Immobilization of alcalase on polydopamine modified magnetic particles

Enzymes play a crucial role in medicine, industry, and agriculture. Alcalase, a protease, has found wide-ranging applications in both the detergent and food industries. Immobilizing enzymes has gained prominence as a technology to enhance enzyme stability and reusability, and magnetic particles (MP) have emerged as promising carriers for enzyme immobilization due to their magnetic properties and ease of synthesis. In our study, we propose a novel approach that utilizes polydopamine-modified magnetic particles (MPP) as carriers for immobilizing alcalase. The immobilization process entails modifying the magnetic particles with polydopamine and functionalizing them with glutaraldehyde (GA). We conducted experiments to determine the optimal conditions for alcalase immobilization. These conditions were identified as a pH of 7.5, a GA concentration of 0.23 μg/mL, an alcalase concentration of 6.1 mg/mL, and an immobilization time of 4 hours. The immobilized alcalase significantly improved its temperature and pH stability. Furthermore, kinetic studies of the immobilized enzyme were conducted, revealing that while the Michaelis constant (Km) remained unaffected, there was a decrease in the maximum velocity (Vmax). After 14 repeated uses, it retained 78.66% of its relative activity. This innovative strategy not only enhances our understanding of enzyme immobilization techniques but also offers new avenues for leveraging enzymes in a multitude of applications.