Effects of Incubation Time and Laccase Concentration on Immobilization of Laccase on Magnetic Spent Tea
DOI:
https://doi.org/10.11113/bioprocessing.v3n2.64Keywords:
magnetic spent tea, laccase, immobilizationAbstract
This study explores the impact of incubation time and laccase concentration on the immobilization efficiency and activity of laccase, employing magnetic spent tea as a sustainable carrier. The results indicate that the highest immobilization yield occurred at an incubation time of 24 hours, achieving 96.51%, while the optimal enzyme activity of 4.31 U was recorded at a shorter incubation duration of 6 hours. This finding suggests that extended incubation times may enhance covalent bonding between the enzyme and the carrier but can also lead to reduced enzymatic activity due to potential over-binding. The ideal laccase concentration was identified as 1.0 mg/ml, which resulted in a notable immobilization yield of 74.16% while preserving significant enzyme activity. Higher concentrations caused steric hindrance, adversely affecting performance. Furthermore, reusability assessments revealed that the immobilized laccase's relative activity increased from 23% in the first cycle to 100% by the fifth cycle, likely facilitated by ABTS as a mediator. Utilizing magnetic spent tea not only serves as an effective and economical method for enzyme immobilization but also aids in waste reduction by transforming tea byproducts into valuable resources. This dual approach underscores the potential for enhancing enzymatic processes while advancing environmentally responsible waste management practices in biotechnological applications.
References
Al-sareji, O. J., Meiczinger, M., Al-Juboori, R. A., Grmasha, R. A., Andredaki, M., Somogyi, V., Idowu, I. A., Stenger-Kovács, C., Jakab, M., Lengyel, E., and Hashim, K. S. 2023a. Efficient Removal of Pharmaceutical Contaminants from Water and Wastewater using Immobilized Laccase on Activated Carbon Derived from Pomegranate Peels. Scientific Reports, 13(1). https://doi.org/10.1038/s41598-023-38821-3
Al-sareji, O. J., Meiczinger, M., Somogyi, V., Al-Juboori, R. A., Grmasha, R. A., Stenger-Kovács, C., Jakab, M., and Hashim, K. S. 2023b. Removal of Emerging Pollutants from Water using Enzyme-Immobilized Activated Carbon from Coconut Shell. Journal of Environmental Chemical Engineering, 11(3). https://doi.org/10.1016/j.jece.2023.109803
Arica, M. Y., Salih, B., Celikbicak, O., and Bayramoglu, G. (2017). Immobilization of Laccase on the Fibrous Polymer-Grafted Film and Study of Textile Dye Degradation by MALDI–ToF-MS. Chemical Engineering Research and Design, 128, 107–119. https://doi.org/10.1016/j.cherd.2017.09.023
Ba, S., Arsenault, A., Hassani, T., Jones, J. P., & Cabana, H. 2013. Laccase Immobilization and Insolubilization: from Fundamentals to Applications for the Elimination of Emerging Contaminants in Wastewater Treatment. Critical Reviews in Biotechnology, 33(4), 404–418. https://doi.org/10.3109/07388551.2012.725390
Bayramoglu, G., Yilmaz, M., and Yakup Arica, M. (2010). Preparation and Characterization of Epoxy-Functionalized Magnetic Chitosan Beads: Laccase Immobilized for Degradation of Reactive Dyes. Bioprocess and Biosystems Engineering, 33(4), 439–448. https://doi.org/10.1007/s00449-009-0345-6
Cañas, A. I., and Camarero, S. 2010. Laccases and Their Natural Mediators: Biotechnological Tools for Sustainable Eco-Friendly Processes. Biotechnology Advances, 28(6), 694–705. https://doi.org/10.1016/j.biotechadv.2010.05.002
Deng, J., Wang, H., Zhan, H., Wu, C., Huang, Y., Yang, B., Mosa, A., and Ling, W. 2022. Catalyzed Degradation of Polycyclic Aromatic Hydrocarbons by Recoverable Magnetic Chitosan Immobilized Laccase from Trametes Versicolor. Chemosphere, 301. https://doi.org/10.1016/j.chemosphere.2022.134753
Franssen, M. C. R., Steunenberg, P., Scott, E. L., Zuilhof, H., and Sanders, J. P. M. 2013. Immobilised Enzymes in Biorenewables Production. Chemical Society Reviews, 42(15), 6491. https://doi.org/10.1039/c3cs00004d
Gao, Y., Shah, K., Kwok, I., Wang, M., Rome, L. H., and Mahendra, S. 2022. Immobilized Fungal Enzymes: Innovations and Potential Applications in Biodegradation and Biosynthesis. Biotechnology Advances, 57, 107936. https://doi.org/10.1016/j.biotechadv.2022.107936
Girelli, A. M., and Scuto, F. R. 2021a. Spent Grain as a Sustainable and Low-Cost Carrier for Laccase Immobilization. Waste Management, 128, 114–121. https://doi.org/10.1016/j.wasman.2021.04.055
Girelli, A. M., and Scuto, F. R. (2021b). Spent Grain as a Sustainable and Low-Cost Carrier for Laccase Immobilization. Waste Management, 128, 114–121. https://doi.org/10.1016/j.wasman.2021.04.055
Gu, Y., Yuan, L., Jia, L., Xue, P., and Yao, H. 2021. Recent Developments of a Co-Immobilized Laccase-Mediator System: A Review. In RSC Advances (Vol. 11, Issue 47, pp. 29498–29506). Royal Society of Chemistry. https://doi.org/10.1039/d1ra05104k
Hameed, B. H. 2009. Spent Tea Leaves: A New Non-Conventional and Low-Cost Adsorbent for Removal of Basic Dye from Aqueous Solutions. Journal of Hazardous Materials, 161(2–3), 753–759. https://doi.org/10.1016/j.jhazmat.2008.04.019
Homaei, A. A., Sariri, R., Vianello, F., and Stevanato, R. (2013). Enzyme Immobilization: An Update. Journal of Chemical Biology, 6(4), 185–205. https://doi.org/10.1007/s12154-013-0102-9
Humphrey, S., & Yakubu, S. (2024). A Review on Tea Leaves and Tea Fibre as Adsorbents for Heavy Metals Removal. Journal of Scientific Insights, 1(2), 62-78.
Imam, A., Suman, S. K., Singh, R., Vempatapu, B. P., Ray, A., & Kanaujia, P. K. (2021). Application of Laccase Immobilized Rice Straw Biochar for Anthracene Degradation. Environmental Pollution, 268, 115827.
Li, N., Xia, Q., Niu, M., Ping, Q., and Xiao, H. 2018. Immobilizing Laccase on Different Species Wood Biochar to Remove the Chlorinated Biphenyl in Wastewater. Scientific Reports, 8(1). https://doi.org/10.1038/s41598-018-32013-0
Mohd Syukri, M. S., Rahman, R. A., Mohamad, Z., Nik Mahmood, N. A., Illias, R. M., and Tokuyama, H. 2020. Laccase Immobilisation on Poly(Ethylene) Terephthalate Grafted with Maleic Anhydride (PET-G-MAH) Nanofiber Mat. Chemical Engineering Transactions, 78, 37–42. https://doi.org/10.3303/CET2078007
Mozaffari, M., Khodabakhshi, A., and Azizi, A. 2023. Evaluation of Pb (II) Removal by Tea Pulp Modified with Magnetite Nanoparticle. Journal of Chemistry,. https://doi.org/10.1155/2023/7011995
Oraby, K. R. M., Villalonga, A., Hassan, F. S. M., Zayed, M. A., Mubarak, M. F., Ojeda, I., Sánchez, A., and Villalonga, R. 2025. Immobilization of Laccase on Fe3O4@SiO2 core@shell Magnetic Nanoparticles for Methylene Blue Biodegradation. Process Biochemistry, 148, 10–16. https://doi.org/10.1016/j.procbio.2024.11.012
Panneerselvam, P., Morad, N., and Tan, K. A. 2011. Magnetic Nanoparticle (Fe3O4) impregnated onto Tea Waste for the Removal of Nickel(II) from Aqueous Solution. Journal of Hazardous Materials, 186(1), 160–168. https://doi.org/10.1016/j.jhazmat.2010.10.102
Syukri, M. S. M., Rahman, R. A., Latif, W. M. S. M., Mohamad, N., Abdullah, N., Bolong, N., Sulaiman, M. F., and Ibrahim, M. A. 2023. Implementation of Electrospun Nanofiber Mat as Enzyme Immobilization Carrier. AIP Conference Proceedings, 2945(1). https://doi.org/10.1063/5.0182085
Yaashikaa, P. R., Senthil Kumar, P., and Karishma, S. 2022. Review on Biopolymers and Composites – Evolving Material as Adsorbents in Removal of Environmental Pollutants. Environmental Research, 212. https://doi.org/10.1016/j.envres.2022.113114
Yang, A. L., Yang, S. Y., and Zhu, Y. K. 2021. Magnetic Modification of Used Tea Leaves for Uranium Adsorption. Xinxing Tan Cailiao/New Carbon Materials, 36(4), 821–826. https://doi.org/10.1016/S1872-5805(21)60053-7
Zhang, Y., Wang, Q., and Hess, H. 2017. Increasing Enzyme Cascade Throughput by pH-Engineering the Microenvironment of Individual Enzymes. ACS Catalysis, 7(3), 2047–2051. https://doi.org/10.1021/acscatal.6b03431