Encapsulation of HRP Enzyme onto a Magnetic Fe3O4 Np–PMMA Film via Casting with Sustainable Biocatalytic Activity

Abstract

Horseradish peroxidase (HRP) enzyme was effectively encapsulated onto an Fe3O4 nanoparticle–polymethyl methacrylate (PMMA) film via the casting method. The HRP was immobilized on the 0.5% Fe3O4Np–PMMA film and characterized by Fourier transform infrared spectroscopy and field emission scanning electron microscopy. Moreover, the reusability, thermal stability, optimum pH, optimum temperature, the influence of metal ions, and the effects of detergent and organic solvent were investigated. After optimizing the immobilization conditions, the highest efficiency of the immobilized enzyme was 88.4% using 0.5% Fe3O4Np–PMMA. The reusability of the immobilized HRP activity was 78.5% of its initial activity after being repeatedly used for 10 cycles. When comparing the free and immobilized forms of the HRP enzyme, changes in the optimum temperature and optimum pH from 30 to 40 °C and 7.0 to 7.5, respectively, were observed. The Km and Vmax for the immobilized HRP were estimated to be 41 mM, 0.89 U/mL for guaiacol and 5.84 mM, 0.66 U/mL for H2O2, respectively. The high stability of the immobilized HRP enzyme was obtained using metal ions, a high urea concentration, isopropanol, and Triton X-100. In conclusion, the applicability of immobilized HRP involves the removal of phenol in the presence of hydrogen peroxide, therefore, it could be a potential catalyst for the removal of wastewater aromatic pollutants.

Preparation of Streptavidin-Coated Magnetic Nanoparticles for Specific Immobilization of Enzymes with High Activity and Enhanced Stability

“…When α-amylase was immobilized following the same procedure as pullulanase, a similar shift of optimal temperature from 35 to 40 °C was observed (Figure S1). Also, similar improvements in thermal stability have been previously reported for various immobilized enzymes. , The change in optimal temperature observed here demonstrated that immobilized pullulanase was resistant to temperature increases, probably due to the immobilization-associated modifications to the physicochemical properties of the enzyme . Thus, with respect to industrial applications, immobilized enzymes are preferable to soluble forms, as the immobilized form can withstand the high temperatures that are important for enzyme catalysis …”

B,N-Doped PdRu Aerogels as High-Performance Peroxidase Mimics for Sensitive Detection of Glucose

“…45 As a comparison, the K m value with TMB of the B, N-PdRu aerogels was similar to that of horseradish peroxidase (HRP) and the K m of H 2 O 2 was nearly 100 times lower than the corresponding value of HRP, suggesting that B, N-PdRu aerogels have a better affinity for H 2 O 2 and TMB, as shown in Table 1. 46 As compared with some other reported nanozymes, the B, N-PdRu aerogels have a more superior peroxidase-like property.…”

Surface-coated magnetic nanostructured materials for robust bio-catalysis and biomedical applications-A review

“… Efficient removal of phenol in the presence of hydrogen peroxide. Removal of wastewater aromatic pollutants [123] Fe 3 O 4 –graphene nanocomposite Trametes Versicolor laccase APTES Stability and activity of the immobilized nanobiocatalyst was markedly increased than that to free laccase. Retained about 70% of its relative activity after incubating at 55 °C for 2 h, while only 48% of activity was recorded by the free laccase under identical time duration.…”