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Covalent immobilization of penicillin G acylase onto amine-functionalized PVC membranes for 6-APA production from penicillin hydrolysis process. II. Enzyme immobilization and characterization

Authors

  • M. S. Mohy Eldin,

    Corresponding author
    1. Polymer Materials Research Department, Advanced Technology and New Materials Research Institute (ATNMRI), City for Scientific Research and Technology Applications (CSRTA), New Burg Al Arab, 21934-Alexandria, Egypt
    • Polymer materials research Department, Advanced Technology and New Materials Research Institute (ATNMRI), City for Scientific Research and Technology Applications (CSRTA), New Burg Al Arab, 21934-Alexandria, Egypt
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  • H. A. El Enshasy,

    1. Bioprocess Development Department (BID), Genetic Engineering and Biotechnology Research Institute (GEBRI), City for Scientific Research and Technology Applications (CSRTA), New Burg Al Arab, 21934 Alexandria, Egypt
    2. Chemical Engineering Pilot Plant (CEPP), Faculty of Chemical Engineering and Natural Resources, University Technology Malaysia (UTM), 81310 Skudai, Johor, Malaysia
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  • M. E. Hassan,

    1. Polymer Materials Research Department, Advanced Technology and New Materials Research Institute (ATNMRI), City for Scientific Research and Technology Applications (CSRTA), New Burg Al Arab, 21934-Alexandria, Egypt
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  • B. Haroun,

    1. Chemistry Department, Faculty of Science, Al Azhar University, Cairo, Egypt
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  • E. A. Hassan

    1. Chemistry Department, Faculty of Science, Al Azhar University, Cairo, Egypt
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Abstract

This article describes the covalent immobilization of penicillin G acylase (PGA) onto glutaraldehyde-activated NH2-PVC membranes. The immobilized enzyme was used for 6-aminopenicillanic acid production from penicillin hydrolysis. Parameters affecting the immobilization process, which affecting the catalytic activity of the immobilized enzyme, such as enzyme concentration, immobilization's time and temperature were investigated. Enzyme concentration and immobilization's time were found of determine effect. Higher activity was obtained through performing enzyme immobilization at room temperature. Both optimum temperature (35°C) and pH (8.0) of immobilized enzyme have not been altered upon immobilization. However, immobilized enzyme acquires stability against changes in the substrate's pH and temperature values especially in the higher temperature region and lower pH region. The residual relative activities after incubation at 60°C were more than 75% compared to 45% for free enzyme and above 50% compared to 20% for free enzyme after incubation at pH 4.5. The apparent kinetic parameters KM and VM were determined. KM of the immobilized PGA (125.8 mM) was higher than that of the free enzyme (5.4 mM), indicating a lower substrate affinity of the immobilized PGA. Operational stability for immobilized PGA was monitored over 21 repeated cycles. The catalytic membranes were retained up to 40% of its initial activity after 10.5 working h. © 2012 Wiley Periodicals, Inc. J Appl Polym Sci, 2012

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