Purification and Characterization of Extracellular, Polyextremophilic α-amylase Obtained from Halophilic Engyodontium album

Abstract

Background: a-Amylases (EC 3.2.1.1) are covering approximately 25% of total enzyme market and are frequently used in food, pharmaceutical and detergent industries. Objectives: The first ever detailed characterization of amylase from any halophilic Engyodontium album is presented. Materials and Methods: An extracellular α-amylase was studied from halophilic E. album TISTR 3645. The enzyme was extracted and purified by column chromatography. SDS-PAGE was performed to find the molecular weight of the enzyme. The effects of pH, temperature and salinity on the isolated enzyme were determined. The effects of various additives on enzyme were studied. Results: The molecular weight of the amylase was 50 kDa. The enzyme specific activity was 132.17 U.mg-1 with Vmax and Km values of 15.36 U.mg-1 and 6.28 mg.ml-1, respectively. The optimum enzyme activities were found at pH 9.0, 60ºC and 30% (w/v) NaCl. BaCl2, CaCl2, HgCl2 and MgCl2 improved amylase activity. b-mercaptoethanol, EDTA, FeCl2 and ZnCl2 decreased enzyme activity. Conclusions: Polyextremophilic characteristics of a-amylase from halophilic E. album TISTR 3645 were revealed during the characterization studies, demonstrating promising features, making it a useful candidate for various industries.