Purifikasi Parsial Enzim Karbofuran Hidrolase dari Strain Lokal Pseudomonas

Abstract

The use of pesticides has reached an alarming level in Indonesia. This study aims to purify hydrolase enzyme and to determine the highest purification value of enzyme fractionation that plays a role n the decomposition of carbofuran by the local strain of Pseudomonas. Production of extracellular and intracellular enzymes used Mineral Salt Medium (MSM) containing 100 ppm of carbofuran. The activity of the hydrolase enzyme was determined using spectrophotometry at wavelength 700 nm. Intracellular and extracellular fractionation of the carbofuran hydrolase enzyme was carried out with ammonium sulfate concentration of 30, 50 and 70% followed by dialysis 70%. The result of this study showed the level of purification obtained in crude enzyme, precipitation, 70% ammonium sulfate and 70% dialysis at 1; 0.97581; 1.24424 and 1.26764, respectively. The highest value of enzyme purification was found at a concentration of 70% ammonium sulfate with specific activity values obtained of 8.20 U/μg for extracellular enzyme and 7.75 U/μg for intracellular enzyme.