Abstract
Abstract Magnetic chitosan/Fe 3 O 4 nanoparticles prepared by in situ mineralization of Fe 3 O 4 in chitosan hydrogel were first modified by 3-aminopropyltriethoxysilane (3-APTES) and then tested for the possibility of being used as carriers for the immobilization of pullulanase. It was found that the immobilized enzyme showed high activity retention (>70%) and the affinity between enzyme and substrate was improved. However, compared to the enzyme immobilized by electrostatic adsorption (87% of its initial activity lost after 10 consecutive reuses), the enzyme immobilized by crosslinking was more stable and there was less loss of activity. The improvement of activity retention and stability was further studied from the view of surface amino concentration of nanoparticles at the function of the amount of 3-APTES added, and crosslinker concentration in the activation process. The highest activity retention (89%) and operation stability (61% residual activity after 10 consecutive reuses) were obtained under the condition of 1.6 amino groups per nm 2 and 15% glutaraldehyde (v/v, dimmer of glutaraldehyde). Therefore, the magnetic chitosan/Fe 3 O 4 nanoparticle prepared by in situ mineralization was proved to be a suitable carrier. In addition, immobilized pullulanase exhibited good resistance to metal ions and detergents.
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