Abstract
Enzyme immobilization is an operative method for improving enzyme properties. There are various supports to enzyme immobilization. In the present study chitosan nanoparticles have been applied to alkaline phosphatase (ALP) immobilization due to their special properties. Firstly, the ionic gelation method was used to synthesize chitosan nanoparticles, which were characterized using XRD and Zetasizer Nano. Then, chitosan nanoparticles were activated with 1-ethyl-3-(3-dimethylaminopropyl carbodiimide (EDC) and ALP was successfully immobilized on them. The influence of reaction time, pH, and concentration of enzyme and EDC on efficiency of immobilization has been investigated. The studied stability against organic solvents and high temperature indicated that the stability of immobilized enzyme was increased during immobilization. The surface topography of free nanoparticles and nanoparticles with immobilized enzyme was studied by AFM. The calculation of binding efficiency showed that the immobilized ALP has conserved 71% of its native activity. The improvement of enzyme stability by immobilization indicated that chitosan nanoparticles were suitable support for immobilization.
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