Journal
COLLOIDS AND SURFACES B-BIOINTERFACES
Volume 202, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.colsurfb.2021.111692
Keywords
Chitosanase; Crystal structure; Rational design; Chitobiose
Funding
- Science and Technology Department of Sichuan Province [2020ZYD024]
- Innovative Academy of Seed Design (INASEED) at the Chinese Academy of Sciences
- 100 Talents Program of the Chinese Academy of Sciences
- Key Laboratory of Environmental and Applied Microbiology at the Chinese Academy of Sciences
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This research successfully prepared pure chitosan oligosaccharides using chitosanase from Bacillus subtilis MY002, providing a foundation for its future biomedical applications.
Chitosan oligosaccharides (COS) are attractive active molecules for biomedical applications. Currently, the prohibitively high cost of producing fully defined COS hampers extensive studies on their biological activity and restricts their use in various industries. Thus, cost-effective production of pure COS is of major importance. In this report, chitosanase from Bacillus subtilis MY002 (CsnMY002) was prepared for COS production. The structure of apo CsnMY002 displayed an unexpected tunnel-like substrate-binding site and the structure of the CsnMY002_E19A/(GlcN)6 complex highlighted the 4 + 2 '' splitting of hexaglucosamine even though the 3 + 3 '' splitting is also observed in the TLC analysis of the enzyme products for hexaglucosamine. Structure based rational design was performed to generate mutants for chitobiose production. The CsnMY002_G21 K mutant produced chitobiose with a relative content > 87 % from chitosan with a low degree of acetylation, and 50.65 mg chitobiose with a purity > 98 % was prepared from 100 mg chitosan. The results provide insight on the catalytic mechanism of chitosanase and underpin future biomedical applications of pure chitobiose.
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