Biochemical characterization and cleavage pattern analysis of a novel chitosanase with cellulase activity

• Published in: Applied microbiology and biotechnology Vol. 106; no. 5-6; pp. 1979 - 1990
• Main Authors: Su, Haipeng, Sun, Jianan, Chu, Wenqin, Yuan, Bing, Mao, Xiangzhao
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.03.2022; Springer; Springer Nature B.V
• Subjects: Analysis; Biomedical and Life Sciences; Biotechnologically Relevant Enzymes and Proteins; Biotechnology; Cellulase; Chitosan; Chitosanase; Cleavage; Composition; E coli; Enzymes; Escherichia coli; Gene expression; Genetic aspects; Glycosidases; Glycoside hydrolase; Hydrolase; Hydrolases; Identification and classification; Life Sciences; Microbial Genetics and Genomics; Microbiology; Molecular docking; Oligosaccharides; Pattern analysis; Properties; Scission (Chemistry); Structure; Substrates; Sugar; China; Transglycosylation; GH5; Cellulase; Cleavage pattern; Chitosanase; Subsites
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-022-11829-x

Chitosanases are critical tools for the preparation of active oligosaccharides, whose composition is related to the cleavage pattern of the enzyme. Although numerous chitosanases have been characterized, the glycoside hydrolase (GH) family 5 chitosanases with other activities have rarely been investigated. Herein, a novel and second GH5 chitosanase OUC-Csngly from Streptomyces bacillaris was cloned and further characterized by expression in Escherichia coli BL21 (DE3). Interestingly, OUC-Csngly possessed dual chitosanase and cellulase activities. Molecular docking analysis showed that the C-2 group of sugar units affected the binding of the enzyme to oligosaccharides, which could result in different cleavage patterns toward chito-oligosaccharides (COSs) and cello-oligosaccharides. Further, we characterized OUC-Csngly’s distinctive cleavage patterns toward two different types of oligosaccharides. Meanwhile, endo-type chitosanase OUC-Csngly generated (GlcN) − (GlcN) 4 from chitosan, was significantly different from other chitosanases. To our knowledge, this is the first report to investigate the different cleavage patterns of chitosanase for COSs and cello-oligosaccharides. Key points • The molecular docking showed C-2 group of sugar units in substrate affecting the cleavage pattern. • The first chitosanase exhibited different cleavage patterns towards chito- and cello-oligosaccharides. • The groups at C-2 influence the subsite composition of the enzyme’s active cleft. Graphical abstract