Immobilization of Laccase from Trametes hirsuta onto CMC Coated Magnetic Nanoparticles

Abstract

In this study, Fe3O4/CMC magnetic nanoparticles were synthesized through co-precipitation method. Afterward, laccase from Trametes hirsuta was immobilized onto Carboxymethyl cellulose (CMC)coated magnetic Fe3O4 nanoparticles by covalent bonding between carboxyl groups of carboxymethyl cellulose and amine group of laccases. Also, the resulted magnetic nanoparticles and immobilized laccase were characterized by Fourier-transform infrared spectroscopy (FTIR), scanning electron microscope (SEM) and dynamic light scattering (DLS) analysis. Moreover, the vital factors in enzyme immobilization, such as contact time, amount of N-hydroxysuccinimide (NHS), and the amount of nanoparticles were optimized, which successively 48 h, 0.01 g, and 0.0125 g were achieved for 0.01g of N-ethyl-N′-(3-dimethylaminopropyl)carbodiimide (EDC). Activity recovery of 51 ± 0.8% was achieved by optimizing the immobilization process. The results also indicated that the loading of laccase onto carboxymethyl cellulose-coated Fe3O4 nanoparticles was approximately 120 (mg/g). Finally, the immobilized laccases on magnetic support could save nearly 50% of their initial activity after five consecutive cycles.

Enhancing Enzymatic Activity Through Co-Immobilization of Protease and Lipase Enzymes on Magnetic Nano-Iron Oxides Coated with Gum Arabic

“…Various techniques, such as adsorption, entrapment, covalent bonding, and cross-linking, have been used to immobilize enzymes on magnetic nanoparticles, each offering unique advantages in terms of IOP Publishing doi:10.1088/1755-1315/1371/6/062046 2 enzyme activity and stability [20,23,29].In addition, the use of immobilization methods based on coated magnetic nanoparticles has shown promising results in achieving high enzyme activity and stability [21]. The application of magnetic nanoparticles to stabilize multiple enzymes has been demonstrated in the development of multienzyme biocatalysts, providing synergistic effects and improving the analytical performance of these enzymes [17,30].The use of magnetic nanoparticles has also enabled the development of magnetically separable enzyme catalyst systems, allowing easy recovery and reuse of immobilized enzymes [25,26,27,28]. Given that the immobilized enzyme has many economic, environmental and technical advantages, including operational stability and reusability, this study aimed to co-immobilize the enzymes protease and lipase simultaneously using magnetic nano-iron oxides to increase the activity of the enzymes and to achieve the largest diffusion distance for the enzymes using ultrasound in addition to the ability to easily withdraw enzymes from the reaction medium at any moment using external magnetic flux.…”