PJB-2024-346
INDUSTRIALLY APPLICABLE α-AMYLASE PRODUCTION, PURIFICATION AND CHARACTERIZATION THROUGH INDEGINOUS BACILLUS SUBTILIS
Muhammad Imran
Abstract
The α-amylase is an enzyme that catalyzes the hydrolysis of starch and glycogen. It is produced from plants, animals and microbes but microbial α-amylase has highest commercial and industrial value. Bacillus subtilis was identified on the basis of 16S rRNA. Different carbon and nitrogen sources were supplemented to the LB medium. The supplemented LB medium was sterilized and used for the growth of Bacillus subtilis. High yield of α-amylase was produced at 3% molasses and 3% peptone. Alpha Amylase was precipitated at 60% ammonium sulphate concentration then partially purified α-amylase was further purified through column chromatography. The molecular weight of α-amylase was determined by SDS-PAGE and size found to be ~40kDa. The purified α-amylase utilized for characterization studies. The optimum temperature and pH for α-amylase activity was found to be 55°C and 9 respectively. Enzyme kinetic studies were performed to check Km and Vmax. Values. Impact of different metal ions were observed on α-amylase activity and maximum alpha amylase activity was recorded in the presence of Fe2+. The role of α-amylase was analyzed in detergent industry with different company detergents. In textile industry, enzyme activity was estimated by checking release of reducing sugars. The enzyme capability with detergents and release of reducing sugars in de-sizing step in textile industry makes alpha amylase suitable for local industries in Pakistan.
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