title: A new method to screen polysaccharide cleavage enzymes.
abstract: The activity of polysaccharide cleavage enzymes has usually been evaluated by qualitative plate screening methods and quantitative colorimetric or chromatographic assays. The recent development of protein engineering has shown the limits of these techniques when applied to high throughput screening. Here we propose a microplate method to measure the activity of polysaccharide cleavage enzymes through small variations in viscosity. Polysaccharide solutions are co-incubated with magnetic particles in enzyme buffers. The cleavage action of polymer-degrading enzymes increases the mobility of the particles in a magnetic field, even at low levels of enzyme activities. This reproducible, sensitive technique was used to evaluate enzymatic specificity towards substrates. BioFilm indices (BFI) determined by associated software were used to follow enzyme kinetics and measure the usual variables.
authors: Clermont Université, Université Blaise Pascal, Laboratoire de Génie Chimique et Biochimique, Polytech' Clermont Ferrand, 24 avenue des Landais BP 206, 63174 Aubière cedex, France.
authors: S
authors: S
authors: Badel
authors: C
authors: C
authors: Laroche
authors: C
authors: C
authors: Gardarin
authors: E
authors: E
authors: Petit
authors: T
authors: T
authors: Bernardi
authors: P
authors: P
authors: Michaud
mesh_terms: D057075:Enzyme Assays / Q000379:methods*; D006867:Hydrolases / Q000378:metabolism*; D008190:Lyases / Q000378:metabolism*; D060526:Magnetic Fields; D010316:Particle Size; D011134:Polysaccharides / Q000378:metabolism*; D012212:Rheology; D012996:Solutions; D014783:Viscosity
chemical_list: D011134:Polysaccharides; D012996:Solutions; D006867:Hydrolases; D008190:Lyases