Produksi, Karakterisasi, Dan Pemurnian Enzim Glukosa Oksidase Dari Isolat Aspergillus Niger IPB CC 08 610

Abstract

Glucose oxidase is an enzyme widely applied in the food, clinical, and biosensor industries. This study was conducted to produce and partially purify glucose oxidase from Aspergillus niger IPB CC 08 610 isolate and to determine its enzymatic activity and prediction Km using the ABTS-HRP assay. Enzyme extraction was performed through mechanical cell disruption employing quartz sand. Partial purification was carried out by ammonium sulfate precipitation at 80% saturation. The Km parameters of the enzyme were determined based on Michaelis–Menten analyses. The crude extract fraction yielded 38 mL of glucose oxidase with a total activity of 62.10 U, total protein of 8.61 mg, specific activity of 7.21 U/mg, a yield of 100%, and a purification fold of 1. Precipitation with 80% ammonium sulfate produced 1.2 mL of enzyme solution with a total activity of 14.09 U, total protein of 1.44 mg, specific activity of 9.75 U/mg, a yield of 22.68%, and a purification fold of 1.35. The Km value of the crude extract fraction was estimated to be 5 mM

Glukosa oksidase merupakan enzim yang banyak dimanfaatkan dalam bidang pangan, klinis, dan biosensor. Penelitian ini bertujuan menghasilkan dan memurnikan secara parsial enzim glukosa osksidase dari isolat A. niger IPB CC 08 610, serta mengukur aktivitas dan prediksi nilai Km enzim menggunakan metode ABTS-HRP. Isolasi dilakukan dengan metode penghancuran sel secara mekanik menggunakan pasir kuarsa. Pemurnian parsial dilakukan dengan metode pengendapan amonium sulfat 80%. Parameter Km ditentukan melalui analisis kurva Michaelis-Menten. Hasil penelitian diperoleh enzim glukosa oksidase pada fraksi ekstrak kasar sebanyak 38 mL dengan aktivitas total sebesar 62,10 U, protein total sebanyak 8,61 mg, aktivitas spesifik sebesar 7,21 U/mg, yield 100%, dan faktor kemurnian 1. Fraksi hasil pengendapan amonium sulfat 80% menghasilkan larutan sebanyak 1,2 mL dengan aktivitas total sebesar 14,09 U, total protein sebesar 1,44 mg, aktivitas spesifik sebesar 9,75 U/mg, yield 22,68%, dan faktor kemurnian 1,35. Nilai Km pada fraksi ekstrak kasar diprediksi sebesar 5 mM