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Immobilization of Alkaline Phosphatase Using Chitosan Nanoparticles for Enhancing Its Stability Publisher



Jafary F1 ; Varshosaz J2 ; Panjehpour M3 ; Yaghmaei P1
Authors
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Authors Affiliations
  1. 1. Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran
  2. 2. Department of Pharmaceutics, School of Pharmacy and Novel Drug Delivery Systems Research Centre, Isfahan University of Medical Sciences, Isfahan, Iran
  3. 3. Department of Clinical Biochemistry, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran

Source: Russian Journal of Applied Chemistry Published:2014


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. © 2014 Pleiades Publishing, Ltd.
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