Abstract
Cascade reactions catalyzed by multi-enzymatic systems have attracted enormous scientific interest over the last decade. They are an emerging technology that significantly expands the applicability of biocatalysts in several biotechnological processes, such as the synthesis of high value-added products. Immobilization of enzymes on a solid carrier is a commonly used strategy to improve the stability and reuse of multiple enzyme systems. Magnetic nanoparticles have been applied as promising nanocarriers for either the immobilization of one enzyme or the co-immobilization of multiple enzymes. In this chapter, we describe the preparation of magnetic iron oxide nanoparticles γ-Fe2O3 modified with 3-(aminopropyl)-triethoxysilane (APTES), for the simultaneous covalent co-immobilization of oxidoreductases and hydrolytic enzymes, such as cellulase, β-glucosidase (bgl), glucose oxidase (GOx), and horseradish peroxidase (HRP). Several spectroscopic techniques that are used to characterize the structure and the catalytic performance of such systems are also described.
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Acknowledgements
This work was supported by the project “Synthetic Biology: from omics technologies to genomic engineering (OMIC-ENGINE)” (MIS 5002636), which is implemented under the Action “Reinforcement of the Research and Innovation Infrastructure,” funded by the Operational Program “Competitiveness, Entrepreneurship and Innovation” (NSRF 2014–2020) and co-financed by Greece and the European Union (European Regional Development Fund).
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Giannakopoulou, A., Patila, M., Chalmpes, N., Polydera, A.C., Gournis, D., Stamatis, H. (2022). A Four-enzyme Nanoassembly Consisting of Hydrolases and Oxidoreductases for Multi-step Cascade Reactions. In: Stamatis, H. (eds) Multienzymatic Assemblies. Methods in Molecular Biology, vol 2487. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2269-8_16
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DOI: https://doi.org/10.1007/978-1-0716-2269-8_16
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