Preparation and characterization of Fe₃O₄ nanoparticles via green synthesis using firethorn (pyracantha) fruit extract and investigation of their usability in α-amylase enzyme immobilization.

The production of metal oxide (Fe₃O₄) compounds via the green synthesis method offers significant advantages over other techniques due to its environmentally friendly, sustainable, practical, economical, and safe nature. Nanoparticles synthesized by green methods tend to exhibit enhanced biocompatibility, making them suitable for medical and biomedical applications. In this study, the primary objective is to obtain Fe₃O₄ nanoparticles through green synthesis using an aqueous extract of Pyracantha (firethorn) fruit. Following the green synthesis of Fe₃O₄ nanoparticles, the immobilization of the enzyme α-amylase onto these nanoparticles is targeted. Immobilized α-amylase has broad applications in various industrial and laboratory contexts, specifically for starch hydrolysis. This immobilized enzyme is advantageous as it is reusable, exhibits high stability, and enhances efficiency in production processes.Due to its starch-hydrolyzing effect, immobilized α-amylase can be utilized in sectors such as sweetener production, the paper industry, and pharmaceuticals. The enzyme will be covalently immobilized, and its optimum pH and temperature, along with stability tests (pH, temperature, and storage stability), operational stability, and reusability, will be examined. Furthermore, enzymatic kinetic parameters in starch hydrolysis will be determined. Structural analysis (characterization) of the Fe₃O₄ nanoparticles will be conducted using FTIR, SEM, TGA, zeta potential, and XRD techniques.