α-amylase production from Endomyces fibuliger – an indigenous yeast isolate of Western Himalayas

Amylase production, its purification, characterization and optimization have been studied in Endomyces fibuliger. The enzyme was purified to about 13-fold by using gel permeation chromatography (Sephadex G-25, Sephadex G-100). The purity of enzyme was checked by the use of Native-PAGE and SDS-PAGE. The molecular weight of enzyme was 55KD with the presence of two polypeptide units. Enzyme was most active at pH 7.0, temperature 30°C with optimum incubation time of 30 minutes. The maximum activity of the enzyme was at 2% (w/v) soluble starch concentration. The enzyme was para-constitutive in nature. The parameters for amylase production were optimized by Response Surface Methodology (RSM). The best carbon source was soluble starch, the optimum pH values were 5.0 and 8.0, and the optimum temperatures were 30°C and 37°C. Ca+² and Mg+² had enhancing effect on its production.