Recombinant expression of a chitosanase and its application in chitosan oligosaccharide production |
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Authors: | Ya-Li Liu Shu Jiang Hai-Shui Wu Zhan-Yun Guo |
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Affiliation: | a Institute of Protein Research, College of Life Sciences and Technology, Tongji University, 1239 Siping Road, Shanghai 200092, China b Zhejiang Feng’an Biopharmaceutical, Xianju 317300, China c Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China d Shanghai East Hospital, Tongji University, Shanghai 200120, China |
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Abstract: | Recently, considerable attention has been focused on chitosan oligosaccharides (COSs) due to their various biological activities. COSs can be prepared by enzymatic degradation of chitosan, which is the deacetylation product of chitin, one of the most abundant biopolymers in nature. In the current study, we recombinantly expressed a chitosanase and used it for COS preparation. A bacillus-derived GH8 family chitosanase with a 6×His tag fused at its N-terminal was expressed in the Escherichia coli strain BL21(DE3) as a soluble and active form. Its expression level could be as high as 500 mg/L. Enzymatic activity could reach approximately 140,000 U/L under our assay conditions. The recombinant chitosanase could be purified essentially to homogeneity by immobilized metal-ion affinity chromatography. The enzyme could efficiently convert chitosan into monomer-free COS: 1 g of enzyme could hydrolyze about 100 kg of chitosan. Our present work has provided a cheap chitosanase for large-scale COS production in industry. |
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Keywords: | Chitosanase Recombinant expression Chitosan oligosaccharides |
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