2 cm, so that the laccase composite material is easy to separate from water and easy to recycle; at the same time, the specific surface area of the foamed carbon is controlled to be 150-200 m 2 / g, and the loading amount of the laccase is 1-10 wt%, which also ensures the high degradation efficiency of the laccase composite material on phenolic compounds.

Description

Technical Field

[0001] This invention relates to the field of wastewater treatment technology, and in particular to a laccase composite material, its preparation method and application, and a method for treating phenol-containing wastewater. Background Technology

[0002] The presence of phenolic compounds poses a serious threat to the ecological environment and human health. Phenolic compounds are highly toxic and stable; even small amounts entering the environment can cause significant public health risks, and they are difficult to remove from the environment through self-cleaning. Traditional physicochemical methods, such as adsorption and chemical oxidation, while effective in some cases, are often accompanied by high energy consumption, high costs, and the potential for secondary pollution. Therefore, finding an efficient and environmentally friendly treatment method is of paramount importance.

[0003] Laccase is a copper-containing polyphenol oxidase widely found in various organisms. Due to its broad substrate spectrum and high catalytic efficiency, it is considered an ideal biocatalyst for treating phenolic compounds. However, in practical applications, free laccase faces problems such as poor stability, easy inactivation, and difficulty in recovery and reuse. These issues limit the widespread application of laccase in wastewater treatment. To address these problems, immobilization technology has been introduced, aiming to improve its stability and reusability, and enhance its catalytic efficiency by immobilizing laccase on a support.

[0004] Existing technologies immobilize laccase on a carrier, resulting in laccase composite materials that are difficult to separate from water. Summary of the Invention

[0005] In view of this, the purpose of this invention is to provide a laccase composite material, its preparation method and application, and a method for treating phenol-containing wastewater. The laccase composite material provided by this invention is easily separated from water and has high degradation efficiency for phenolic compounds.

[0006] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0007] This invention provides a laccase composite material, comprising a matrix and laccase attached to the matrix;

[0008] The matrix is ​​foamed carbon, and the specific surface area of ​​the foamed carbon is 150-200 m². 2 / g;

[0009] The minimum size of the foamed carbon is greater than or equal to 0.2 cm;

[0010] The loading amount of the laccase is 1–10 wt%.

[0011] Preferably, the method for preparing the foamed carbon includes the following steps:

[0012] A hydrothermal system is obtained by mixing lignin and an inorganic alkaline solution and then subjecting the mixture to hydrothermal treatment.

[0013] The hydrothermal system was subjected to acid precipitation to obtain a carbon precursor;

[0014] The carbon precursor is heat-treated to obtain the foamed carbon.

[0015] Preferably, the inorganic alkali in the inorganic alkali solution includes sodium hydroxide and / or potassium hydroxide, the concentration of the inorganic alkali solution is 0.8-1.2 mol / L, and the ratio of lignin to inorganic alkali solution is 1 g: 40-50 mL;

[0016] The hydrothermal treatment is performed at a temperature of 160–200°C for 5–12 hours.

[0017] Preferably, the reagent for acid precipitation is hydrochloric acid, and the concentration of hydrochloric acid in the acid precipitation system is 0.4 to 0.6 mol / L.

[0018] Preferably, the heat treatment includes performing a first heat treatment and a second heat treatment sequentially;

[0019] The temperature of the first heat treatment is 600-700℃, the holding time is 1-3h, and the heating rate to the temperature of the first heat treatment is 5-15℃ / min.

[0020] After the first heat treatment, the temperature is directly increased to carry out the second heat treatment;

[0021] The temperature of the second heat treatment is 750-800℃, the holding time is 2-4h, and the heating rate to the temperature of the second heat treatment is 5-15℃ / min.

[0022] This invention also provides a method for preparing the laccase composite material described in the above technical solution, comprising the following steps:

[0023] After being impregnated in a laccase solution, the foamed carbon is immobilized to obtain the laccase composite material.

[0024] Preferably, the concentration of the laccase solution is 0.5–1.5 g / L;

[0025] After the foamed carbon is impregnated in laccase solution, the concentration of foamed carbon in the resulting system is 1-2 g / L.

[0026] Preferably, the immobilization time is 0.5 to 3 hours, and the immobilization is carried out under oscillation conditions with an oscillation rate of 100 to 200 rpm.

[0027] The present invention also provides the application of the laccase composite material described in the above technical solution or the laccase composite material prepared by the above technical solution in the degradation of phenolic compounds.

[0028] This invention also provides a method for treating phenol-containing wastewater, comprising the following steps:

[0029] The laccase composite material described in the above technical solution or the laccase composite material prepared by the above technical solution is placed in wastewater containing phenolic compounds for treatment.

[0030] The concentration of phenolic compounds in the wastewater containing phenolic compounds is 50–100 mg / L;

[0031] The concentration of the laccase composite material in wastewater containing phenolic compounds is greater than or equal to 0.2 mg / mL.

[0032] This invention provides a laccase composite material.

[0033] This invention controls the minimum size of the matrix foam carbon to be greater than or equal to 0.2 cm, making the laccase composite material easy to separate from water and easy to recycle; at the same time, it controls the specific surface area of ​​the foam carbon to be 150-200 m². 2 The laccase loading is 1-10 wt%, which also ensures the high degradation efficiency of laccase composite materials for phenolic compounds.

[0034] The present invention also provides a method for preparing the laccase composite material described in the above technical solution. The preparation method provided by the present invention is simple to operate.

[0035] Furthermore, the present invention limits the concentration of the laccase solution to 0.5–1.5 g / L; after the foamed carbon is impregnated in the laccase solution, the concentration of the foamed carbon in the resulting system is 1–2 g / L, so that the laccase can be more uniformly loaded on the foamed carbon, thereby improving the degradation efficiency of phenolic compounds by the laccase composite material. Attached Figure Description

[0036] Figure 1 A scanning electron microscope image of the lignin-based foam carbon obtained in Example 1;

[0037] Figure 2 This is a photograph of the lignin-based foam carbon obtained in Example 1. Detailed Implementation

[0038] This invention provides a laccase composite material, comprising a matrix and laccase attached to the matrix;

[0039] The matrix is ​​foamed carbon, and the specific surface area of ​​the foamed carbon is 150-200 m². 2 / g;

[0040] The minimum size of the foamed carbon is greater than or equal to 0.2 cm;

[0041] The loading amount of the laccase is 1–10 wt%.

[0042] Unless otherwise specified, the raw materials used in this invention are preferably commercially available products.

[0043] The laccase composite material provided by this invention includes a matrix, wherein the matrix is ​​carbon foam, and the specific surface area of ​​the carbon foam is 150-200 m². 2 / g, preferably 150m 2 / g、155m 2 / g、160m 2 / g、170m 2 / g、180m 2 / g、190m 2 / g or 200m 2 / g. In this invention, the minimum size of the foamed carbon is greater than or equal to 0.2 cm, and more preferably, the minimum size of the foamed carbon is 0.2 to 10 cm.

[0044] In this invention, the method for preparing the foamed carbon preferably includes the following steps:

[0045] A hydrothermal system is obtained by mixing lignin and an inorganic alkaline solution and then subjecting the mixture to hydrothermal treatment.

[0046] The hydrothermal system was subjected to acid precipitation to obtain a carbon precursor;

[0047] The carbon precursor is heat-treated to obtain the foamed carbon.

[0048] This invention involves mixing lignin and an inorganic alkali solution, followed by hydrothermal treatment to obtain a hydrothermal system. In this invention, the inorganic alkali in the inorganic alkali solution preferably includes sodium hydroxide and / or potassium hydroxide, more preferably sodium hydroxide. In this invention, the concentration of the inorganic alkali solution is preferably 0.8–1.2 mol / L, specifically preferably 0.8 mol / L, 0.9 mol / L, 1 mol / L, 1.1 mol / L, or 1.2 mol / L. In this invention, the preferred ratio of lignin to inorganic alkali solution is 1 g:40–50 mL, specifically preferably 1 g:40 mL, 1 g:45 mL, or 1 g:50 mL.

[0049] In this invention, the mixing of lignin and inorganic alkaline solution is preferably carried out under stirring conditions. The stirring speed is preferably 100-150 rpm, specifically 100 rpm, 110 rpm, 120 rpm, 130 rpm, 140 rpm or 150 rpm; the temperature is preferably 20-30℃, specifically 20℃, 25℃ or 30℃; and the time is preferably 20-40 min, specifically 20 min, 30 min or 40 min.

[0050] In this invention, the hydrothermal treatment temperature is preferably 160–200°C, specifically 160°C, 170°C, 180°C, 190°C, or 200°C; the time is preferably 5–12 hours, specifically 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, or 12 hours. In this invention, the hydrothermal treatment can dissolve lignin and simultaneously remove insoluble impurities from the lignin, thus purifying it.

[0051] In this invention, the hydrothermal treatment is preferably carried out in a polytetrafluoroethylene digestion tank.

[0052] After the hydrothermal treatment, the present invention preferably further includes: cooling the obtained hydrothermal treatment solution to room temperature to obtain the hydrothermal system.

[0053] After obtaining the hydrothermal system, the present invention performs acid precipitation on the hydrothermal system to obtain a carbon precursor.

[0054] In this invention, the reagent for acid precipitation is preferably hydrochloric acid, and the concentration of hydrochloric acid in the acid precipitation system is preferably 0.4 to 0.6 mol / L, specifically preferably 0.4 mol / L, 0.5 mol / L or 0.6 mol / L.

[0055] In this invention, the acid precipitation temperature is preferably 20–30°C, specifically 20°C, 25°C, or 30°C; the time is preferably 20–40 min, specifically 20 min, 30 min, or 40 min. In this invention, the acid precipitation is preferably carried out under stirring conditions, and the stirring speed is preferably 100–150 rpm, specifically 100 rpm, 110 rpm, 120 rpm, 130 rpm, 140 rpm, or 150 rpm. In this invention, the acid precipitation can precipitate dissolved lignin.

[0056] Following acid precipitation, the present invention preferably further includes: centrifuging the obtained acid precipitation solution to collect the solid; and sequentially washing and drying the solid to obtain the carbon precursor. In this invention, the centrifugation speed is preferably 2000–40000 rpm, specifically preferably 2000 rpm, 3000 rpm, or 4000 rpm. In this invention, the washing reagent is preferably water, more preferably deionized water; the washing is preferably performed three times, and the washing is effective in removing acid and salt. In this invention, the drying temperature is preferably 80°C, and the drying time is preferably 12 hours.

[0057] After obtaining the carbon precursor, the present invention performs heat treatment on the carbon precursor to obtain the foamed carbon.

[0058] In this invention, the heat treatment preferably includes performing a first heat treatment and a second heat treatment in sequence.

[0059] In this invention, the temperature of the first heat treatment is preferably 600–700°C, specifically preferably 600°C, 610°C, 620°C, 630°C, 640°C, 650°C, 660°C, 670°C, 680°C, 690°C, or 700°C; the holding time is preferably 1–3 hours, specifically preferably 1 hour, 2 hours, or 3 hours; the heating rate to the temperature of the first heat treatment is preferably 5–15°C / min, specifically preferably 5°C / min, 10°C / min, or 15°C / min. In this invention, the first heat treatment is preferably carried out under a protective atmosphere, preferably nitrogen. In this invention, the first heat treatment is preferably carried out in a tube furnace.

[0060] After the first heat treatment, the present invention preferably performs a second heat treatment by directly raising the temperature.

[0061] In this invention, the temperature of the second heat treatment is preferably 750–800°C, specifically preferably 750°C, 760°C, 770°C, 780°C, 790°C, or 800°C; the holding time is preferably 2–4 hours, specifically preferably 2 hours, 3 hours, or 4 hours; the heating rate to the temperature of the second heat treatment is preferably 5–15°C / min, specifically preferably 5°C / min, 10°C / min, or 15°C / min. In this invention, the second heat treatment is preferably carried out under a protective atmosphere, preferably nitrogen. In this invention, the second heat treatment is preferably carried out in a tube furnace.

[0062] After the heat treatment, the present invention preferably cools to room temperature to obtain the foamed carbon.

[0063] The laccase composite material provided by the present invention comprises laccase attached to the matrix. In the present invention, the loading amount of the laccase is 1-10 wt%, specifically preferably 1 wt%, 2 wt%, 3 wt%, 4 wt%, 4.2 wt%, 5 wt%, 6 wt%, 7 wt%, 8 wt%, 8.1 wt%, 9 wt%, 9.8 wt%, or 10 wt%.

[0064] In one specific embodiment of the present invention, the laccase gene is preferably AA1_Bin30_Lac. In this invention, the laccase gene is preferably selected from metagenomics (BioProject ID: PRJNA785376) as a laccase gene with strong structural stability. In this invention, the method for obtaining the laccase gene preferably includes gene synthesis, heterologous expression, and isolation and purification to obtain recombinant laccase. In one specific embodiment of the present invention, the laccase gene is preferably obtained according to patent publication number CN114574453A.

[0065] This invention also provides a method for preparing the laccase composite material described in the above technical solution, comprising the following steps:

[0066] After being impregnated in a laccase solution, the foamed carbon is immobilized to obtain the laccase composite material.

[0067] In this invention, the concentration of the laccase solution is preferably 0.5 to 1.5 g / L, and more preferably 0.5 g / L, 1 g / L or 1.5 g / L.

[0068] In this invention, after the foamed carbon is impregnated in laccase solution, the concentration of the foamed carbon in the resulting system is preferably 1 to 2 g / L, specifically preferably 1 g / L, 1.5 g / L or 2 g / L.

[0069] In this invention, the immobilization time is preferably 0.5 to 3 hours, specifically preferably 0.5 hours, 1 hour, 1.5 hours, 2 hours, 2.5 hours or 3 hours; the immobilization is preferably carried out under oscillation conditions, and the oscillation rate is preferably 100 to 200 rpm, specifically preferably 100 rpm, 150 rpm or 200 rpm.

[0070] After immobilization, the present invention preferably further includes: washing, separating and vacuum drying the obtained immobilized liquid in sequence to obtain the laccase composite material.

[0071] The present invention also provides the application of the laccase composite material described in the above technical solution or the laccase composite material prepared by the above technical solution in the degradation of phenolic compounds.

[0072] This invention also provides a method for treating phenol-containing wastewater, comprising the following steps:

[0073] The laccase composite material described in the above technical solution or the laccase composite material prepared by the above technical solution is placed in wastewater containing phenolic compounds for treatment.

[0074] In this invention, the phenolic compounds are preferably one or more selected from hydroquinone, phenol, cresol, 2,4-dichlorophenol, 2,4,6-trichlorophenol and p-nitrophenol.

[0075] In this invention, the concentration of phenolic compounds in the phenolic wastewater is preferably 50-100 mg / L, and more preferably 50 mg / L, 60 mg / L, 70 mg / L, 80 mg / L, 90 mg / L or 100 mg / L.

[0076] In this invention, the pH value of the phenolic compound-containing wastewater is preferably 4.5 to 5.5, and more preferably 4.5, 5 or 5.5.

[0077] In this invention, the concentration of the laccase composite material in wastewater containing phenolic compounds is greater than or equal to 0.2 mg / mL, and more preferably 0.2 to 10 mg / mL.

[0078] In this invention, the processing is preferably carried out under oscillation conditions, and the oscillation speed is preferably 100 to 200 rpm, specifically 100 rpm, 150 rpm or 200 rpm.

[0079] After the treatment, the present invention preferably further includes: separating the obtained treatment liquid and collecting the laccase composite material.

[0080] In this invention, the separation method is preferably filtration.

[0081] In this invention, the laccase composite material is separated from the water body, which can reduce the secondary pollution of the water body by the laccase composite material.

[0082] The following detailed descriptions, in conjunction with embodiments, illustrate the laccase composite material, its preparation method, its application, and the treatment method for phenol-containing wastewater provided by this invention. However, these descriptions should not be construed as limiting the scope of protection of this invention.

[0083] Example 1

[0084] Preparation method of lignin-based foamed carbon:

[0085] (1) Dissolve 1g of lignin in 50mL of 1mol / L NaOH solution, stir at 25℃ for 30min, then transfer to a polytetrafluoroethylene digestion vessel and hydrothermally treat at 180℃ for 6h. Remove the polytetrafluoroethylene digestion vessel and cool to room temperature to obtain a black mixture, which is the hydrothermal system. Transfer the black mixture to a 500mL wide-mouth bottle for acid precipitation. Continuously add 50mL of 2mol / L hydrochloric acid to the black mixture to achieve a hydrochloric acid concentration of 0.5mol / L. Stir at 25℃ and 100rpm for 30min. After a brown precipitate appears, centrifuge at 3000rpm to collect the solid. Wash the brown precipitate three times with deionized water to remove hydrochloric acid and NaCl, then dry at 80℃ for 12h to obtain the carbon precursor.

[0086] (2) Take 2.0g of the above carbon precursor, place it in a tube furnace, and heat it to 600℃ for 2h at a heating rate of 10℃ / min under N2 atmosphere; continue heating to 800℃ at a heating rate of 10℃ / min and hold for 3h, then cool to room temperature to obtain lignin-based foam carbon.

[0087] The microstructure of the obtained lignin-based foam carbon was observed using a scanning electron microscope, and the results are as follows: Figure 1 As shown, from Figure 1 It can be seen that there are connecting channels between the pores, forming a connected pore network. The pores can be circular, elliptical, etc. The surface is rough with cracks, and the pores are evenly distributed on the material surface, forming a relatively regular structure.

[0088] Figure 2 A photograph of the resulting lignin-based foam carbon.

[0089] The specific surface area and pore size of lignin-based foamed carbon were determined using the BET method. The results showed that the specific surface area of ​​the obtained lignin-based foamed carbon was 155.5504 m². 2 / g, with an average pore size of 3.3μm.

[0090] Example 2

[0091] Preparation of laccase:

[0092] Acquisition of the laccase gene: A structurally stable laccase gene (AA1_Bin30_Lac) was screened from the metagenomics database (BioProject ID: PRJNA785376). Recombinant laccase was obtained by gene synthesis, heterologous expression, and isolation and purification according to the patent published in CN114574453A. The concentration was detected by SDS-PAGE and the result was 1.0 mg / mL.

[0093] Laccase activity and stability assay: The standard substrate ABTS was selected to determine the activity of laccase. The specific procedure was as follows: a 700 μL reaction system was set up, containing 600 μL Tris-HCl solution, 50 μL ABTS, and 50 μL of enzyme solution with a concentration of 1.0 mg / mL, and the pH of the reaction system was adjusted to 5.0; the reaction was carried out at 20℃, 25℃, and 30℃ for 5 min, and the change in absorbance was measured at 420 nm. The amount of enzyme required to oxidize 1 μmol / LABTS per minute was defined as one unit of laccase activity.

[0094] A 700 μL reaction system was set up, containing 600 μL Tris-HCl solution, 50 μL Labiscuits, and 50 μL of enzyme solution with a concentration of 1.0 mg / mL. The pH of the reaction system was adjusted to 4.5, 5.0, 5.5, and 6.0. The reaction was carried out at 25 °C for 5 min, and the change in absorbance was measured at 420 nm. The amount of enzyme required to oxidize 1 μmol / LABTS per minute was defined as one unit of laccase activity.

[0095] The results are shown in Table 1.

[0096] Table 1 Results of Laccase Activity Test

[0097]

[0098] As can be seen from Table 1, the enzyme activity is optimal when the temperature is between 25 and 30°C and the pH value is around 5.0.

[0099] Example 3

[0100] The lignin-based foam carbon obtained in Example 1 was cut into 1.0cm×1.0cm×0.2cm pieces and added to a laccase solution with an initial concentration of 0.5g / L at concentrations of 1.0g / L, 1.5g / L, and 2.0g / L. The laccase was immobilized by shaking in a shaker (150rpm) for 2 hours. After washing, separation, and vacuum drying, the laccase composite material was obtained.

[0101] The concentration of laccase in the laccase solution before and after the reaction was tested using the BCA method. The amount of enzyme adsorbed per unit carrier of each laccase composite material was calculated under different lignin-based foam carbon concentrations. The results showed that the laccase loading was high at a lignin-based foam carbon concentration of 2.0 g / L, which was 4.2 wt%.

[0102] The lignin-based foam carbon obtained in Example 1 was cut into 1.0cm×1.0cm×0.2cm pieces and added to laccase solutions with initial concentrations of 0.5g / L, 1.0g / L, and 1.5g / L at a concentration of 2.0g / L. The laccase was immobilized by shaking in a shaker (150rpm) for 2 hours. After washing, separation, and vacuum drying, the laccase composite material was obtained.

[0103] The concentration of laccase in the laccase solution before and after the reaction was tested using the BCA method. The amount of enzyme adsorbed per unit carrier of each laccase composite material was calculated at different initial laccase concentrations. The results showed that the laccase solution with an initial concentration of 1.5 g / L had a high laccase loading of 8.1 wt%.

[0104] The lignin-based foam carbon obtained in Example 1 was cut into 1.0cm×1.0cm×0.2cm pieces and added to a laccase solution with an initial concentration of 1.5g / L at a concentration of 2.0g / L. The solution was shaken in a shaker (150rpm) for 0.5h, 1h, 1.5h, 2.0h, and 3.0h respectively to complete the immobilization of laccase. After washing, separation, and vacuum drying, the laccase composite material was obtained.

[0105] The concentration of enzyme before and after the reaction was tested using the BCA method. The amount of enzyme adsorbed per unit carrier of each laccase composite material was calculated at different immobilization times. The results showed that the laccase loading was high at an immobilization time of 2 h, which was 9.8 wt%.

[0106] Example 4

[0107] The lignin-based foam carbon obtained in Example 1 was cut into 1.0cm×1.0cm×0.2cm pieces and added to a laccase solution with an initial concentration of 1.5g / L at a concentration of 2.0g / L. The mixture was shaken in a shaker (150rpm) for 2 hours to complete the immobilization of the laccase. After washing, separation, and vacuum drying, a laccase composite material was obtained, in which the laccase loading was 9.8wt%.

[0108] Application Example 1

[0109] Hydroquinone, a phenolic compound, was added to deionized water to form simulated phenol-containing wastewater with a phenolic compound concentration of 100 mg / L, and the pH value of the simulated phenol-containing wastewater was adjusted to 5.0.

[0110] The laccase composite material obtained in Example 4 was added to simulated phenol-containing wastewater at a concentration of 0.3 mg / mL and shaken at 150 rpm for 10 h. The concentration of phenolic compounds in the resulting simulated phenol-containing wastewater was 0.39 mg / L, which is lower than the national wastewater discharge standard (CJ343-2010) of 0.5–1.0 mg / L.

[0111] After the reaction is complete, the laccase composite material is directly removed from the simulated phenol-containing wastewater through physical separation and recycled for reuse.

[0112] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A laccase composite material, characterized in that, Includes the matrix and the laccase attached to the matrix; The matrix is ​​carbon foam, and the specific surface area of ​​the carbon foam is 150~200m². 2 / g; The minimum size of the foamed carbon is greater than or equal to 0.2 cm; The loading amount of laccase is 1~10 wt%; The method for preparing the foamed carbon includes the following steps: A hydrothermal system is obtained by mixing lignin and an inorganic alkaline solution and then subjecting the mixture to hydrothermal treatment. The hydrothermal system was subjected to acid precipitation to obtain a carbon precursor; The carbon precursor is heat-treated to obtain the foamed carbon; The hydrothermal treatment is performed at a temperature of 170~200℃ for 5~12 hours. The heat treatment includes performing a first heat treatment and a second heat treatment in sequence; The temperature of the first heat treatment is 600~700℃, the holding time is 1~3h, and the heating rate to the temperature of the first heat treatment is 5~15℃ / min. After the first heat treatment, the temperature is directly increased to carry out the second heat treatment; The temperature of the second heat treatment is 750~800℃, the holding time is 2~4h, and the heating rate to the temperature of the second heat treatment is 5~15℃ / min; The preparation method of the laccase composite material includes the following steps: After impregnation in laccase solution, foamed carbon is immobilized to obtain the laccase composite material. The concentration of the laccase solution is 1.5 g / L; After the foamed carbon is impregnated in laccase solution, the concentration of foamed carbon in the resulting system is 2 g / L. The immobilization time is 2 hours.

2. The laccase composite material according to claim 1, characterized in that, The inorganic alkali in the inorganic alkali solution includes sodium hydroxide and / or potassium hydroxide, the concentration of the inorganic alkali solution is 0.8~1.2 mol / L, and the ratio of lignin to inorganic alkali solution is 1g:40~50mL.

3. The laccase composite material according to claim 1, characterized in that, The reagent used for acid precipitation is hydrochloric acid, and the concentration of hydrochloric acid in the acid precipitation system is 0.4~0.6 mol / L.

4. The method for preparing the laccase composite material according to any one of claims 1 to 3, characterized in that, Includes the following steps: After impregnation in laccase solution, foamed carbon is immobilized to obtain the laccase composite material. The concentration of the laccase solution is 1.5 g / L; After the foamed carbon is impregnated in laccase solution, the concentration of foamed carbon in the resulting system is 2 g / L. The immobilization time is 2 hours.

5. The preparation method according to claim 4, characterized in that, The immobilization is performed under oscillation conditions, with the oscillation rate being 100~200 rpm.

6. A method for treating phenol-containing wastewater, characterized in that, Includes the following steps: The laccase composite material according to any one of claims 1 to 3 or the laccase composite material prepared by the preparation method according to any one of claims 4 to 5 is placed in wastewater containing phenolic compounds for treatment. The concentration of phenolic compounds in the wastewater containing phenolic compounds is 50~100 mg / L; The concentration of the laccase composite material in wastewater containing phenolic compounds is greater than or equal to 0.2 mg / mL.

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