N-terminal region of chitinase I of bacillus circulans KA-304 contained new chitin-biding domain

• Main Authors: Shigekazu Yano, Wasana Suyotha, Arata Honda, Kazuyoshi Takagi, Nopakarn Rattanakit-Chandet, Mamoru Wakayama, Takashi Tachiki
• Format: Journal
• Published: 2018
• Subjects: Biochemistry, Genetics and Molecular Biology; Chemistry; Immunology and Microbiology
• Online Access: https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=79952164185&origin=inward; http://cmuir.cmu.ac.th/jspui/handle/6653943832/49733
• Institution: Chiang Mai University

Chitinase I (CHI1) of Bacillus circulans KA-304 forms protoplasts from Schizophyllum commune mycelia when the enzyme is combined with a-l,3-glucanase of B. circulans KA-304. CHI1 consists of an N-terminal unknown region and a C-terminal catalytic region classified into the glycoside hydrolase family-19 type. An N-terminal region-truncated mutant of CHI 1 (CatCHIl), which was expressed in Escherichia coli Rosetta-gami B (DE3), lost colloidal chitin- and powder chitin-binding activities. The colloidal chitin- and the powder chitin-hydrolyzing activities of CatCHIl were lower than those of CHI1, and CatCHIl was not effective in forming the protoplast. A fusion protein of the N-terminal region of CHI1 and green fluorescent protein (Nterm-GFP) was expressed in E. coli, and the fusion protein was adsorbed to colloidal chitin, powder chitin, and chitosan. Fluorescence microscopy analysis showed that Nterm-GFP bound to the S. commune cell-wall.