Bioconjugation as a smart immobilization approach for α-amylase enzyme using stimuli-responsive Eudragit-L100 polymer: a robust biocatalyst for applications in pharmaceutical industry
Enzymes are powerful versatile biocatalysts, however, industrial application of enzymes is usually hampered by their susceptibility. Bio-inspired Eudragit-α-amylase conjugate (E-AC) was proposed as a biocatalyst for various pharmaceutical and industrial applications. In this study, α -Amylase (E.C....
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Main Authors: | , , , , , , |
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Format: | Article |
Language: | English |
Published: |
Taylor & Francis Group
2019-12-01
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Series: | Artificial Cells, Nanomedicine, and Biotechnology |
Subjects: | |
Online Access: | https://www.tandfonline.com/doi/10.1080/21691401.2019.1626414 |
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Summary: | Enzymes are powerful versatile biocatalysts, however, industrial application of enzymes is usually hampered by their susceptibility. Bio-inspired Eudragit-α-amylase conjugate (E-AC) was proposed as a biocatalyst for various pharmaceutical and industrial applications. In this study, α -Amylase (E.C. 3.2.1.1) was immobilized by covalent conjugation to Eudragit L-100 under mild conditions. The effect of polymer, carbodiimide and enzyme concentrations on optimization of (E-AC) was investigated. In addition, characterization of the free α -Amylase and E-AC with regard to pH, temperature, kinetic parameters, reusability and operational and storage conditions was carried out. Results showed a shift of the optimum pH of E-AC towards the alkaline side whereas, E-AC exhibited higher thermal stability at all tested temperatures. The kinetic parameters, Km values were 2.87 mg/ml and 3.15 mg/ml and Vmax values were 8.35 mg/ml/min and 8.98 mg/ml/min for free and E-AC, respectively. E-AC retained 85% of the initial activity after five consecutive amylolytic cycles, thus emphasizing its powerful potentials. Operational storage and thermal stability were highly improved as well for E-AC conjugate with an 11.6 stabilization factor in comparison to the free α-amylase. In this study, Eudragit L-100 polymer was successfully used as smart immobilization support to create a reversibly soluble-insoluble enzyme biocatalyst to enforce and extend biotechnological applications of α-amylase in the pharmaceutical industry. |
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ISSN: | 2169-1401 2169-141X |