PEMURNIAN ALPHA-AMILASE DARI CACING TANAH PERIONYX EXCAVATUS

• Main Author: Dharma, Setiawi
• Format: Final Project
• Language: Indonesia
• Subjects: Kimia
• Online Access: https://digilib.itb.ac.id/gdl/view/72239
• Institution: Institut Teknologi Bandung

This paper reports a purification of ?-amylases from earthworm Perionyx excavatus showing activity in a wide range pH between 3 and 12 and also in a wide range temperature from 30–70 oC. The ?-amylases from a gastrointestinal tract of earthworm P. excavatus were extracted using 25 mM glycine-NaOH buffer, pH 9 at 4 oC. The enzyme was further purified by the following steps: fractionation ammonium sulfate precipitation; hydrofobic interaction chromatography using phenyl sepharose column; weak ionic exchange chromatography using DEAE-sepharose column; and strong ionic exchange chromatography using Resource Q column. The ?-amylase activity was assayed by the method of Fuwa; and the protein was assayed by the method of Bradford. The result showed that the purified protein had a specific activity of 8.01 U/mg, a purification level of 176 fold, and yield 6.3%.