Production and Characterization a-Amylase Bacillus Licheniformis Recombinant In Saccharomyces Cerevisiae Using Growth Medium Containing Gliserol and Starch

• Main Author: GAYATRI (NIM 10505058), LIZA
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/12337
• Institution: Institut Teknologi Bandung

a-Amilase is a catalyst for the hydrolysis of a-D-(1,4) glycosidic bond on starch to yield oligosaccharide and dextrin. a-Amylase has many application, in a wide variety of industries such as food industry, fermentation, textile, paper, detergent, pharmaceutical, and the sugar industry. a-Amylase gene from Bacillus licheniformis (bla) had been expressed in Saccharomyces cerevisiae using growth media Yeast Extract Peptone Galactose (YPG). The bla gene expression was controlled by phosphoglycerate kinase-galactose promoter (PGK-GAL) that is induced specifically by galactose. In this study, an alternative yeast growth media, Yeast Extract Peptone Glycerol (YEPG) with the addition of starch as an inducer was used. Based on the a-amilase activities determined by Dinitrosalisicylic acid method, the specific activity obtained for a-amilase from YEPG media (2% gliserol) with the addition of starch (0.1% soluble starch) was 1.98 x 103 U / mg, while the specific activity of a-amilase recombinant in YPG medium (2% galactose) was 2.29 x 103 U / mg protein. The a-amilase from Bacillus licheniformis has a molecular mass of around 60 kDa and has a termostable characteristic with the optimum temperature of 70oC at pH 7. <br />