Biomolecules amylase

Amylases (EC 3.2.1.1) is an enzyme of glycoside hydrolases mainly produced in the salivary glands and pancreas, play a well-known role in hydrolyzing a-1,4- glycosidic bonds between glucose in starch (consists of two types of polysaccharide amylase and amylopectin) and maltose is release. Elevated level of Amylases in serum can be used as markers for clinical diagnosis of diseases, e.g. Pancreatitis. When the pancreas is diseased or inflamed, amylase releases into the blood.

The α -amylase activity is measured using a colorimetric method with 3,5-dinitrosalicylic acid (DNS) reagent. In this method, starch by amylase is converted into maltose. Maltose released from starch is measured by the reduction of 3,5-dinitrosalicylic acid.

Starch + H2O α-Amylase Maltose (reducing agent)

Maltose reduces the pale yellow coloured alkaline 3, 5-Dinitro salicylic acid (DNS) to the orange- red colored. The intensity of the color is proportional to the concentration of maltose present in the sample.

This intensity change in color is measured using a spectrophotometer as the absorbance at 540nm wavelength. Wave length is set to 540 nm because it is the region where orange-red color absorbs