N-Terminal Region of Chitinase I of Bacillus circulans KA-304 Contained New Chitin-Biding Domain

作者：Yano Shigekazu; Suyotha Wasana; Honda Arata; Takagi Kazuyoshi; Rattanakit Chandet Nopakarn; Wakayama Mamoru^*; Tachiki Takashi

来源：Bioscience Biotechnology and Biochemistry, 2011, 75(2): 299-304.

DOI：10.1271/bbb.100659

• family-19 chitinase
• chitin-binding domain
• protoplast

Chitinase I (CHI1) of Bacillus circulans KA-304 forms protoplasts from Schizophyllum commune mycelia when the enzyme is combined with alpha-1,3-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 (Cat CHI1), 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 CatCHI1 were lower than those of CHI1, and CatCHI1 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.

• 出版日期2011-2