title: Improved reaction pattern of an endoglycanase from Paenibacillus cookii for chitosan oligosaccharide production.
abstract: The reaction pattern of an endoglycanase from Paenibacillus cookii SS-24 (Pgl8A) was improved to facilitate chitosan oligosaccharide production. Based on the sequence alignment with chitosanase of a known structure, we performed site-directed mutagenesis of possible substrate-binding residues in Pgl8A. The mutants were expressed in Escherichia coli cells, and their cellulase and chitosanase activities were then characterised. Our results indicated that three amino acid residues (W139, W208 and E285) were important for the substrate specificity of Pgl8A. D156 and Y390 were also essential for the cellulase and chitosanase activities of Pgl8A. The products of chitosan degradation by W139A, W208A and E285Q mutants were different from those by the wild type. A chitosan pentamer accumulated following chitosan degradation by W139A, W208A and E285Q mutants. Thus, the mutants obtained in this study are potentially useful for the production of biofunctional chitosan oligosaccharides.
authors.affiliation: Department of Environmental Biology, College of Bioscience and Biotechnology, Chubu University, Kasugai, Aichi, Japan. suguru_shinoda@yahoo.co.jp
authors.forename: Suguru
authors.initials: S
authors.lastname: Shinoda
authors.forename: Shin
authors.initials: S
authors.lastname: Kanamasa
authors.forename: Motoo
authors.initials: M
authors.lastname: Arai
mesh_terms: D000595:Amino Acid Sequence; D048271:Chitosan / Q000378:metabolism*; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D006868:Hydrolysis; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009844:Oligosaccharides / Q000096:biosynthesis*; D056507:Paenibacillus / Q000201:enzymology*; D013379:Substrate Specificity
chemical_list: D009844:Oligosaccharides; D048271:Chitosan; D006026:Glycoside Hydrolases; C020668:glycanase