An Enzymatic Assay Method for Bacterial Alpha-amylase Using a Glucose Determination-kit and Maltohexaose as the Substrate

Using maltohexaose G₆ as a substrate, a convenient analytical method for amylase-catalyzed hydrolytic reaction was developed by application of an enzymatic assay-kit (glucose oxidase-peroxidase) to measurement of glucose, produced on the reaction: G₆→G₅ + G. The initial velocity was confirmed to be proportional to the initial concentration of amylase, examined, ∼ 100 μm. The new method was found in a good correlation with the Nelson and Somogyi method. Rate assay was also examined: The initial velocity is proportional to the enzyme concentration, ∼ 30 μm. Hence, this enzymatic assay method is recognized to be convenient and useful for the measurement of α-amylase.