Efficient digestion of chitosan using chitosanase immobilized on silica-gel for the production of multisize chitooligosaccharides |
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| Institution: | 1. Department of Pharmaceutical Science, Gachon University, Incheon 406-799, Republic of Korea;2. Department of Biological Sciences, Ohio University, Zanesville, OH 43701, USA;3. Diabetes Institute, Ohio University, Athens, OH 45701, USA;4. Department of Life Science, College of Bionano, Gachon University, Incheon 406-799, Republic of Korea;1. Department of Environmental and Food Sciences, Fukui University of Technology, Fukui 910-8505, Japan;2. Institue for Protein Research, Osaka University, Suita 565-0871, Japan;3. Department of Advanced Bioscience, Kinki University, Nara 631-8505, Japan;4. Department of Bioscience, Fukui Prefectural University, Eiheiji-cho, Yoshida-gun, Fukui 910-1195, Japan;1. Department of Chemical and Environmental Engineering, University of Toledo, Toledo, OH 43606, United States;2. School of Green Chemistry and Engineering, University of Toledo, Toledo, OH 43606, United States |
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| Abstract: | Chitosanase-coated silica-gels were prepared via cross-linking of the chitosanase onto silica-gels for the efficient production of multisize chitooligosaccharides (MCOs) in a continuous process. The kinetic aspects of immobilized chitosanase (IMMCTase) were investigated based on the reaction time, production of MCOs, and MALDI-TOF mass analyses to achieve maximum bioconversion of high molecular weight chitosan (HMWC) to MCOs. IMMCTase revealed a negligible loss of chitosanase activity after multi uses in continuous digestion of HMWC. The optimal temperature of IMMCTase was 37 °C, and kinetic parameters toward HMWC were determined to be Km = 1.45 mM and Vmax = 360 μmole/μg/min, respectively. Under optimal conditions, the recovery of enzyme activity of IMMCTase was determined to be 82.3%, thus indicating that it can still be reused few more times. In conclusion, use of IMMCTase resulted in rapid and efficient digestions of HMWC with consistent results to produce MCOs. |
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| Keywords: | Multisize chitooligosaccharides Chitosanase Immobilized enzyme MALDI-TOF Kinetic parameters |
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