Immobilization of cyclodextrin glucanotransferase on capillary membrane |
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| Affiliation: | 1. Enzymology and Fungal Biotechnology Lab, Botany and Microbiology Department, Faculty of Science, Zagazig University, Zagazig 44519, Egypt;2. Applied Nucleic Acids Research Center, Chemistry Department, Faculty of Science, Zagazig University, Egypt;3. Biochemistry Department, Faculty of Agriculture, Zagazig University, Egypt;1. Department of Biochemistry and Center for Excellence in Protein and Enzyme Technology, Faculty of Science, Mahidol University, Bangkok, Thailand;2. School of Biomolecular Science and Engineering, Vidyasirimedhi Institute of Science and Technology (VISTEC), Rayong, Thailand;3. Department of Chemistry, Faculty of Science, Chiang Mai University, Chiang Mai, Thailand;4. Institute for Innovative Learning, Mahidol University, Nakhon Pathom, Thailand;5. Biotechnology Research Center and Department of Biotechnology, Toyama Prefectural University, Imizu, Japan;6. Department of Biotechnology, College of Life Sciences, Ritsumeikan University, Japan;1. Department of Biochemistry, Chulalongkorn University, Patumwan, Bangkok, Thailand;2. Skeletal Disorders Research Unit, Faculty of Dentistry, Chulalongkorn University, Patumwan, Bangkok, Thailand;3. Laboratorium für Mikrobiologie, Fachbereich Biologie and Synmikro, Philipps-Universität, Marburg, Germany;4. Max-Plank-Institut für terrestrische Mikrobiologie, Marburg, Germany;5. School of Biomolecular Science and Engineering, Vidyasirimedhi Institute of Science and Technology (VISTEC), Rayong, Thailand;3. From the Departments of Biochemistry and;6. the Structural Biology Core, University of Missouri, Columbia, Missouri 65211,;4. the Department of Biochemistry, Virginia Tech, Blacksburg, Virginia 24061, and;5. the Department of Chemistry, Faculty of Science, Damietta University, Damietta 34517, Egypt |
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| Abstract: | A bioreactor system with the enzyme immobilized on a capillary membrane is a promising tool for the mass production of valuable substances, because of the good productive efficiency. To investigate the kinetics of immobilized cyclodextrin glucanotransferase ([EC 2.4.1.19]; CGTase) on a capillary membrane in a bioreactor system, the amount of immobilized CGTase and the operating conditions, such as pressure and the reaction temperature, were examined under a constant substrate concentration (1.0%) and a constant flow rate (0.12 m/s). When the CGTase was immobilized at a concentration of 0.04 to 0.62 mg per membrane area (cm2), the decrease in the immobilized amount of CGTase resulted in an increase in the cyclodextrin production rate (g of CD/h·m2; CPR) and the CPR correlated well with the flux of the CGTase-immobilized membrane. Although a higher reaction temperature caused an increase in the CPR within a short operating time of the bioreactor, repeated operation at 60°C led to a reduction in the CPR due to the denaturation of the immobilized CGTase. The percentage of cyclodextrin (CD) to total sugar obtained in the permeate was slightly more than 60% under most operating conditions, but immobilization of the excess amount of CGTase (0.42–0.62 mg/cm2) reduced the CD yield as well as the ratio of α-CD to β-CD, suggesting that it led to a CGTase side-reaction such as intermolecular transglycosylation. These data suggest that the conditions under which the bioreactor with 0.04–0.40 mg/cm2 was operated; a reaction temperature of 50°C, a residence time of 1–2 min and adjustable pressure, could be employed to obtain a high CPR using a large scale CGTase-immobilized membrane bioreactor. |
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