Molecular characterization and antifungal activity of a family 46 chitosanase fromAmycolatopsissp. CsO-2
| Autor: | Yoshiho Nagata, Akikazu Ando, Daisuke Miyatsuchi, Takeshi Watanabe, Akihiro Saito, Takaaki Ooya, Kanako Terakado, Shin-ya Nakayama, Hiroko Fuchigami |
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| Rok vydání: | 2009 |
| Předmět: |
Antifungal Agents
Glycoside Hydrolases Rhizopus oryzae Amycolatopsis Microbiology Streptomyces Fungal Proteins Mutant protein Actinomycetales Escherichia coli Genetics Chitosanase Cloning Molecular Molecular Biology Peptide sequence Chitosan Chitosanase activity biology Chitinases Sequence Analysis DNA biology.organism_classification Recombinant Proteins Biochemistry Mutation Mutagenesis Site-Directed Streptomyces griseus Rhizopus |
| Zdroj: | FEMS Microbiology Letters. 293:79-84 |
| ISSN: | 1574-6968 0378-1097 |
| DOI: | 10.1111/j.1574-6968.2009.01507.x |
| Popis: | An actinomycete strain, Amycolatopsis sp. CsO-2, produces a 27-kDa chitosanase. To reveal the molecular characteristics of the enzyme, its corresponding gene ctoA was cloned by a reverse genetic technique, based on the N-terminal amino acid sequence of the protein. The encoded CtoA protein was deduced to be composed of 286 amino acids, including a putative signal peptide (1-48), and exhibited 83% identity in the amino acid sequence with the family 46 chitosanases from Streptomyces sp. N174 or Nocardioides sp. N106. The active recombinant CtoA protein was successfully overproduced in Escherichia coli. The mutant protein E22Q, in which the glutamic acid residue 22 was replaced with glutamine, abolished the chitosanase activity, showing that the Glu22 residue is required for the enzymatic activity. CtoA exhibited antifungal activity against Rhizopus oryzae, which is known to produce chitosan probably as a cell wall component. In contrast, E22Q did not inhibit the growth of the fungus, suggesting that chitosan-hydrolyzing activity is essential for the antifungal activity. It is noteworthy that the antifungal effect of CtoA against R. oryzae was drastically enhanced by the simultaneous addition of the family 19 chitinase ChiC from Streptomyces griseus. |
| Databáze: | OpenAIRE |
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