Pemurnian dan Karakterisasi Enzim Glukosa Oksidase dari Isolat Lokal Aspergillus niger (IPBCC.08.610)

Abstract

Production of various enzymes using microorganisms is one way of developing microorganism biodiversity in Indonesia. For this study, glucose oxidase (GOD) which is widely available for use and purchase in the commercial market was used. This research intended to determine the optimum incubation time for the production of GOD from Aspergillus niger local isolate (IPBCC.08.610) and determine the specific activity of the crude extract and purified enzymes fraction. In addition, it sought to determine the optimum temperature and pH level in GOD reaction and establish the values of Km and vmax. Optimum incubation time for GOD production of A. niger (IPBCC.08.610) was 48 hours with initial velocity of the reaction and the specific activity at 0.80/minute and 3.21 U/mg. Ammonium sulfate with 80% saturation was optimum to precipitate the GOD protein with specific activity of the fraction at 22.18 U/mg and purity levels at 6.92 times. Specific activity of the fraction of dialysis was 21.25 U/mg and the level of purity was 6.63 times. Km and vmax values were 46 mM and 11 U/mg with optimal temperature and pH level for GOD reaction between 37 - 45°C and pH 5.5.