Detection of alginate lyase by activity staining after sodium dodecil sulfate-polyacrylamide gel electrophoresis and subsequent renaturation.
A slab gel electrophoretic method for the study of bacterial alginate lyase has been developed. By incorporating alginate in acrylamide gels, the method is based on renaturation of the enzyme after polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate, subsequent staining with cetylpyridinium chloride, and quantification of the spot by densitometric scanning. The molecular mass of alginate lyase can be determined from its position in the gel.
Peciña
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Instituto de Bioquimica Vegetal y Fotosíntesis, Universidad de Sevilla-Consejo Superior de Investigaciones Científicas, Spain.
Paneque
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D001395:Azotobacter / Q000201:enzymology; D001426:Bacterial Proteins / Q000032:analysis* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D007700:Kinetics; D011133:Polysaccharide-Lyases / Q000032:analysis* / Q000378:metabolism; D011489:Protein Denaturation; D012680:Sensitivity and Specificity; D012967:Sodium Dodecyl Sulfate; D013194:Staining and Labeling / Q000379:methods
D001426:Bacterial Proteins; D012967:Sodium Dodecyl Sulfate; D011133:Polysaccharide-Lyases; C016158:poly(beta-D-mannuronate) lyase