title: Cloning and expression of chitinase A gene from Streptomyces cyaneus SP-27: the enzyme participates in protoplast formation of Schizophyllum commune.
abstract: Chitinase A of Streptomyces cyaneus SP-27 or chitinase I of Bacillus circulans KA-304 showed the protoplast-forming activity when combined with alpha-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 alpha-1,3-glucanase, but also to antifungal activity and powder chitin-hydrolyzing activity.
authors.affiliation: Department of Biotechnology, Faculty of Life Sciences, Ritsumeikan University, Kusatsu, Shiga, Japan.
authors.forename: Shigekazu
authors.initials: S
authors.lastname: Yano
authors.forename: Arata
authors.initials: A
authors.lastname: Honda
authors.forename: Nopakarn
authors.initials: N
authors.lastname: Rattanakit
authors.forename: Yuta
authors.initials: Y
authors.lastname: Noda
authors.forename: Mamoru
authors.initials: M
authors.lastname: Wakayama
authors.forename: Abhinya
authors.initials: A
authors.lastname: Plikomol
authors.forename: Takashi
authors.initials: T
authors.lastname: Tachiki
mesh_terms: D002688:Chitinases / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006026:Glycoside Hydrolases; D011523:Protoplasts / Q000382:microbiology*; D011994:Recombinant Proteins; D012567:Schizophyllum / Q000648:ultrastructure*; D013302:Streptomyces / Q000201:enzymology*
chemical_list: D011994:Recombinant Proteins; D006026:Glycoside Hydrolases; D002688:Chitinases