Immobilization and characterization of horseradish peroxidase into chitosan and chitosan/PEG nanoparticles: A comparative study |
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| Affiliation: | 1. Chemistry Department, Faculty of Sciences and Arts, University of Jeddah, Khulais, P.O. Box 355, Khulais 21921, Saudi Arabia;2. Chemistry Department, Faculty of Applied Science, Taiz University, Taiz, Yemen;3. Chemistry Department, Faculty of Science, King Abdulaziz University, Jeddah, P. O. Box 80200, Jeddah 21589, Saudi Arabia;4. Dyeing, Printing and Textile Auxiliaries Department, Textile Research Division, National Research Center, Dokki, 71516, Cairo, Egypt;5. Department of Biochemistry, Faculty of Science, King Abdulaziz University, Jeddah, P. O. Box 80200, Jeddah 21589, Saudi Arabia;6. Center of Nanotechnology, King Abdulaziz University, Jeddah, 21589, Saudi Arabia;7. Molecular Biology Department, National Research Centre, Cairo, Egypt;8. Department of Biochemistry, Faculty of Science, University of Jeddah, P.O. Box 80203, Jeddah 21589, Saudi Arabia;9. Biochemistry Department, Faculty of Veterinary Medicine, Zagazig University, Egypt;10. Department of Biological Sciences, Faculty of Science, King Abdulaziz University, P.O. Box 80203, Jeddah 21589, Saudi Arabia;1. School of Life Science and Food Engineering, Huaiyin Institute of Technology, Huaian, 223003, China;2. School of Chemistry and Chemical Engineering, State Key Laboratory of Metal Matrix Composites, Shanghai Jiao Tong University, Shanghai 200240, China;3. Tecnologico de Monterrey, School of Engineering and Sciences, Campus Monterrey, Ave. Eugenio Garza Sada 2501, Monterrey, N.L. CP 64849, Mexico;4. Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin University of Science and Technology, No 29, 13th, Avenue, Tianjin Economic and Technological Development Area (TEDA), Tianjin 300457, China |
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| Abstract: | Chitosan (CS) is considered a suitable biomaterial for enzyme immobilization. CS combination with polyethylene glycol (PEG) can improve the biocompatibility and the properties of the immobilized system. Thus, the present work investigated the effect of the PEG in the horseradish peroxidase (HRP) immobilization into chitosan nanoparticles from the morphological, physicochemical, and biochemical perspectives. CS and CS/PEG nanoparticles were obtained by ionotropic gelation and provided immobilization efficiencies (IE) of 65.8 % and 51.7 % and activity recovery (AR) of 76.4 % and 60.4 %, respectively. The particles were characterized by DLS, ZP, SEM, FTIR, TGA and DSC analysis. Chitosan nanoparticles showed size around 135 nm and increased to 229 nm after PEG addition and HRP immobilization. All particles showed positive surface charges (20−28 mV). Characterizations suggest nanoparticles formation and effective immobilization process. Similar values for optimum temperature and pH for immobilized HRP into both nanoparticles were found (45 °C, 7.0). Vmax value decreased by 5.07 to 3.82 and 4.11 mM/min and KM increased by 17.78 to 18.28 and 19.92 mM for free and immobilized HRP into chitosan and chitosan/PEG nanoparticles, respectively. Another biochemical parameters (Kcat, Ke, and Kα) evaluated showed a slight reduction for the immobilized enzyme in both nanoparticles compared to the free enzyme. |
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| Keywords: | Horseradish peroxidase Chitosan Nanoparticle Polyethylene glycol |
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