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

灵活、快速地定量监测 α-淀粉酶和淀粉葡萄糖苷酶活性

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作者:Hugo M Oliveira, Alice Q Pinheiro, António J M Fonseca, Ana R J Cabrita, Margarida R G Maia

Abstract

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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