Purification and characterization of an extracellular chitosanase produced by Amycolatopsis sp. CsO-2 |
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| Institution: | 1. KU Leuven – University of Leuven, Department of Microbiology and Immunology, Laboratory of Molecular Bacteriology, Rega Institute for Medical Research, Herestraat 49, box 1037, B-3000 Leuven, Belgium;2. KU Leuven – University of Leuven, Department of Microbiology and Immunology, Laboratory of Clinical Bacteriology and Mycology, Herestraat 49, box 819, B-3000 Leuven, Belgium;3. KU Leuven – University of Leuven, Department of Microbiology and Immunology, Laboratory of Virology and Chemotherapy, Rega Institute for Medical Research, Herestraat 49, box 1043, B-3000 Leuven, Belgium;4. KU Leuven – University of Leuven, University Hospitals Leuven, Laboratory Medicine, Herestraat 49, box 7003, B-3000 Leuven, Belgium;1. School of Pharmaceutical Sciences and National Glycoengineering Research Center, Shandong University, Wenhuaxi Road 44, Jinan 250012, Shandong Province, PR China;2. School of Basic Medical Sciences, Shandong University, Wenhuaxi Road 44, Jinan 250012, Shandong Province, PR China;3. School of Pharmaceutical Analysis, Shandong University, Wenhuaxi Road 44, Jinan 250012, Shandong Province, PR China |
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| Abstract: | Extracellular chitosanase produced by Amycolatopsis sp. CsO-2 was purified to homogeneity by precipitation with ammonium sulfate followed by cation exchange chromatography. The molecular weight of the chitosanase was estimated to be about 27,000 using SDS-polyacrylamide gel electrophoresis and gel filtration. The maximum velocity of chitosan degradation by the enzyme was attained at 55°C when the pH was maintained at 5.3. The enzyme was stable over a temperature range of 0–50°C and a pH range of 4.5–6.0. About 50% of the initial activity remained after heating at 100°C for 10 min, indicating a thermostable nature of the enzyme. The isoelectric point of the enzyme was about 8.8. The enzyme degraded chitosan with a range of deacetylation degree from 70% to 100%, but not chitin or CM-cellulose. The most susceptible substrate was 100% deacetylated chitosan. The enzyme degraded glucosamine tetramer to dimer, and pentamer to dimer and trimer, but did not hydrolyze glucosamine dimer and trimer. |
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