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Immobilization of Horseradish Peroxidase on Modified Cellulose Carriers via Hydrophobic Interactions: Catalytic Properties and Stability

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Abstract

Cellulose is one of the most common biopolymers on earth with well-known non-toxic, biocompatible, and environmentally friendly properties. Structural and chemical properties of cellulose fibers give additional opportunities for surface modification of cellulose and its use as a carrier for immobilization of various bioactive compounds, including enzymes. For the proposes of horseradish peroxidase (HRP) immobilization, five different cellulose-based carriers were synthesized by acylation of microcrystalline cellulose. After HRP immobilization, based on the saved catalytic activity, protein equilibrium, and immobilization parameters, the benzoyl- and cinnamoyl-cellulose carriers were selected for further characterization. Kinetic studies have shown that the apparent Michaelis constants for HRP immobilized on benzoyl-cellulose and cinnamoyl-cellulose carriers were 58.1 µM and 60.8 µM for hydrogen peroxide and 113.4 µM and 115.2 µM for ABTS, respectively. Experimental investigations of temperature and pH value influence on immobilized HRP on two selected cellulose carriers have shown that the optimal values of temperature and pH were 35 °C and 6.8, respectively. Thermal and storage stability tests showed that immobilized HRP could exhibit improved thermal and storage stability compared to the free enzyme. Operational stability tests showed that HRP immobilized on modified cellulose carriers can do up to 20 successive batch operations, without the decrease in initial activity during the first eight cycles.

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Acknowledgements

This work was supported by Republic of Serbia—Ministry of Education, Science and Technological Development, Program for Financing Scientific Research Work, ev. No. 451-03-68/2020-14/200133.

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Authors and Affiliations

  1. Faculty of Technology, University of Nis, Bulevar Oslobodjenja 124, 16000, Leskovac, Serbia

    Sasa Savic, Sanja Petrovic & Zivomir Petronijevic

  2. DCP Hemigal, Tekstilna 97, 1600, Leskovac, Serbia

    Sanela Savic

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  1. Sasa Savic
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  2. Sanja Petrovic
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  3. Sanela Savic
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  4. Zivomir Petronijevic
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Correspondence to Sasa Savic.

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Savic, S., Petrovic, S., Savic, S. et al. Immobilization of Horseradish Peroxidase on Modified Cellulose Carriers via Hydrophobic Interactions: Catalytic Properties and Stability. Iran J Sci Technol Trans Sci 45, 55–63 (2021). https://doi.org/10.1007/s40995-020-01027-7

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  • DOI: https://doi.org/10.1007/s40995-020-01027-7

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