An improved method for detection and quantification of chitinase activities

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Abstract:

Chitinases are enzymes that serve critical roles in fungal growth and development, in resistance of plants to fungal pathogens, and in parasitism of insects by entomopathogenic fungi. The term "chitinase" is used for 3 enzymatic activities: N-acetylglucosaminidases, which sequentially release N-acetylglucosamine residues from the chitin polymer; chitobiosidases, which release disaccharides; and endochitinases, which cleave within the polymer and release oligosaccharides. We describe a technique where chitinases are separated on non-denaturing polyacrylamide gels, activities are visualized and characterized with chitinase specific substrates, and specific activities are estimated by image analysis. This technique permits a rapid determination of all of the types of chitinases present within a sample as well as their activities.Key words: chitinases, electrophoresis, non-denaturing gels.

Les chitinases sont des enzymes remplissant des rôles essentiels dans la croissance et le développement des champignons, dans la résistance des plantes aux champignons pathogènes et dans le parasitisme d'insectes par des champignons entomopathogènes. Le terme « chitinase » est utilisé pour désigner 3 activités enzymatiques: les N-acétylglucosaminidases qui libèrent séquentiellement les résidus N-acétylglucosamine du polymère de chitine; les chitobiosidases qui libèrent des disaccharides; et les endochitinases qui clivent à l'intérieur du polymère les libèrent des oligosaccharides. Nous décrivons une technique qui consiste à séparer des chitinases sur des gels de polyacrylamide non dénaturants, à visualiser les activités, à caractériser les substrats spécifiques à la chitinases et à estimer les activités spécifiques par imagerie. Cette technique permet de déterminer rapidement tous les types de chitinases présents à l'intérieur d'un échantillon de même que leurs activités.Mots clés : chitinases, électrophorèse, gels non dénaturants.[Traduit par la Rédaction]

Document Type: Research Article

Publication date: June 1, 2005

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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