Immobilization Of Peroxidase Research Articles

Construction of hierarchical porous MIL-100(Fe) for horseradish peroxidase immobilization and its degradation performance of bisphenol A

Construction of hierarchical porous MIL-100(Fe) for horseradish peroxidase immobilization and its degradation performance of bisphenol A

Enhanced Immobilization of Enzymes on Plasma Micro-Nanotextured Surfaces and Microfluidics: Application to HRP

The enhanced and direct immobilization of the enzyme horseradish peroxidase on poly(methyl methacrylate) (PMMA) microchannel surfaces to create a miniaturized enzymatic reactor for the biocatalytic oxidation of phenols is demonstrated. Enzyme immobilization occurs by physical adsorption after oxygen plasma treatment, which micro-nanotextures the PMMA surfaces. A five-fold enhancement in immobilized enzyme activity was observed, attributed to the increased surface area and, therefore, to a higher quantity of immobilized enzymes compared to an untreated PMMA surface. The enzymatic reaction yield reached 75% using a flow rate of 2.0 μL/min for the reaction mixture. Additionally, the developed microreactor was reused more than 16 times without affecting the enzymatic conversion yield. These results demonstrate the potential of microchannels with plasma micro/nanotextured surfaces for the rapid and facile fabrication of microfluidic enzymatic microreactors with enhanced catalytic activity and stability.

Fabrication of hierarchical 3D layered yttrium hydroxide nanocomposites: Their use for enzyme immobilization and catalysis of H2O2

3D layer metal hydroxides (LMHs) have attracted significant interest due to their distinctive structural features. Unfortunately, the limited electrical conductivity hinders the potential utilization of LMHs in electrochemical analysis. To address this issue, herein, gold nanoparticles (AuNPs) were introduced to layered yttrium hydroxides by a facile in-situ electrodeposition method. Subsequently, the hierarchical AuNPs/LYH nanocomposite was employed as solid supports for immobilization of horseradish peroxidase, generating an enzymatic biosensor for hydrogen peroxide detection. As a result, a linear range of 1–500 μM with a remarkably low detection limit of 0.33 μM (S/N=3) was achieved. Moreover, the biosensor also demonstrated excellent selectivity, reproducibility and stability. The superior electrochemical responses of the HRP/AuNPs/LYH nanocomposite towards H2O2 can be ascribed to the synergistic effect arising from the flower-like structures of LYHs and the high conductivity of AuNPs. Our study provides a significant approach for the development of enzyme immobilized supports through the construction of layer metal hydroxide nanocomposites.

Phenol Removal through Horseradish Peroxidase Immobilization on Ultrafiltration Membranes: Comparative Analysis of Immobilization Methods and Fouling Patterns

This research investigates the removal of phenol using pure peroxidase from horseradish grade I in conjunction with a dead-end ultrafiltration membrane. Various horseradish peroxidase (HRP) immobilization techniques— physical adsorption, covalent bonding, and cross-linking with glutaraldehyde—were applied to a regenerated cellulose (RC) membrane with a surface area of 44 m2 and a molecular weight cut-off of 30 kDa. The investigation examined factors influencing phenol removal, including phenol concentration, membrane fouling, and the reusability of immobilized enzymes. Results indicated that covalent bonding was the most suitable enzyme immobilization technique, achieving a remarkable 90.1% immobilization yield. Phenol removal efficiency reached 100% at 30 min under specific conditions: phenol concentration of 1 mg/L, pH 6.0, hydrogen peroxide concentration of 0.5 mM, and operating pressure set at 3 psig, with temperature maintained at 28 ± 3 °C. Membrane fouling resulted in a decrease in flux. The performance of fouling models was found to be influenced by phenol concentration, with the Cake Formation Model (CFM) proving most effective at low concentrations, while the Complete Pore Blocking Model (CBM) emerged as more suitable at higher concentrations. The immobilized enzyme exhibited reusability for five cycles, maintaining a phenol removal efficiency exceeding 50%. These findings contribute to understanding the enzymatic phenol removal process and the use of appropriate enzyme immobilization techniques for the effective and sustainable treatment of phenol-contaminated water.

Increasing Multienzyme Cascade Efficiency and Stability of MOF via Partitioning Immobilization.

Enhancing the stability of multienzyme cascade reactions in metal-organic frameworks (MOFs) is a challenging task in the fields of biotechnology and chemistry. However, addressing this challenge could yield far-reaching benefits across the application range in the biomedical, food, and environmental sectors. In this study, multienzyme partitioning immobilization that sequentially immobilizes cascade enzymes with hierarchical MOFs is proposed to reduce substrate diffusion resistance. Conversion results of ginsenosides indicate that this strategy improves the cascade efficiency up to 1.26 times. The substrate diffusion model is used to investigate the dual-interenzyme mass transfer behavior of substrates in the restricted domain space and evaluate the substrate channeling effect under partitioning immobilization. Molecular docking and kinetic simulations reveal that the MOFs effectively limit the conformational changes of cascade enzymes at high temperatures and in organic solvents while maintaining a large pocket of active centers. This phenomenon increased efficient substrate docking to the enzyme molecules, further optimizing cascade efficiency. The results of the immobilization of GOX and horseradish peroxidase as model enzymes indicate that the partitioned MOF immobilization strategy could be used for universal adaptation of cascade enzymes.

Smartphone-assisted portable paper-based biosensors for rapid and sensitive detection of biomarkers in urine

In the increasing demand for real-time testing, the imperative for a straightforward, rapid, portable, and effective biosensor is evident. In this study, an intelligent paper-based biosensor was developed, employing cost-effective and readily available filter paper as a matrix to prepare boronate affinity paper-based materials. The immobilization of horseradish peroxidase (HRP) from crude horseradish extract onto the material surface was realized. Ultimately, a cold assembly technique was employed to construct this intelligent paper-based biosensor, which combined with a smartphone APP, facilitated the easy, accurate, and simultaneous detection of the content of glucose (GLU), uric acid (UA), and sarcosine (SAR) in urine, significantly enhancing the convenience and practicality of real-time monitoring. Additionally, UV spectroscopy served as an auxiliary method to affirm the reliability of the detection outcomes. For the detection of GLU, UA, and SAR, the biosensor exhibited linear ranges of 2–200, 2–500, and 2–500 µmol L–1, respectively, with detection limits (LODs) of 1 µmol L–1 for all analytes. In practical urine sample analysis, the recovery rates of GLU, UA, and SAR detected by the intelligent paper-based biosensors were within a reasonable range, with relative standard deviations below 3.5 %. This fully demonstrates the high accuracy and reliability of the intelligent paper-based biosensor. The development of this low-cost, portable biosensor provides a new method for the detection of biomarkers related to H2O2.

Acrylic modification as an environmentally acceptable supporter for improving peroxidase enzyme: stability and reusability

This study focuses on the immobilization of horseradish peroxidase (HRP) on modified acrylic fabrics incorporating gold and silver nanoparticles. The process involves treating the acrylic fabrics with hydroxylamine hydrochloride and then coating them with silver and gold nanoparticles. Both obtained materials, treated acrylic fabrics-coated with silver nanoparticles (AgNPs@TA-HAC) and treated acrylic fabrics-coated with gold nanoparticles (AuNPs@TA-HAC), were utilized as supporters for HRP. The physicochemical properties of modified acrylic fabrics were investigated using FTIR, SEM, and zeta potential. HRP immobilized on AgNPs@TA-HAC displayed an activity of 69 units/g support with a specific activity of 4.55 units/mg protein, whereas HRP immobilized on AuNPs@TA-HAC demonstrated an activity of 76 units/g support with a specific activity of 4.75 units/mg protein. After the 15 repetitive cycles, the immobilized HRP on AuNPs@TA-HAC and AgNPs@TA-HAC retained 75 and 59% of their enzymatic activity, respectively. The immobilized HRP on both material supporters retained its activity better compared to the free HRP when exposed to all tested organic solvents. The Michaelis constant (Km) for free HRP and HRP immobilized on AuNPs@TA-HAC and AgNPs@TA-HAC were determined to be 6.23, 8.65, and 9.11 mM, respectively. The maximum reaction rates (V max) for the immobilized HRP on both supports were slightly reduced at 0.71 and 0.69 U/mL, compared to 0.74 U/mL for free HRP. This approach of utilizing acrylic fabrics and metal nanoparticles provides a promising method for enzyme immobilization, with potential applications in various industries.

Sweat lactate biosensor based on lactate oxidase immobilized with flower-like NiCo2O4 and carbon nanotubes

We design a portable and non-invasive lactate biosensor that can conveniently monitor lactate levels in real human sweat. The proposed biosensor is constructed through the immobilization of lactate oxidase (LOx) and horseradish peroxidase (HRP) on flower-like NiCo2O4 microspheres coupled with single-walled carbon nanotubes (SWCNTs). The synergistic effect between NiCo2O4 microspheres and SWCNTs can increase the conductivity and the active surface area of the electrode. Using ferrocene–methanol (FcMeOH) as the electron transfer mediator in buffer, the developed biosensor can detect lactate in a wide linear range of 0.1 mM to 30 mM with a detection limit of 39.9 µM under optimal conditions. Additionally, the biosensor exhibits excellent selectivity and stability. As proof of concept, the biosensor was applied to monitor lactate level in real human sweat samples, and the results were in good agreement with those obtained by high-performance liquid chromatography (HPLC) analysis. The lactate biosensor is cost effective and can be as a non-invasive alternative to blood analysis, which is suitable for rapid analysis of lactate levels in the fields of fitness, health monitoring and clinical diagnosis.

Immobilization of Horseradish Peroxidase and Myoglobin Using Sodium Alginate for Treating Organic Pollutants

Removing organic pollutants from wastewater is crucial to prevent environmental contamination and protect human health. Immobilized enzymes are increasingly being explored for wastewater treatment due to their specific catalytic activities, reusability, and stability under various environmental conditions. Peroxidases, such as horseradish peroxidase (HRP) and myoglobin (Mb), are promising candidates for immobilized enzymes utilized in wastewater treatment due to their ability to facilitate the oxidation process of a wide range of organic molecules. However, the properties of the carrier and support materials greatly influence the stability and activity of immobilized HRP and Mb. In this research, we developed immobilized HRP and Mb using support material composed of sodium alginate and CaCl2 as carriers and glutaraldehyde as a crosslinking agent. Following this, the efficacy of immobilized HRP and Mb in removing aniline, phenol, and p-nitrophenol was assessed. Both immobilized enzymes removed all three organic pollutants from an aqueous solution, but Mb was more effective than HRP. After being immobilized, both enzymes became more resilient to changes in temperature and pH. Both immobilized enzymes retained their ability to eliminate organic pollutants through eight treatment cycles. Our study uncovered novel immobilized enzyme microspheres and demonstrated their successful application in wastewater treatment, paving the way for future research.

Highly Light-Harvesting MOF-on-MOF Heterostructure: Cascading Functionality to Flexible Photogating of Organic Photoelectrochemical Transistor and Bienzyme Cascade Detection.

Recently, organic photoelectrochemical transistor (OPECT) bioanalysis has become a prominent technique for the high-performance detection of biomolecules. However, as a sensitive index of the OPECT, the dynamic regulation transconductance (gm) is still severely deficient. Herein, this work reports a new photosensitive metal-organic framework (MOF-on-MOF) heterostructure for the effective modulation of maximum gm and natural bienzyme interfacing toward choline detection. Specifically, the bidentate ligand MOF (b-MOF) was assembled onto the UiO-66 MOF (u-MOF) by a modular assembly method, which could facilitate the charge separation and generate enhanced photocurrents and offer a biophilic environment for the immobilization of choline oxidase (ChOx) and horseradish peroxidase (HRP) through hydrogen-bonded bridges. The transconductance of the OPECT could be flexibly altered by increased light intensity to maximal value at zero gate bias, and sensitive choline detection was achieved with a detection limit of 0.2 μM. This work reveals the potential of MOF-on-MOF heterostructures for futuristic optobioelectronics.

Determination of the total antioxidant capacity of the Chinese tea based on a novel "peroxidase/zirconium phosphonate"composite electrochemical sensor.

In this work, a novel zirconium phosphonate (ZrPR1R2) was prepared by decorating both the aminoethoxy- group (R1) and the carboxypropyl- group (R2) on the zirconium phosphate layers in order to manipulate further the immobilization of the peroxidase (POD), and an antioxidant biosensor with higher sensitivity was constructed by dropping the POD/ZrPR1R2 composite onto the glassy carbon electrode surface. The activity of the POD/ZrPR1R2 composite was detected by Uv-vis spectra. The direct electrochemical behavior, the electrocatalytic response to dissolved oxygen and hydrogen peroxide, as well as the ability to detect total antioxidant capacity in tea sample were investigated by the methods of cyclic voltammetry. The results indicated that the immobilization of POD in ZrPR1R2 nanosheets matrix enhanced the enzymatic activity, and achieved the fast and direct electron transfer between POD and glassy carbon electrode. Moreover, the POD/ZrPR1R2 composite modified electrode show the electrocatalytic response to hydrogen peroxide in the linear range of 8.8×10-8 to 8.8×10-7 mol L-1, with the detection limit of 3.3×10-8 mol L-1. Attributing to the sensitive response to dissolved oxygen, the total antioxidant capacity can be detected directly in the real tea water by this POD/ZrPR1R2 composite modified electrode.

Support Materials of Organic and Inorganic Origin as Platforms for Horseradish Peroxidase Immobilization: Comparison Study for High Stability and Activity Recovery.

In the presented study, a variety of hybrid and single nanomaterials of various origins were tested as novel platforms for horseradish peroxidase immobilization. A thorough characterization was performed to establish the suitability of the support materials for immobilization, as well as the activity and stability retention of the biocatalysts, which were analyzed and discussed. The physicochemical characterization of the obtained systems proved successful enzyme deposition on all the presented materials. The immobilization of horseradish peroxidase on all the tested supports occurred with an efficiency above 70%. However, for multi-walled carbon nanotubes and hybrids made of chitosan, magnetic nanoparticles, and selenium ions, it reached up to 90%. For these materials, the immobilization yield exceeded 80%, resulting in high amounts of immobilized enzymes. The produced system showed the same optimal pH and temperature conditions as free enzymes; however, over a wider range of conditions, the immobilized enzymes showed activity of over 50%. Finally, a reusability study and storage stability tests showed that horseradish peroxidase immobilized on a hybrid made of chitosan, magnetic nanoparticles, and selenium ions retained around 80% of its initial activity after 10 repeated catalytic cycles and after 20 days of storage. Of all the tested materials, the most favorable for immobilization was the above-mentioned chitosan-based hybrid material. The selenium additive present in the discussed material gives it supplementary properties that increase the immobilization yield of the enzyme and improve enzyme stability. The obtained results confirm the applicability of these nanomaterials as useful platforms for enzyme immobilization in the contemplation of the structural stability of an enzyme and the high catalytic activity of fabricated biocatalysts.

Development of chia gum/alginate-polymer support for horseradish peroxidase immobilization and its application in phenolic removal

Chia gum’s molecular structure with distinctive properties as well as the alginate-based hydrogel’s three-dimensionally cross-linked structure can provide a potent matrix for immobilization of enzyme. Herein, chia gum (CG)/alginate (A)-polymeric complex was synthesized and employed as a support material for the immobilization of horseradish peroxidase (HRP). HRP was successfully immobilized on the developed ACG-polymeric support, and the highest immobilization recovery (75%) was observed at 1.0% CG and 2% A, pH 7.0, and 50 units of the enzyme. The structure, morphology, and thermal properties of the prepared ACG-HRP were demonstrated using Fourier Transform Infrared (FTIR), Scanning Electron Microscope, and Thermogravimetric (TGA) analyses. ACG-HRP showed a good reusability (60%) over ten reuses. The immobilized ACG-HRP displayed an acidic pH optimum (6.0), a higher temperature optimum (50 °C), and improved thermal stability (30–50 °C) compared to the soluble HRP at pH 7.0, 40 °C and (30–40 °C), respectively. ACG-HRP has a lower affinity for hydrogen peroxide (H2O2) and guaiacol and a higher oxidizing affinity for a number of phenolic substrates. The ACG-HRP demonstrated greater resistance to heavy metals, isopropanol, urea, Triton X-100, and urea, as well as improved efficiency for eliminating phenol and p-chlorophenol. The developed ACG-polymeric support provided improved enzyme properties, allowed the reuse of the immobilized HRP in 10 cycles, and made it promising for several biotechnological applications.

Immobilization of periodate-oxidized horseradish peroxidase by adsorption on sepiolite

Horseradish peroxidases (HRP), native and periodate-oxidized were immobilized onto sepiolite clay mineral by adsorption. Both peroxidases were adsorbed on this carrier in different quantities. Specific activity of immobilized enzymes was increased with increasing the amount of peroxidase added per gram of sepiolite. The highest specific activity was achieved when 15 mg of peroxidase was added per gram of sepiolite. Also, periodate-oxidized enzymes showed similar specific activity as native ones. Stability studies (pH, thermal and operational stability) were conducted for both peroxidases. Residual specific activity of HRP immobilized onto sepiolite declined with an increase of incubation time at 65?C. Oxidized-peroxidase lost 64 % of the initial activity, whereas native HRP dropped 92 % of its activity after 5 minutes of incubation at 65?C. Reduction of the enzyme activity was observed with the temperature increase from 30 to 80 ?C. pH profiles of native peroxidase immobilized onto sepiolite were higher in both acidic and basic regions compared to periodate-oxidized enzyme. Oxidized HRP was more successful in studies of operational stability, it retained 42 % of its activity after 4 consecutive cycles of pyrogallol oxidation, whereas native peroxidase kept only 11 % of the original activity. Keywords: modified enzyme; specific activity; thermostability; reusability.

Immobilization of Horseradish Peroxidase on Ca Alginate-Starch Hybrid Support: Biocatalytic Properties and Application in Biodegradation of Phenol Red Dye.

In this study, horseradish peroxidase was extracted, purified, and immobilized on a calcium alginate-starch hybrid support by covalent bonding and entrapment. The immobilized HRP was used for the biodegradation of phenol red dye. A 3.74-fold purification was observed after precipitation with ammonium sulfate and dialysis. An immobilization yield of 88.33%, efficiency of 56.89%, and activity recovery of 50.26% were found. The optimum pH and temperature values for immobilized and free HRP were 5.0 and 50°C and 6.5 and 60°C, respectively. The immobilized HRP showed better thermal stability than its free form, resulting in a considerable increase in half-life time (t1/2) and deactivation energy (Ed). The immobilized HRP maintained 93.71% of its initial activity after 45days of storage at 4°C. Regarding the biodegradation of phenol red, immobilized HRP resulted in 63.57% degradation after 90min. After 10 cycles of reuse, the immobilized HRP was able to maintain 43.06% of its initial biodegradative capacity and 42.36% of its enzymatic activity. At the end of 15 application cycles, a biodegradation rate of 8.34% was observed. In conclusion, the results demonstrate that the immobilized HRP is a promising option for use as an industrial biocatalyst in various biotechnological applications.

A sensitive biosensor for cyanide detection using modified glassy carbon electrode with reduced graphene oxide and immobilization of horseradish peroxidase

AbstractCyanide is a highly poisonous and hazardous substance which may release into the environment from natural sources or industrial effluent; therefore, cyanide detection is a fundamental step to prevent environmental pollution and secure health and safety. In this study, we prepared a sensitive amperometric inhibition biosensor for cyanide detection by immobilization of horseradish peroxidase (HRP) enzyme and reduced graphene oxide (rGO) on the surface of glassy carbon electrode (GCE). To do so, we performed the amperometric measurement by modified GCE to test its efficiency in detecting cyanide. The optimum conditions of pH equal to 7.5, −100 mV applied potential, 0.7 μM mediator concentration, and 0.5 mM substrate concentration were found. Then, experiments were performed at different boundary conditions in a range of 0.1 to 10 μM cyanide concentration at optimal conditions and a low detection limit of 0.01 μM was obtained. Also, the possible mechanism of inhibition was analyzed based on the Michalis–Menten equation and non‐competitive inhibition was observed. Due to high sensitivity, low detection limit, and low cost, this biosensor is proposed as a useful method for cyanide determination in real samples.

Immobilization of horseradish peroxidase on metal-organic framework to improve enzyme activity for enhanced chemodynamic therapy

Bio-enzymes have the advantages of strong substrate specificity, high catalytic efficiency, and minimal toxic side effects, making them promising drugs in cancer therapy. However, the poor stability and cellular penetrability of uncoated protein in the physiological environment severely restricts the direct application of Bio-enzyme. To address it, we report a metal-organic framework (MOF), Hf-DBA (H2DBA, biphenyl carboxylic acid ligands). The morphology of the Hf-DBA was revealed by TEM and the diameter was in the range of 200 to 350 nm. Hf-DBA acted a carrier for intracellular delivery and protection of horseradish peroxidase (HRP). The prepared HRP@Hf-DBA can catalyze the excess H2O2 in the tumor cells to generation of •OH for chemodynamic therapy (CDT). Compared with free HRP, the catalytic activity of HRP@Hf-DBA is significantly improved, and the optimal catalytic conditions are explored. The catalytic stability of HRP@Hf-DBA remained above 70% after 12 cycles of catalysis. After treatment with HRP@Hf-DBA, the apoptosis rates of A549 and Hela cells was 71.64%, and 76.86%. The results in vitro show that HRP@Hf-DBA can effectively inhibit the growth of tumor cells through enhanced CDT.

Efficient degradation of various recalcitrant azo dyes in aqueous medium by immobilized Origanum vulgare peroxidase

Hybrid xerogels, constructed from organic and inorganic silanes, have emerged as materials with versatile applications. They have shown promising potential for immobilization as their chemical structures encompass the mechanical, thermal, and structural stability of inorganic compounds in synergy with the functionality and flexibility of organic compounds. Incorporating a magnetic core and preparation at the nanoscale multiply their attraction for enzyme immobilization. To evaluate how efficiently this technology works for the immobilization of a plant peroxidase, a novel peroxidase (POXov) from a famous medicinal herb, Origanum vulgare, was immobilized on magnetic nanoparticles of a hybrid xerogel obtained from tetraethyl orthosilicate and (3-aminopropyl)triethoxysilane. Immobilization boosted the physicochemical properties of the enzyme so that the immobilized POXov (I.POXov) could tolerate lower pHs and higher temperatures. It oxidized all types of the examined phenolic, catecholic, guaiacolic, and aniline diazo dyes, while the free POXov (F.POXov) failed to oxidize the last group. These improvements also made I.POXov capable of oxidizing the recalcitrant azo dyes carrying electron-withdrawing groups much faster than F.POXov. I.POXov was employed in 7 successive cycles of oxidation of both phenolic and guaiacolic dyes with an average efficiency of 69%. The efficacy of the immobilization method and I.POXov competency for the enzymatic remediation of polluted water resources have been quantitatively demonstrated.

Enzyme-immobilized hierarchically porous covalent organic framework biocomposite for catalytic degradation of broad-range emerging pollutants in water

Efficient enzyme immobilization is crucial for the successful commercialization of large-scale enzymatic water treatment. However, issues such as lack of high enzyme loading coupled with enzyme leaching present challenges for the widespread adoption of immobilized enzyme systems. The present study describes the development and bioremediation application of an enzyme biocomposite employing a cationic macrocycle-based covalent organic framework (COF) with hierarchical porosity for the immobilization of horseradish peroxidase (HRP). The intrinsic hierarchical porous features of the azacalix[4]arene-based COF (ACA-COF) allowed for a maximum HRP loading capacity of 0.76 mg/mg COF with low enzyme leaching (<5.0 %). The biocomposite, HRP@ACA-COF, exhibited exceptional thermal stability (∼200 % higher relative activity than the free enzyme), and maintained ∼60 % enzyme activity after five cycles. LCMSMS analyses confirmed that the HRP@ACA-COF system was able to achieve > 99 % degradation of seven diverse types of emerging pollutants (2-mercaptobenzothiazole, paracetamol, caffeic acid, methylparaben, furosemide, sulfamethoxazole, and salicylic acid)in under an hour. The described enzyme-COF system offers promise for efficient wastewater bioremediation applications.

Immobilization of Horseradish Peroxidase for Phenol Degradation.

The use of enzymes to degrade environmental pollutants has received wide attention as an emerging green approach. Horseradish peroxidase (HRP) can efficiently catalyze the degradation of phenol in the environment; however, free HRP exhibits poor stability and temperature sensitivity and is easily deactivated, which limit its practical applications. In this study, to improve their thermal stability, HRP enzymes were immobilized on mesoporous molecular sieves (Al-MCM-41). Specifically, Al-MCM-41(W) and Al-MCM-41(H) were prepared by modifying the mesoporous molecular sieve Al-MCM-41 with glutaraldehyde and epichlorohydrin, respectively, and used as carriers to immobilize HRP on their surface, by covalent linkage, to form the immobilized enzymes HRP@Al-MCM-41(W) and HRP@Al-MCM-41(H). Notably, the maximum reaction rate of HRP@Al-MCM-41(H) was increased from 2.886 × 105 (free enzyme) to 5.896 × 105 U/min-1, and its half-life at 50 °C was increased from 745.17 to 1968.02 min; the thermal stability of the immobilized enzyme was also significantly improved. In addition, we elucidated the mechanism of phenol degradation by HRP, which provides a basis for the application of this enzyme to phenol degradation.