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Assessment of biochemical methods to detect enzymatic depolymerization of polysaccharides
Authors:C. Rigouin  C. Delbarre Ladrat  C. Sinquin  S. Colliec-Jouault  M. Dion
Affiliation:1. Laboratoire de Biotechnologie et Molécules Marines, Ifremer, rue de l’Ile d’Yeu, BP21105, F-44311 Nantes, Cedex 3, France;2. Université de Nantes, Faculté des Sciences et des Techniques, UMR-CNRS 6204, Biotechnologie, Biocatalyse, Bioregulation, 2 rue de la Houssinière, BP 92208 Nantes, F-44322 Cedex 3, France
Abstract:Biochemical methods for detection of depolymerisation were compared. Currently used methods such as reducing sugars assays, double bond monitoring or molecular weight determination were tested to follow the kinetic of depolymerization with different enzyme/polysaccharide combinations. The range of concentrations of different enzymes allowed us to identify the most sensitive and appropriate method to detect polysaccharide degradation. Reducing sugars assays are quantitative, sensitive and usable with all kind of polysaccharide but some compounds may interfere with them. When the polysaccharide is charged, agarose gel electrophoresis, although being a qualitative assay is as sensitive as high performance size exclusion chromatography analysis, easy to handle, high-throughput and this is preferred.
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