Covalent immobilization of acetylcholinesterase on a novel polyacrylic acid‐based nanofiber membrane
Autor: | Aslı Beyler Çiğil, Bekir Çakıroğlu, M. Vezir Kahraman, Ayşe Ogan, Serap Demir |
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Přispěvatelé: | Cakiroglu, Bekir, Cigil, Asli Beyler, Ogan, Ayse, Kahraman, M. Vezir, Demir, Serap |
Jazyk: | angličtina |
Rok vydání: | 2018 |
Předmět: |
GLUCOSE-OXIDASE
Environmental Engineering Immobilized enzyme SURFACE NF membrane Synthetic membrane Bioengineering 02 engineering and technology 010402 general chemistry 01 natural sciences Covalent immobilization chemistry.chemical_compound BIOMEDICAL APPLICATIONS Polyacrylic acid Research Articles MAGNETIC NANOPARTICLES biology Electrospinning STABILITY ELECTRODE POLY(ACRYLIC ACID) PERFORMANCE 021001 nanoscience & nanotechnology Acetylcholinesterase HYDROXYAPATITE Enzyme assay 0104 chemical sciences Membrane chemistry Nanofiber biology.protein 0210 nano-technology Carbonyldiimidazole Biotechnology Nuclear chemistry |
Zdroj: | Eng Life Sci |
Popis: | In this study, polyacrylic acid‐based nanofiber (NF) membrane was prepared via electrospinning method. Acetylcholinesterase (AChE) from Electrophorus electricus was covalently immobilized onto polyacrylic acid‐based NF membrane by demonstrating efficient enzyme immobilization, and immobilization capacity of polymer membranes was found to be 0.4 mg/g. The novel NF membrane was synthesized via thermally activated surface reconstruction, and activation with carbonyldiimidazole upon electrospinning. The morphology of the polyacrylic acid‐based membrane was investigated by scanning electron microscopy, Fourier Transform Infrared Spectroscopy, and thermogravimetric analysis. The effect of temperature and pH on enzyme activity was investigated and maxima activities for free and immobilized enzyme were observed at 30 and 35°C, and pH 7.4 and 8.0, respectively. The effect of 1 mM Mn(2+), Ni(2+), Cu(2+), Zn(2+), Mg(2+), Ca(2+) ions on the stability of the immobilized AChE was also investigated. According to the Michaelis–Menten plot, AChE possessed a lower affinity to acetylthiocholine iodide after immobilization, and the Michaelis–Menten constant of immobilized and free AChE were found to be 0.5008 and 0.4733 mM, respectively. The immobilized AChE demonstrated satisfactory reusability, and even after 10 consecutive activity assay runs, AChE maintained ca. 87% of its initial activity. Free enzyme lost its activity completely within 60 days, while the immobilized enzyme retained approximately 70% of the initial activity under the same storage time. The favorable reusability of immobilized AChE enables the support to be employable to develop the AChE‐based biosensors. |
Databáze: | OpenAIRE |
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