Karakter Enzim Amilase dari Bacillus licheniformis Teramobil dengan Variasi Matriks Kalsium Alginat

Abstract

Enzim amilase Bacillus licheniformis berperan dalam pemecahan pati dalam berbagai aplikasi industri, namun stabilitas dan efisiensinya belum optimal. Upaya untuk meningkatkan performa enzim dilakukan melalui amobilisasi pada matriks kalsium alginat (Ca-alg). Penelitian ini mengkaji karakter enzim amilase dari B. licheniformis teramobil dengan variasi matriks kalsium alginat (2-4%). Ekstrak kasar enzim dimurnikan dengan fraksinasi amonium sulfat 80% dan dialisis, kemudian diamobilisasi dengan Ca-alg. Bobot molekul, aktivitas, dan kadar protein enzim amilase amobil diukur dengan metode SDS-PAGE, reduksi DNS (3,5-Dinitrosalicylic acid), dan Lowry. Aktivitas enzim dipengaruhi oleh berbagai faktor, antara lain konsentrasi substrat, suhu, pH, dan waktu inkubasi. Aktivitas optimal tercapai pada enzim amobil 2% pada konsentrasi pati 2%, suhu 60 °C, pH 6, dan waktu inkubasi 40 menit. Kadar protein menurun setelah proses pemurnian dan amobilisasi. Data SDS-PAGE menunjukkan bahwa bobot molekul amilase berkisar antara 55-70 kDa. Efisiensi amobilisasi tertinggi (75,28%) dicapai pada enzim amobil 2% dan menurun dengan peningkatan konsentrasi Ca-alg.

Amylase enzyme from Bacillus licheniformis plays a role in starch degradation in various industrial applications, however stability and efficiency remain suboptimal. Efforts to enhance enzyme's performance were carried out through immobilization on a calcium alginate (Ca-alg) matrix. This study examines characteristics of immobilized amylase enzyme from B. licheniformis with varying calcium alginate matrix (2-4%). Crude enzyme extract was purified using 80% ammonium sulfate fractionation and dialysis, followed by immobilization with Ca-alg. Molecular weight, activity, and protein content of immobilized enzyme were measured using SDS-PAGE, DNS (3,5-Dinitrosalicylic acid) reduction, and Lowry. Enzyme activity was influenced by various factors, including substrate concentration, temperature, pH, and incubation time. Optimal activity was achieved with 2% immobilized enzyme at 2% starch concentration, 60 °C temperature, pH 6, and 40 minutes of incubation time. Protein content decreased after purification and immobilization. SDS-PAGE data showed that molecular weight of amylase ranged from 55-70 kDa. Highest immobilization efficiency (75.28%) was achieved with 2% immobilized enzyme and decreased with higher Ca-alg concentrations.