Study on Activity of ?-Amylase BaqA in Dimethyl Sulfoxide and Ionic Liquid 1-Ethyl-3-Methyl Imidazollium Bromide

• Main Author: Rheno, Timothius
• Format: Final Project
• Language: Indonesia
• Subjects: Kimia
• Online Access: https://digilib.itb.ac.id/gdl/view/38135
• Institution: Institut Teknologi Bandung

?-Amylase (E.C.3.2.1.1) hydrolyzes ?-D-(1,4)-glycosidic bond in starch resulting in linear and branch oligosaccharides. This enzyme is widely used in various industries, such as textile, detergents, sweeteners, and paper. ?-Amylase from BaqA of Bacillus aquimaris MKSC 6.2 [BaqA] has been expressed in Escherichia coliArctic express [pET30EZ-baqA]. The purpose of this research was to study the ability of ?-amylase BaqA to degrade starch in the presence of dimethyl sulfoxide DMSO and ionic liquid 1-ethyl-3-metilimidazollium bromide [EMIM]Br. SDS-PAGE analysis showed that BaqAwas expressed both as soluble and insoluble proteins. The insoluble BaqA was dissolved in urea 8M and then refolded by decreasing the urea concentration. The refolded BaqA was purified by Ni-NTA affinity column chromatography.The refolded BaqA activity has activity 5-fold of soluble BaqA activity. The activity refolded BaqA increased 200% and 133% in 1 M DMSO dan 20% (v/v) [EMIM]Br, respectively.