Cloning and expression of chitinase A gene from Streptomyces cyaneus SP-27: The enzyme participates in protoplast formation of Schizophyllum commune

• Main Authors: Shigekazu Yano, Arata Honda, Nopakarn Rattanakit, Yuta Noda, Mamoru Wakayama, Abhinya Plikomol, 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=47949114147&origin=inward; http://cmuir.cmu.ac.th/jspui/handle/6653943832/60158
• Institution: Chiang Mai University

Chitinase A of Streptomyces cyaneus SP-27 or chitinase I of Bacillus circulans KA-304 showed the protoplast-forming activity when combined with α-1,3-glucanase of B. circulans KA-304. The gene of chitinase A was cloned. It consisted of 903 nucleotides encoding 301 amino acid residues, including a putative signal peptide (35 amino acid residues). The deduced N-terminal moiety of chitinase A showed sequence homology with the chitin-binding domain of chitinase F from Streptomyces coelicolor and chitinase 30 from Streptomyces olivaceoviridisis. The C-terminal moiety also showed high sequence similarity to the catalytic domain of several Streptomyces family 19 chitinases as well as that of chitinase I of B. circulans KA-304. Recombinant chitinase A was expressed in Escherichia coli Rosetta-gami B (DE 3). The properties of the recombinant enzyme were almost the same as those of chitinase A purified from a culture filtrate of S. cyaneus SP-27. The recombinant enzyme was superior to B. circulans KA-304 chitinase I not only in respect to protoplast forming activity in a mixture containing α-1,3-glucanase, but also to antifungal activity and powder chitin-hydrolyzing activity.