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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
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author Oliveira, Hugo M.
Pinheiro, Alice Q.
Fonseca, António J.M.
Cabrita, Ana R.J.
Maia, Margarida R.G.
author_facet Oliveira, Hugo M.
Pinheiro, Alice Q.
Fonseca, António J.M.
Cabrita, Ana R.J.
Maia, Margarida R.G.
author_sort Oliveira, Hugo M.
collection PubMed
description 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.
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spelling pubmed-63705462019-02-21 Flexible and expeditious assay for quantitative monitoring of alpha-amylase and amyloglucosidase activities Oliveira, Hugo M. Pinheiro, Alice Q. Fonseca, António J.M. Cabrita, Ana R.J. Maia, Margarida R.G. MethodsX Chemistry 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. Elsevier 2019-01-24 /pmc/articles/PMC6370546/ /pubmed/30792966 http://dx.doi.org/10.1016/j.mex.2019.01.007 Text en © 2019 The Authors http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Chemistry
Oliveira, Hugo M.
Pinheiro, Alice Q.
Fonseca, António J.M.
Cabrita, Ana R.J.
Maia, Margarida R.G.
Flexible and expeditious assay for quantitative monitoring of alpha-amylase and amyloglucosidase activities
title Flexible and expeditious assay for quantitative monitoring of alpha-amylase and amyloglucosidase activities
title_full Flexible and expeditious assay for quantitative monitoring of alpha-amylase and amyloglucosidase activities
title_fullStr Flexible and expeditious assay for quantitative monitoring of alpha-amylase and amyloglucosidase activities
title_full_unstemmed Flexible and expeditious assay for quantitative monitoring of alpha-amylase and amyloglucosidase activities
title_short Flexible and expeditious assay for quantitative monitoring of alpha-amylase and amyloglucosidase activities
title_sort flexible and expeditious assay for quantitative monitoring of alpha-amylase and amyloglucosidase activities
topic Chemistry
url 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
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