摘要：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 SDSPAGE;
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.
其他摘要：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.
