Immobilization of acetylcholinesterase on new modified acrylonitrile copolymer membranes |
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| Institution: | 1. University “Prof. Dr. A. Zlatarov”, Department of Biotechnology, Prof Y. Yakimov Str. 1, 8010 Bourgas, Bulgaria;2. Laboratory of Bioactive Polymers, Institute of Polymers, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria;1. China National Rice Research Institute, Hangzhou 310006, China;2. Institute of Microanalytical Systems, Department of Chemistry, Zhejiang University, Zijin’gang Campus, Hangzhou 310058, China;3. Laboratory of Quality & Safety Risk Assessment for Rice (Hangzhou), Ministry of Agriculture, Hangzhou, China;1. Key Laboratory of Luminescent and Real-Time Analytical Chemistry, Ministry of Education (Southwest University), College of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715, PR China;2. Institute for Clean Energy & Advanced Materials, Faculty of Materials and Energy, Southwest University, Chongqing 400715, PR China;1. Beijing National Laboratory for Molecular Sciences; Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences, No. 2 Zhongguancun Beiyijie, Beijing 100190, PR China;2. Department of Chemistry, Yonsei University, 50 Yonsei-ro, Seoul 120-749, South Korea |
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| Abstract: | The three new dual-layer matrices (polyacrylonitrile (PAN) membranes coated with physically bound chitosan (CHI)—PANCHI-A and chemically bound chitosan—PANCHI-B and PANCHI-C) for immobilization of acetylcholinesterase (AChE) were obtained. The chemical-modified PAN membrane (PAN-NaOH + ethylenediamine (EDA)) was used as a base for the prepared dual-layer membranes. For chemical chitosan bound membrane, chitosan was tethered onto the membrane surface to form a dual-layer biomimetic membrane in the presence of glutaraldehyde (GA). The basic characteristics (amount of amino groups, hydrophilicity and transport characteristics) of the chitosan-modified membranes were investigated. The SEM analyses were shown essential morphology change in the different chitosan membranes.The relative activities and Vmax of the covalently immobilized enzyme on PANCHI-B and PANCHI-C membranes were higher than that on PANCHI-A membrane and chemical-modified membrane with NaOH + EDA. Km values for the different modified membranes are lower for the chitosan-treated membranes. The pH and temperature optimum of immobilized enzyme were determined. The bound enzymes on PANCHI-B and PANCHI-C have higher thermal and storage stability in comparison with AChE on PANCHI-A membrane and free enzyme. |
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