E3S Web Conf.
Volume 52, 2018CSSPO International Conference 2018: Towards Inclusive & Sustainable Agriculture – Harmonizing Environmental, Social and Economic Dimensions: Is it Possible?
|Number of page(s)||7|
|Published online||27 August 2018|
Characterization of cellulase from E. coli BPPTCC-EGRK2
Chemical Engineering Department, Faculty of Engineering, Universitas Indonesia,
2 Center for Bioindustrial Technology, Agency for Assessment and Application of Technology, 15314 Serpong, Indonesia
3 Department of Biotechnology and Life Science Tokyo University of Agriculture and Technology 2-24-16 Naka-cho, Koganei, Tokyo Japan 184-8588
4 Faculty of Biology, Gadjah Mada University, Indonesia
* Corresponding author: firstname.lastname@example.org
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.
© Owned by the authors, published by EDP Sciences, 2018
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