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Flexible and expeditious assay for quantitative monitoring of alpha-amylase and amyloglucosidase activities

The monitoring of the activity of alpha-amylase and amyloglucosidase is an important tool for studying their role in the hydrolysis of starch. Here we introduced an improved method capable to measure the activity of alpha-amylase and amyloglucosidase from different sources based on a quantitative st...

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Detalles Bibliográficos
Autores principales: Oliveira, Hugo M., Pinheiro, Alice Q., Fonseca, António J.M., Cabrita, Ana R.J., Maia, Margarida R.G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6370546/
https://www.ncbi.nlm.nih.gov/pubmed/30792966
http://dx.doi.org/10.1016/j.mex.2019.01.007
Descripción
Sumario:The monitoring of the activity of alpha-amylase and amyloglucosidase is an important tool for studying their role in the hydrolysis of starch. Here we introduced an improved method capable to measure the activity of alpha-amylase and amyloglucosidase from different sources based on a quantitative starch-iodine assay. The developments of the assay sought the consistent preparation of the reagents, the rescale of the assay and the adjustment of the sensitivity. This was complemented by a glucose yield assay for amyloglucosidase that allowed a secondary source of information when insoluble starches were studied. The proposed method showed high precision in long-term use (RSD < 6.3%). Furthermore, all experimental conditions can be adapted according to the equipment available at each laboratory, transforming this method in a broadband analytical tool for screening alpha-amylase and amyloglucosidase activities. • Tailorable assay based on the starch-iodine staining for the determination of alpha-amylase and amyloglucosidase activities. • Enhanced consistence of reagent preparation. • High intra-day and inter-day reproducibility.