Characterization of cellulase from E. coli BPPTCC-EGRK2

Abstract

Cellulase enzymes are widely used in various industries such as detergent industry, bioethanol, animal feed, textile and paper. This research focused on characterization of cellulase produced from Eschericia coli BPPT-CC EgRK2, which is a recombinant that can produce protein enzymes endo- β-1,4-glucanase. Eschericia coli BPPT-CC EgRK2 was cultured in 1 litre liquid medium Luria Bertani. Because the bacteria is intracellular, sonication is needed for cell disruption to get the cellulase enzyme. The enzyme activity was then analyzed by CMC substrate at different concentration. The protein content analysis was carried out using Bradford method; the molecular weight analysis was done using SDS-PAGE; while the enzyme kinetics was plotted using Michaelis-Menten model. Our results showed the highest enzyme activity was 2.403 U/ml and the protein concentration was 5.352 mg/ml. The Michaelis-Menten constant (Km) and maximum velocity (Vmax) for CMC substrate hydrolysis were 0.07 μmol/ml and 2.49 μmol/ml/sec, respectively. The cellulase molecular weight was 58 kDa using SDS-PAGE with 7.5% stacking gel. The results indicated that Eschericia coli BPPT-CC EgRK2 is a promising renewable source for cellulase production for industrial application.

Progression of Recombinant Cellulolytic and Ethanologenic Escherichia coli for Production of Bioethanol: A Review

“…Subsequently, further manipulation of substrate concentrations and the enzyme kinetics of the reactions can be studied. Enzyme activity measures the functionality of the cellulases whereas constants in enzyme kinetics reveal the relationship between substrate and cellulase affinity for each other, maximum velocity of cellulase reaction rate at substrate saturation phase and the turnover number of the enzymes [4]; [15]; [16]; [17]; [18]; [19].…”

Optimasi Selulase pada Enzyme Assisted Extraction Lemak dari Caulerpa Lentillifera Menggunakan Response Surface Methodology

“…Konsentrasi substrat (g/L) kemudian diplotkan dengan kecepatan reaksi (μmol/menit) untuk membentuk kurva. Perhitungan kecepatan reaksi menjadi bentuk glukosa dengan melibatkan massa molekul relatif (Mr) glukosa yaitu sebesar 180 (Haryanto et al, 2018). Kecepatan reaksi yang dihasilkan yaitu 0 µmol/menit -14,1 µmol/menit (Gambar 1).…”