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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...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2019
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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. |
format | Online Article Text |
id | pubmed-6370546 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
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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