Characterization of cellulase from E. coli BPPTCC-EGRK2

Mas Gunawan Haryanto, Siswa Setyahadi, Muhamad Sahlan, Masafumi Yohda, Yosuke Fukutani, Eko Agus Suryono, Misri

Research output: Contribution to journalConference articlepeer-review

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.

Original languageEnglish
Article number00024
JournalE3S Web of Conferences
Volume52
DOIs
Publication statusPublished - 27 Aug 2018
Event2018 Consortium Studies of Smallholder Palm Oil International Conference, CSSPO 2018 - Sarawak, Malaysia
Duration: 9 Jul 201811 Jul 2018

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