Critical review on conventional spectroscopic α‐amylase activity detection methods: merits, demerits, and future prospects

Abstract

α‐Amylase is an endoenzyme that catalyses the hydrolysis of internal α‐l,4 glycosidic linkages in polysaccharides to produce maltose, maltotriose, and α‐limit dextrins. It is widely used in the laboratorial and industrial workflow for several applications. There are several methods utilizing different techniques and substrates to assess α‐amylase activity, among which the spectroscopic methods have found widespread applicability due to their ease of use and cost‐effectiveness. Depending upon the reaction principle, these assays are classified into four groups: reducing sugar, enzymatic, chromogenic, and amyloclastic methods. Despite the presence of numerous methods, there is no general reliable method to assess α‐amylase activity. Each method is shown to have its own merits and demerits. Many improvements have been made to make the available methods more accurate, reliable, and easy. This communication briefly discusses the basic reaction mechanisms and critically reviews the advantages and shortcomings associated with each method. Further recommendations are made for future development. © 2020 Society of Chemical Industry

High‐performance thin‐layer chromatography coupled attenuated total reflectance‐Fourier‐transform infrared and NMR spectroscopy‐based identification of α‐amylase inhibitor from the aerial part of Asparagus racemosus Willd

“…Through the breakdown of internal glycosidic linkages, α-amylase catalyses the hydrolysis of starch and similar polysaccharides into glucose as well as oligosaccharides. 32 Iodine reacts with amylose in starch to generate a dark blue or purple complex, although it does not react with glucose. 33 In the presence of iodine, the starch that was not hydrolysed by α-amylase due to enzyme inhib- showed lowest antioxidant activity (4.29 μg GAE/μL; Table 3).…”

Production of extracellular α-amylase by single-stage steady-state continuous cultures of Candida wangnamkhiaoensis in an airlift bioreactor

“…Amylases (α, β, and γ) are among the most versatile and important families of enzymes that have numerous biotechnological applications [ 1 , 2 ]. Among them, α-amylases (α-1,4-glucan-4-glucanohydrolase; E.C.…”

“…For instance, food, brewery, syrups, textile, detergent, pulp, paper, pharmaceutical, environmental, biofuel, healthcare, clinical, and analytical chemistry industries, among others [ 1 , 3 , 4 ]. Nowadays, α-amylases hold the major world market share of enzymes, accounting for approximately 25–30% of the world´s enzyme production [ 2 , 3 ].…”

Inhibitory properties of polyphenols in Malus “Winter Red” crabapple fruit on α‐glucosidase and α‐amylase using improved methods

“…The characteristic absorption at 540 nm could be used to indicate enzyme activity (Visvanathan et al, 2020). However, most dietary polyphenols have reducing potential and could react with DNS, leading to an overestimation of reducing sugar content, and thus the activity of α-amylase (Shahidi & Ambigaipalan, 2015).…”