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Immobilization of Horseradish Peroxidase on Macroporous Glycidyl-Based Copolymers with Different Surface Characteristics for the Removal of Phenol

Journal of Polymers and the Environment volume 30, pages 3005–3020 (2022)Cite this article

Abstract

Novel macroporous copolymers of glycidyl methacrylate and ethylene glycol dimethacrylate with mean pore size diameters ranging from 150 to 310 nm were synthesized by dispersion polymerization and modified with ethylenediamine. The glutaraldehyde and periodate method were employed to immobilize horseradish peroxidase (HRP) onto these carriers. The activity of the immobilized enzyme was greatly affected by the pore size of the carrier. The highest specific activities of 9.65 and 8.94 U/g of dry weight were obtained for HRP immobilized by the periodate-route onto poly(GMA‐co‐EGDMA) carriers with pore size diameters of 234 and 297 nm, respectively. Stability studies showed an improved operational stability of immobilized peroxidase at 65 °C and in an organic solvent. HRP immobilized on a copolymer with a pore size of 234 nm, showing the highest specific activity and good stability, had higher activities at almost all pH values than the native enzyme and the increased Km value for pyrogallol oxidation. Immobilized HRP retained 80% of its original activity after five consecutive cycles of the pyrogallol oxidation and 98% of its initial activity in a storage stability study. Enzyme immobilized onto the macroporous copolymer with the pore size diameter of 234 nm showed a substantial degree of phenol removal achieved by immobilized peroxidase.

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Funding

This work was financially supported by the Ministry of Education, Science and Technological Development of the Republic of Serbia [Grant No. 451-03-9/2021-14/200168, University of Belgrade-Faculty of Chemistry; Grant No. 451-03-9/2021-14/200288, University of Belgrade-Innovation Center of the Faculty of Chemistry and Grant No. 451-03-9/2021-14/200053, University of Belgrade-Institute for Multidisciplinary Research].

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Authors and Affiliations

  1. University of Belgrade, Institute for Multidisciplinary Research, Kneza Višeslava 1, 11030, Belgrade, Serbia

    Nevena Pantić & Olivera Prodanović

  2. University of Belgrade, Innovative Centre of the Faculty of Chemistry, Studentski trg 12-16, 11000, Belgrade, Serbia

    Milica Spasojević

  3. Henkel Company, Bulevar Oslobođenja 383, 11040, Belgrade, Serbia

    Željko Stojanović

  4. University of Belgrade, Faculty of Technology and Metallurgy, Karnegijeva 4, 11000, Belgrade, Serbia

    Đorđe Veljović

  5. University of Belgrade, Institute of Chemistry, Technology and Metallurgy, Njegoševa 12, 11000, Belgrade, Serbia

    Jugoslav Krstić & Ana Marija Balaž

  6. University of Belgrade, Faculty of Chemistry, Studentski trg 12-16, 11000, Belgrade, Serbia

    Radivoje Prodanović

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  1. Nevena Pantić
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  2. Milica Spasojević
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Pantić, N., Spasojević, M., Stojanović, Ž. et al. Immobilization of Horseradish Peroxidase on Macroporous Glycidyl-Based Copolymers with Different Surface Characteristics for the Removal of Phenol. J Polym Environ 30, 3005–3020 (2022). https://doi.org/10.1007/s10924-021-02364-3

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