Purification and Characterization a-Amylase Bacillus licheniformis (BLA) SITH in Pichia pastoris

• Main Author: ALDILA (NIM 10504052), SEPTIN
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/11394
• Institution: Institut Teknologi Bandung

a-Amylase is an enzyme which hydrolyses a-1,4 glycosidic linkages of the polysaccharides into soluble maltooligosaccharide, maltodextrin, and small quantity of glucose. a-Amilase has been used in various industries, such as starch processing, textile, and bakery. Thermostable a-amylase needed for liquifation process in starch processing industry. a-Amylase Bacillus licheniformis (BLA) is a thermostable a-amylase. A commercial production of a-amylase in industry requires an efficient cell factory capable of producing a-amylase at very high level. Pichia pastoris is methylotropic yeast which has been used for commercial production of many enzymes. Aims of the research were to purify and characterize of recombinant B. licheniformis a-amylase (BLA.SITH1) produced in P. pastoris. Purification of recombinant BLA.SITH1 by affinity chromatography Ni-NTA (His-tagged Purification). The recombinant BLA.SITH1 has an optimum temperature of 65 oC and it has T50 of 80 minutes at pH 6 and temperature 65oC. Thin Layer Chromatography (TLC) analysis showed that the major product hydrolysis starch are G2 (maltose), G3(maltotriose) and G5 (maltopentose) which indicates that recombinant BLA.SITH1 is an endo-acting enzyme. <br />