Purification of an Extracellular Amylase Produced by <i>Aspergillus niger</i> ICP2 through Submerged Fermentation

Abstract

The future applications of amylase have sparked the interest of several industries, resulting in the discovery of possible amylase-producing microbes. Aspergillus niger ICP2 was an indigenous fungus isolated from coffee pulp that shows amylolytic activity after growing on a soluble starch agar. In this study, we optimized the production and successfully purified the amylase from Aspergillus niger ICP2. Amylase optimization production was performed for a 7-day of incubation under submerged fermentation. The amylase activity was measured using the iodin method, and the total protein was quantified spectrophotometrically at 280 nm. Purifying amylase crude extract conducted such as ammonium sulfate precipitation, dialysis, and anion-exchange chromatography. Amylase was maximally precipitated at 90% saturation, followed by dialysis on a 10 kDa column, increasing specific activity up to 12.17 U/mg and purity multiples of 9.31 times. Two peaks of amylase activity were formed when the dialyzed amylase was loaded onto anion exchange chromatography with specific activities of 14.47 U/mg (fraction I) and 32.73 U/mg (fraction II). The increase in specific activity and purity fold indicated that the amylase purification process was successful.