CRYSTALLIZATION OF β-AMILASE ENZYME FROM CILEMBU SWEET POTATOES (Ipomoea batatas cv. Cilembu)

• Main Author: AMELIA HANDAYANI (NIM : 10508059); Pembimbing I : Dr. Fida Madayanti W Pembimbing II : Dr. Ih, CITRA
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/15237
• Institution: Institut Teknologi Bandung

&#946;-amylase is an exoamylase enzyme which hydrolyzes &#945;-1,4-glycosidic bonds at non-reducing end of polysaccharide, producing maltose units. Maltose syrup contains high concentration of maltose and becomes more popular nowadays as a diabetic diet. Protein structural study through purification and crystallization of &#946;-amylase plays an important role in developing &#946;-amylase applications, such as making maltose syrup from starch. One of the &#946;-amylase sources is Cilembu sweet potatoes. Objective of this research was purification and crystallization of &#946;-amylase from Cilembu sweet potatoes. In order to obtain the research objective, purification was carried out with several steps, isolation of crude enzyme extract, addition of excess substrate (1500 mg/mL), fractionation by ammonium sulphate, and ion exchange chromatography DEAE-Sephacel. The purity of &#946;-amylase increased 28 and 82 times for F0&#8722;60% and F60&#8722;80%, respectively after the ion exchange chromatography DEAE-Sephacel. Hydrolysis products of enzyme identified by Thin Layer Cromatography (TLC) showed that the main product was maltose. Based on polyacrylamide gel electrophoresis analysis, it obtained a single &#946;-amylase band with size of 256 kDa (native-page) and 64 kDa (SDS-page). This results implied that the &#946;-amylase was an homotetramer enzyme. Pure amylase was then crystallized by commercial kit crystallization and showed some tiny crystals. <br /> <br />