Immobilization Of Laccase Research Articles

Laccase encapsulation immobilized in mesoporous ZIF-8 for enhancement bisphenol A degradation.

Endocrine disruptors (EDCs) such as bisphenol A (BPA) have many adverse effects on environment and human health. Laccase encapsulation immobilized in mesoporous ZIF-8 was prepared for efficient degradation of BPA. The ZIF-8 (PA) with highly ordered mesopores was synthesized using trimethylacetic acid (PA) as a template agent. On account of the improvement of skeletal stability by cross-linking agent glutaraldehyde, ZIF-8 (PA) realized laccase (FL) immobilization within the mesopores through encapsulation strategy. By replacing the template agent, the effect of pore size on the composite activity and immobilization efficiency by SEM characterization and kinetic analysis were investigated. Based on the physical protection of ZIF-8(PA) on laccase, as well as electrostatic interactions between substances and changes in surface functional groups (e.g. -OH, etc.), multifaceted enhancement including activity, stability, storability were engendered. FL@ZIF-8(PA) could maintain high activity in complex systems at pH 3-11, 10-70°C or in organic solvent containing system, which exhibited an obvious improvement compared to free laccase and other reported immobilized laccase. Combined with TGA, FT-IR and Zeta potential analysis, the intrinsic mechanism was elaborated in detail. On this basis, FL@ZIF-8(PA) achieved efficient removal of BPA even under adverse conditions (removal rates all above 55% and up to 90.28%), and was suitable for a wide range of initial BPA concentrations. Combined with the DFT calculations on the adsorption energy and differential charge, the mesoporous could not only improve the enrichment performance of BPA on ZIFs, but also enhance the interaction stability. Finally, FL@ZIF-8(PA) was successfully applied to the degradation of BPA in coal industry wastewater. This work provides a new and ultra-high performances material for the organic pollution treatment in wastewater.

Improvement of Laccase Production by Thielavia terrestris Co3Bag1. Enhancing the Bio-Catalytic Performance of the Native Thermophilic TtLacA via Immobilization in Copper Alginate Gel Beads.

A 32-fold increase in laccase activity production by the thermophilic biomass-degrading fungus T. terrestris Co3Bag1 was achieved when the microorganism was grown on a modified medium containing fructose, sodium nitrate, and copper. A 70 kDa laccase (TtLacA), produced under the above conditions, was purified, immobilized in copper alginate gel beads, and characterized. TtLacA, both free and immobilized enzymes, exhibited optimal activity at pH 3.0, at a temperature of 65 and 70 °C, respectively, although both displayed 70% of activity from 40 to 70 °C. Free and immobilized enzymes retained at least 80% of relative activity in the pH range from 3 to 4.6. Immobilized TtLacA manifested a 2.3-fold higher thermal stability than the free form of the enzyme at 60 and 70 °C. Immobilized TtLacA retained 95% initial activity for six consecutive reuse cycles at 60 °C, and also retained 86% of initial activity after 12 days of storage at 4 °C. Based on the biochemical features, thermophilic TtLacA may be an efficient enzyme for dye decolorization and other industrial applications at high temperatures or acidic conditions. This work represents the first report about the immobilization and biochemical characterization of a thermophilic laccase from a member of the genus Thielavia.

How can vanillin improve the performance of lignocellulosic biomass conversion in an immobilized laccase microreactor system?

Gentle and effective pretreatment is necessary to produce clean lignocellulosic biomass-based fuels. Herein, inspired by the efficient lignin degradation in the foregut of termites, the microreactor system using immobilized laccase and recoverable vanillin was proposed. Firstly, the co-deposition coating of dopamine, hydrogen peroxide and copper sulfate was constructed for laccase immobilization and a high immobilization efficiency of 87.0% was obtained in 30 min. After storage for 10 days, 82.2% activity was maintained in the laccase-loaded microreactor, which is 210.0% higher than free laccase. In addition, 6% (w/w) vanillin can improve lignin degradation in the laccase-loaded microreactor without impairing laccase activity, leading to a 47.3% increment in cellulose accessibility. Finally, a high cellulose conversion rate of 88.1% can be achieved in 1 h with glucose productivity of 2.62 g L−1 h−1. These demonstrated that the appropriate addition of vanillin can synergize with immobilized laccase to enhance the conversion of lignocellulosic biomass.

Functionalization of micropipette tips with hydrophobin-laccase chimera and application to the electrochemical determination of caffeic acid in tea samples

Performing on-site measurements is one of the main challenges of current (bio)chemical analysis. Micropipette tips, which are among the most common utilities in laboratories, can be useful and cheap elements for designing out-of-the-box innovative analytical biodevices. Furthermore, most of the analytical properties of these bioplatforms depend on the immobilization of the recognition element (e.g., enzymes). In this context, we have recently designed a chimeric protein in which laccase PoxA1b from the fungus Pleurotus ostreatus has been genetically fused to the class I hydrophobin Vmh2. The ability to self-assemble at hydrophobic-hydrophilic interfaces of hydrophobin, allows an easy laccase immobilization. In this work, the Vmh2-PoxA1b fusion protein was immobilized into common polypropylene micropipette tips to develop an innovative enzymatic bioplatform that can produce fast results in a simple way. As proof of applicability, the platform was applied to the determination of caffeic acid by enzymatic oxidation and further electrochemical reduction of the product on a screen-printed electrode, achieving a limit of detection of 1.4·10−6 M. It was then used for determining caffeic acid in tea samples.

Immobilization of Trametes trogii laccase on polyvinylpyrrolidone-coated magnetic nanoparticles for biocatalytic degradation of textile dyes

High cost and low operational stability are the most important challenges limiting the possible use of laccase in the removal of textile dyes. To overcome these challenges, in this study, polyvinylpyrrolidone (PVP)-coated magnetic nanoparticles (MNPs) were produced and characterized. To our knowledge, this is the first study to explore the feasibility of immobilizing a Trametes trogii laccase enzyme on Fe3O4/PVP MNPs. The characterization of samples and the successful immobilization of laccase were verified by characterization methods. Besides, the biochemical properties and stability of the Fe3O4/PVP/Lac were evaluated in terms of optimum pH, optimum temperature, thermostability, thermodynamic and kinetic parameters, storage stability, operational stability, and decolorization efficiency of two different textile dyes. The optimum activities were recorded at pH 2.5 °C and 30 °C. The Fe3O4/PVP/Lac displayed remarkable thermal stability and activation energy for denaturation, enthalpy, Gibbs free energy, and entropy results confirmed the enhanced stability of Fe3O4/PVP/Lac against high temperatures. Meanwhile, the Fe3O4/PVP/Lac retained about 58% of its original activity after seven consecutive reuses, while it retained up to 25% of its original activity after 28 d of storage at room temperature. Km and V max for the Fe3O4/PVP/Lac were calculated to be 126 µM and 211 µmol/min, respectively. Finally, after 8 and 6 cycles of repeated use, the Fe3O4/PVP/Lac still decolorized 32.34% and 32.23% of Remazol Brilliant Blue R (RBBR) and Indigo Carmine (IC), respectively. As envisioned, this study suggests a promising way to solve the problems of high price and poor operational stability of the enzyme during biocatalytic decolorization of textile dyes in wastewaters.

Preparation of Chitosan-Carbon Nanotubes Decorated with ZnO@laccase: Application in the Removal of Bisphenol A from Aqueous Solution

Bioremediation with immobilized enzymes has been established as the method for restoring contaminated ecosystems with the help of biological tools on a global scale. The purpose of this work is to immobilize the laccase enzyme for increasing the biocatalyst′s ability in order to remove bisphenol A (BPA). Initially, the nanocatalyst was prepared by immobilizing laccase on chitosan using carbon nanotubes decorated with zinc oxide. Subsequently, Fourier transformed infrared (FTIR) spectroscopy, X-ray diffraction (XRD), field emission scanning microscopy (FE-SEM), transmission electron microscopy (TEM), energy-dispersive X-ray spectroscopy (EDX), and dot mapping were employed to characterize the prepared nanocomposites. Laccase activity, immobilization yield (%), thermal and storage stability, and reusability of laccase immobilized on chitosan-coated carbon nanotubes (CNTs)/ZnO nanocomposites were also measured. The chitosan (CS)-CNTs/[email protected] nanocatalyst performance in different operational conditions (initial concentrations of bisphenol A, amount of nanobiocatalyst, pH, and irradiation time) on the photodegradation process of bisphenol A were also investigated and optimized using the response surface methodology (RSM)-central composite design (CCD) model. The RSM design and experiments demonstrated that 94.51% of BPA was removed under optimal conditions (initial BPA concentration of 100 mg L−1, nanocatalyst amount of 1 g L−1, pH=9, and 40 min of contact exposure). Laccase was a critical factor in this process, and after five recycling cycles, immobilized laccase retained 61% of its original activity, indicating that this nanobiocatalyst can be used in purification processes as a durable and robust catalyst.

Effects of co-modified biochar immobilized laccase on remediation and bacterial community of PAHs-contaminated soil.

Considering the stability and economy of immobilized enzymes, this study prepared co-modified biochar immobilized laccase product named Fe3O4@NaBC@GA@LC via orthogonal experimental design and explored its possibility of remediating polycyclic aromatic hydrocarbons (PAHs) contaminated soil in steel plants. Compared with the free laccase treatment, the relative activity of Fe3O4@NaBC@GA@LC remained 60% after 50 days of incubation at room temperature. The relative activity of Fe3O4@NaBC@GA@LC could still retain nearly 80% after five reuses. In the process of simulating the PAHs-contaminated site treatment experiment in Hangzhou Iron and steel plant, immobilized laccase exhibited efficient adsorption and degradation performances and even the removal rate of 5-ring PAHs reached more than 90% in 40 days, resulting in improving urease activity and dehydrogenase in the soil and promoted the growth of a PAH degrading bacteria (Massilia). Our results further explained the efficient degradation effects of Fe3O4@NaBC@GA@LC on PAHs, which make it a promising candidate for PAHs-contaminated soil remediation.

Design and Optimization of Laccase Immobilization in Cellulose Acetate Microfiltration Membrane for Micropollutant Remediation

The industrial and environmental applications of laccase, especially in wastewater treatment, have gained focus in recent years. Therefore, developing the proper laccase immobilization techniques, which could improve the stability of the enzymes and simplify the required downstream processes, is needed. A novel two-step immobilization process was developed, resulting in cross-linked enzyme aggregates (CLEA) in the pores of the membrane. Laccase adsorption on a biodegradable cellulose acetate microfiltration membrane along with cross-linking was investigated to maximize the enzyme load and immobilization efficiency. The optimization was done regarding the: pH, temperature, enzyme concentration, adsorption time, cross-linker concentration, and temperature. It was concluded that the highest immobilization efficiency (76%) could be achieved in acidic buffers at 29 °C with high surface activity (1174 U·m−2) at the cost of partial denaturation and membrane fouling. The membrane was successfully utilized for the enzymatic treatment of diclofenac, and 58% removal efficiency was achieved. The results indicated that cellulose acetate is a suitable carrier for adsorption-based immobilization of laccase for the potential for environmental utilisation.

Laccase multi-point covalent immobilization: characterization, kinetics, and its hydrophobicity applications.

Laccase from Myceliophthora thermophila was immobilized using one-point and multi-point covalent attachment on both a native and a modified new commercial epoxy carrier (Immobead 150P). After 10 cycles of operation at pH 3.0 and temperature 70°C, the multi-point covalently immobilized laccase on the modified Immobead 150P performed best in terms of immobilization characteristics, retaining 95% of its initial activity. Thermodynamic parameters of thermal inactivation emphasized the positive impact of the immobilization procedure. At 50°C, the immobilized and free enzyme activity levels dropped by 27 and 73%, respectively, after 48h of incubation. The immobilized enzyme enhanced its stability in alkaline conditions, resuming 95% of its original activity after 3h at pH 9.0. Immobilization reduced substrate affinity because the free laccase's Km value was lower than that of the immobilized laccase. Finally, the application of immobilized laccase in an innovative wood treatment process was tested by grafting lauryl gallate (LG) in order to provide hydrophobic properties to the wood. The results showed a relative water contact angle of 85.7% for treated wood, whereas the control showed only 26.6%, after 4min of contact between water and beechwood surface. KEY POINTS: • Multi-point covalent immobilization of a commercial laccase on a commercial support. • Enzymatic parameters generally improved by immobilization process. • New application of immobilized laccase: enzymatic-assisted wood hydrophobization.

Biodegradation of acid orange-7 dye by immobilized laccase on functionalized ZSM-5 zeolites: Investigation of the role of functionalization and SiO2/Al2O3 ratio of zeolite on the catalytic performance

It is known that SiO2/Al2O3 ratio and functionalization affect the features of zeolites. Considering these facts, in this study, two ZSM-5 zeolites with different SiO2/Al2O3 ratios of 200 and 80 (Z(80) and Z(200)) were synthesized and successively functionalized with (3-chloropropyl) triethoxysilane, melamine, and glutaraldehyde to furnish F-Z(80) and F-Z(200). The samples were utilized for the immobilization of laccase generating four biocatalysts, Lac@Z(80), Lac@Z(200), Lac@F-Z(80), and Lac@F-Z(200), able to biodegrade acid orange-7 dye. Comparison of the catalytic performance of the catalysts with free laccase indicated the remarkably improved performance of the supported laccase. Furthermore, Lac@F-Z(80) and Lac@F-Z(200) exhibited higher activity, recyclability, and storage stability compared to Lac@Z(80) and Lac@Z(200). Functionalization also enhanced the stability of biocatalysts over the change of pH and temperature. These observations were attributed to the role of the functional group in improvement of anchoring of laccase through covalent interactions. In the cases of Lac@Z(80) and Lac@Z(200), SiO2/Al2O3 ratio did not significantly affect the stability of the catalysts, while Lac@F-Z(80) was less susceptible to temperature and Lac@Z(200) showed higher pH stability. LC-MS analysis was also conducted and showed that the main compounds produced from biodegradation of acid orange-7 dye were 4-aminobenzenesulfonic acid, 2-amino-1-naphthol, and 1-amino-2-naphthol, 1,4-naphthoquinone, and 2-naphthol.

A Novel Approach, Based on the Combined Action of Chitosan Hydrogel and Laccases, for the Removal of Dyes from Textile Industry Wastewaters.

Dyes are considered as one the most important classes of contaminants that threaten the environment and human life. The synergy between the adsorption capacity of chitosan hydrogels and the catalytic properties of the enzyme laccase was exploited to improve the removal of contaminants from a liquid stream. The adsorption capacity of a chitosan hydrogel was tested on three different textile dyes. The effect of pH on the adsorption efficiency was dependent on the dye tested: the removal of methylene blue (MB), a cationic dye, was more effective at alkaline values of pH, whereas bromophenol blue (BPB) and Coomassie brilliant blue (BB), both anionic dyes, were more effectively removed under acid environments. The use of laccase immobilized onto chitosan has significantly improved the efficiency of dye removal, exploiting the synergy between the adsorption capacity of chitosan and the catalytic properties of the enzyme. The simultaneous processes of adsorption and enzymatic degradation improved the dye removal whatever the pH value adopted, making the removal efficiency less dependent from the pH changes. The chitosan used as a support for the immobilization of laccases showed good stability under repeated cycles, demonstrating the feasibility of the method developed for the application in wastewater remediation.

Immobilization of laccase on carboxyl-functionalized chitosan-coated magnetic nanoparticles with improved stability and reusability

Immobilization of laccase on carboxyl-functionalized chitosan-coated magnetic nanoparticles with improved stability and reusability

Immobilization of laccase on Fe3O4@MF-CNTs for the rapid and sensitive biosensing of catechol

An Fe3O4@MF-CNTs-Lac biosensor was developed for rapid and sensitive detection of catechol.

Lignocellulosic residues from bioethanol production: a novel source of biopolymers for laccase immobilization.

The full utilization of the main components in the lignocellulosic biomass is the major goal from a biorefinery point of view, giving not only environmental benefits but also making the process economically viable. In this sense the solid residue obtained in bioethanol production after steam explosion pretreatment, enzymatic hydrolysis, and fermentation of the lignocellulosic biomass, was studied for further valorization. Two different residues were analyzed, one generated by the production of cellulosic ethanol from an energy crop such as switchgrass (Panicum virgatum) and the other, from wood (Eucalyptus globulus). The chemical composition of these by-products showed that they were mainly composed of lignin with a total content range from 70 to 83% (w/w) and small amounts of cellulose and hemicellulose. The present work was focused on devising a new alternative for processing these materials, based on the ability of the ionic liquids (IL) to dissolve lignocellulosic biomass. The resulting mixture of biopolymers and IL constituted the raw material for developing new insoluble biocatalysts. Active hydrogels based on fungal laccase from Dichostereum sordulentum 1488 were attained. A multifactorial analysis of the main variables involved in the immobilization process enabled a more direct approach to improving hydrogel-bound activity. These hydrogels achieved a 97% reduction in the concentration of the estrogen ethinylestradiol, an emerging contaminant of particular concern due to its endocrine activity. The novel biocatalysts based on fungal laccase entrapped on a matrix made from a by-product of second-generation bioethanol production presents great potential for performing heterogeneous catalysis offering extra value to the ethanol biorefinery.

Improved stability and activity of laccase through de novo and post-synthesis immobilization on a hierarchically porous metal-organic framework (ZIF-8).

Porous materials such as metal-organic frameworks (MOFs) are considered to be suitable materials for immobilizing enzymes to improve their stability. However, conventional MOFs reduce the enzymes' catalytic activity due to difficulties with mass transfer and diffusing reactants after their micropores are occupied by enzyme molecules. To address these issues, a novel hierarchically structured zeolitic imidazolate framework-8 (HZIF-8) was prepared to study the effects of different laccase immobilization approaches such as the post-synthesis (LAC@HZIF-8-P) and de novo (LAC@HZIF-8-D) immobilization of catalytic activities for removing 2,4-dichlorophenol (2,4-DCP). The results showed higher catalytic activity for the laccase-immobilized LAC@HZIF-8 prepared using different methods than for the LAC@MZIF-8 sample, with 80% of 2,4-DCP removed under optimal conditions. These results could be attributable to the multistage structure of HZIF-8. The LAC@HZIF-8-D sample was stable and superior to LAC@HZIF-8-P, maintaining a 2,4-DCP removal efficiency of 80% after three recycles and demonstrating superior laccase thermostability and storage stability. Moreover, after loading with copper nanoparticles, the LAC@HZIF-8-D approach exhibited a 2,4-DCP removal efficiency of 95%, a promising finding for its potential use in environmental purification.

Laccase-Carrying Polylactic Acid Electrospun Fibers, Advantages and Limitations in Bio-Oxidation of Amines and Alcohols.

Laccases are oxidative enzymes that could be good candidates for the functionalization of biopolymers with several applications as biosensors for the determination of bioactive amine and alcohols, for bioremediation of industrial wastewater, and for greener catalysts in oxidation reactions in organic synthesis, especially used for non-phenolic compounds in combination with redox mediators in the so-called Laccase Mediator System (LMS). In this work, we describe the immobilization of Laccase from Trametes versicolor (LTv) in poly-L-lactic acid (PLLA) nanofibers and its application in LMS oxidation reactions. The PLLA-LTv catalysts were successfully produced by electrospinning of a water-in-oil emulsion with an optimized method. Different enzyme loadings (1.6, 3.2, and 5.1% w/w) were explored, and the obtained mats were thoroughly characterized. The actual amount of the enzyme in the fibers and the eventual enzyme leaching in different solvents were evaluated. Finally, the PLLA-LTv mats were successfully applied as such in the oxidation reaction of catechol, and in the LMS method with TEMPO as mediator in the oxidation of amines with the advantage of easier work-up procedures by the immobilized enzyme. However, the PLLA-LTv failed the oxidation of alcohols with respect to the free enzyme. A tentative explanation was provided.

Laccase immobilization on hierarchical micro/nano porous chitin/graphene oxide beads prepared via Pickering emulsion template for dye decolorization

Nowadays, the removal of dye molecules from water sources is a major environmental concern and challenge. Enzymatic degradation (biological) and adsorptive (physical) dye removal are considered to be the most effective dye removal techniques. Laccase catalysis is a reliable method to remove synthetic dyes from water to reduce environmental pollution, but free laccase has the disadvantages of low stability and non-reusability. In this study, stable Pickering emulsions were prepared using chitin (Ch) and graphene oxide (GO) composite solutions. Ch/GO composite aerogel beads with hierarchical porous structure were obtained by emulsion template method combined with freeze-drying, and then used for laccase immobilization and dye decolorization experiments. The results showed that the immobilized laccase had better thermal stability compared to the free enzyme, and it was found that the optimum pH of the immobilized laccase was 3.4 and the optimum temperature was 55 °C. In the cyclic decolorization experiments of acid Big Red GR, the decolorization rate of CH/GO-30 was maintained at 100% after 5 cycles of ABTS (2,2-biazo-bis(3-ethyl-benzothiazole-6-sulfonic acid) diammonium salt) by enzyme catalysis and carrier adsorption synergistically. The preparation of Ch/GO composite aerogel beads by the Pickering emulsion template method is simple and green, without the use of surfactants, avoiding secondary pollution to the environment, and has potential industrial applications in the decolorization and biodegradation of dye wastewater.

Laccase–copper phosphate hybrid nanoflower as potent thiazole remediation agent

The increase in the global population in recent years has caused a fast increase in pollution problems worldwide. One of these problems is emerging pollutants, which are a class of human-made organic compounds that have been detected in various water bodies. To develop a recyclable, reusable, and efficient material for the degradation of these pollutants, we report the synthesis, characterization and application of hybrid nanoflowers imbedded with laccase enzymes. These copper phosphate-laccase nanomaterials had a spherical flower-like shape, a surface area of 53.46 m2/g, and a mean pore size of 10.58 nm. When tested against a panel of diverse compounds, the hybrid nanoflowers were able to degrade a number of emerging pollutants, including 2-mercaptobenzothiazole (MBT), caffeic acid, thiabendazole, cimetidine, and sulfamethoxazole, with efficiencies of 97 %, 97.0 %, 78.1 %, 72.3 %, and 51.3 %, respectively. For 2-mercaptobenzothiazole (MBT), we identified the intermediate produced during the degradation process. In addition, the nanoflowers were able to degrade >90 % MBT in a spiked ground water sample. Furthermore, we tested the recyclability and storage stability of the hNFs for the degradation of MBT and found that they could be reused for five cycles and stored for 21 days at 4 °C. Our results suggest that laccase-embedded hybrid nanoflowers to be potent remediation agents for the degradation of thiazole as well as other emerging pollutants. Also, the successful immobilization of laccase on hybrid nanoflowers shown in the present study can allow for the efficient recycling of laccase enzymes for multiple degradation cycles leading to potential scaling up and building of a bioreactor.

Laccase immobilization on bimetallic MOF‐derived porous carbon materials for the removal of bisphenol A

AbstractAs a result of the poor mechanical properties of most metal–organic frameworks (MOFs), they are liable to dissociate in water and thus cannot maintain immobilized enzyme activity. In this paper, a magnetic porous carbon material (Fe‐Cu‐MPC) with good stability was prepared by high temperature carbonization using a bimetallic Fe‐Cu‐MOF precursor, and used to immobilize laccase by a simple and rapid adsorption‐crosslinking method. Optimized by the response surface method, optimal conditions for the immobilization of laccase on Fe‐Cu‐MPC were obtained as follows: enzyme concentration 1.275 mg mL−1, immobilization time 32 min, and pH 4.7. Under the optimal preparation conditions, the activity of the immobilized laccase obtained was 0.63 U mg−1, with activity recovery of 63.13%. The enzyme loading was 189.03 mg g−1 and the immobilization yield was 74.13%. Compared with free laccase, Fe‐Cu‐MPC@Lac showed stronger tolerance to temperature and pH, and had better thermal stability and storage stability. The removal of BPA by immobilized laccase was studied, showing that the removal effect of Fe‐Cu‐MPC@Lac on BPA was much higher than that of free laccase. Within 5 h, the BPA removal amount of Fe‐Cu‐MPC@Lac (0.34 mmol g−1) was two‐fold that of free laccase (0.14 mmol g−1), and the BPA removal rate reached 41.35% after ten cycles. Finally, based on the intermediates identified by gas chromatography–mass spectrometry, two possible degradation pathways of BPA were proposed. Studies have shown that Fe‐Cu‐MPC is an excellent carrier and Fe‐Cu‐MPC@Lac has great potential in the treatment of BPA containing wastewater. © 2022 Society of Chemical Industry (SCI).

Sandwich-like heterostructured nanomaterials immobilized laccase for the degradation of phenolic pollutants and boosted enzyme stability

A novel magnetic 2D/2D heterogeneous structure MXene@NiFe-LDH@Fe3O4 was prepared for immobilization of laccase. In this work, two-dimensional MXene nanosheets with abundant surface functional groups were heterogeneously assembled with layered double hydroxide (LDH) by in situ co-precipitation method, and magnetic nanoparticle Fe3O4 with excellent biocompatibility and rapid separation of materials and substrates was introduced subsequently, and then silane coupling agent was coated on the surface of MXene@NiFe-LDH@Fe3O4. The functionalized MXene@NiFe-LDH@Fe3O4 was employed as a carrier to immobilize laccase from Trametes-Versicolor. The enzyme loading of the nanocomposite material is as high as 167.9 mg/g. Compared with free enzymes, the immobilized laccase showed a notable improvement in stability in a wider range of pHs (2.0–8.0), temperatures (25–60 °C), and organic solvent concentration (1–5 M). The reusability study suggested that after 7 cycles of repeated catalysis, the degradation efficiency could reach 55.5% for 2,4-dichlorophenol, 92.1% for bisphenol A and70.9% for pyrocatechol. The results provide a new carrier preparation strategy for the efficient immobilization of laccase.