Production and purification of a chitinase from Ewingella americana, a recently described pathogen of the mushroom, Agaricus bisporus

• Published in: FEMS microbiology letters Vol. 157; no. 1; pp. 189 - 194
• Main Authors: Inglis, Peter W, Peberdy, John F
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 01.12.1997
• Subjects: Chitinase, EC 3.2.1.14; Ewingella americana; Hydrophobic interaction chromatography; Mushroom disease; Hydrophobic interaction chromatography; Ewingella americana; Chitinase, EC 3.2.1.14; Mushroom disease
• ISSN: 0378-1097; 1574-6968
• DOI: 10.1016/S0378-1097(97)00475-8

The chitinolytic properties of Ewingella americana, a recently described pathogen of the mushroom, Agaricus bisporus, are reported. E. americana was shown to produce chitinolytic activity in the absence of chitin and in the presence of glucose and N-acetylglucosamine, indicating constitutive synthesis by these strains. A single 33-kDa protein with chitinolytic activity was purified to homogeneity from culture filtrates, by hydrophobic interaction chromatography using a phenyl-group substituted matrix. This enzyme, by virtue of differential activity against chromogenic chitooligosaccharides and against dye-labelled soluble carboxymethylated chitin (CM-chitin-RBV), was demonstrated to be an endochitinase. Our data suggest this 33-kDa chitinase appeared to be the only chitinolytic enzyme produced by E. americana, strains of which do not grow using chitin as a carbon source. The significance of these findings in the context of mushroom disease is discussed.