Preparation and Application of Carboxyl-Functioned Magnetic Nanoparticles for Arylsulfatase Immobilization
Arylsulfatase, one of a few enzymes that can enhance the gelling strength of agar by cleaving the sulfate ester bonds in agar, was covalently immobilized with carboxyl functioned magnetic nanoparticles (CMNPs). The resultant CMNPs and immobilized arylsulfatase were characterized by transmission electron microscopy (TEM), Dynamic Light scattering (DLS), X-ray diffraction (XRD), vibrating sample magnetometry (VSM) and thermogravimetric analysis (TGA). The TEM result indicated that the CMNPs and immobilized arylsulfatase had a similar mean particle size of 10 nm. The arylsulfatase-CMNPs had a mean diameter of 1200 nm in aqueous solution determined by the DLS, which was much bigger than the CMNPs (433.6 nm). The different sizes demonstrated that the arylsulfatase was coated on CMNPs successfully. XRD showed that diffraction patterns of the CMNPs and arylsulfatase-CMNPs were close to the standard XRD pattern of Fe3O4. Saturation magnetizations were 52.1 emu/g for carriers and 47.9 emu/g for immobilized arylsulfatase, which indicated that the particles had superparamagnetic characteristics. The TGA revealed that the amount of arylsulfatase bound to the surface of CMNPs was 5.65%. The arylsulfatase exhibited better thermal stability and reusability after immobilization, the immobilized arylsulfatase can retain more than 50% enzyme activity up to the 9th cycle.