TY - JOUR
T1 - Characterization of cellulase from E. coli BPPTCC-EGRK2
AU - Haryanto, Mas Gunawan
AU - Setyahadi, Siswa
AU - Sahlan, Muhammad
AU - Yohda, Masafumi
AU - Fukutani, Yosuke
AU - Suryono, Eko Agus
AU - Gozan, Misri
N1 - Publisher Copyright:
© 2018 Owned by the authors, published by EDP Sciences.
PY - 2018/8/27
Y1 - 2018/8/27
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85054524106&partnerID=8YFLogxK
U2 - 10.1051/e3sconf/20185200024
DO - 10.1051/e3sconf/20185200024
M3 - Conference article
AN - SCOPUS:85054524106
SN - 2555-0403
VL - 52
JO - E3S Web of Conferences
JF - E3S Web of Conferences
M1 - 00024
T2 - 2018 Consortium Studies of Smallholder Palm Oil International Conference, CSSPO 2018
Y2 - 9 July 2018 through 11 July 2018
ER -