Production and characterization of cellulase from E. coli EgRK2 recombinant based oil palm empty fruit bunch

Amraini, Said Zul; Ariyani, Lina Putri; Hermansyah, Heri; Setyahadi, Siswa; Rahman, Siti Fauziyah; Park, Don-Hee; Gozan, Misri

doi:10.1007/s12257-017-0034-2

Cited by 25 publications

(17 citation statements)

References 25 publications

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“…Luria Bertani (LB) solution was used as a standard medium for enzyme production with composition per litre are 10 g peptone, 5 g yeast extract and 5 g NaCl. This solution was added with 1% cellulose [11,17]. The medium was then dissolved by water and sterilized in an autoclave at 121°C and 1.2 atm for 15 minutes.…”

Section: Enzyme Productionmentioning

confidence: 99%

Optimization of cellulase production by bacillus sp. BPPT CC RK2 with pH and temperature variation using response surface methodology

et al. 2018

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Indonesia has abundant ethanol biomass feedstocks. However the second-generation ethanol production process is still hampered by the unavailability of cellulase enzyme in the process of decomposition of lignocellulose into saccharides that can be processed into ethanol through fermentation. Cellulase is known as exozyme produced by Bacillus sp. in submerged fermentation. In this study, cellulase production by Bacillus sp. CC BPPT RK2 on natural and abundant agricultural waste substrates (rice bran and coconut water) was evaluated by investigating the optimum conditions for cellulase production in a 50 ml laboratory scale. Preliminary test using Luria Bentani (LB) medium with additional CMC (1%) were done to select optimum range of pH and Temperature. The preliminary tests results were then followed by optimization of pH and temperature, which were carried out using response surface methodology (RSM). RSM optimization model showed optimum values 6.23 for pH and 40.04 °C, with 14 terms (each with 1 degree of freedom), 4 linear effects, 6 interaction effects and 4 quadratic effects. These optimization by RSM results were slightly different compared to preliminary test, showing the effect of interactions between parameters. The characteristics of interaction among variables tested against the cellulase activity are reported in this study including: positive effects on cellulase activity of the resulting responses; negative interactions affecting the response of cellulase activity; synergistic interaction; and antagonistic interactions between each other.

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Section: Enzyme Productionmentioning

confidence: 99%

Optimization of cellulase production by bacillus sp. BPPT CC RK2 with pH and temperature variation using response surface methodology

et al. 2018

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“…Furfuryl alcohol is one of the most widely used furfural derivatives. Furfural production generally commences with the initial hydrolysis of the hemicellulose fraction of lignocellulose into pentose, which is then dehydrated into furfural [17]. In this process, in the batch mode, a sulfuric acid catalyst reacts with the biomass at temperatures of 170-185°C to obtain a furfural yield of approximately 40-50%.…”

Section: Introductionmentioning

confidence: 99%

“…Many parameters, such as temperature, acid concentration, and biomass characteristics, influence the rate of LA and furfural reactions. Previous studies have investigated the kinetic reactions of LA and furfural production in various types of biomass using sulfuric acid catalysts [16][17][18][19][20][21][22][23][24][25][26]. Chin et al [20] performed a kinetic study of POEFB acid hydrolysis, which produced xylose decomposition products and LA.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of Separate and Simultaneous Kinetic Parameters for Levulinic Acid and Furfural Production from Pretreated Palm Oil Empty Fruit Bunches

Gozan

Panjaitan

Tristantini

et al. 2018

International Journal of Chemical Engineering

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Palm oil empty fruit bunches (POEFBs) can be converted into levulinic acid (LA) and furfural, which are among the top building-block chemicals. The purpose of this study was to investigate separate and simultaneous kinetic model parameters for LA and furfural production from POEFBs, which were pretreated by soaking in aqueous ammonia (SAA). The highest LA yield, which was obtained at a reaction temperature of 170°C after 90 min in an acidic solution with a concentration of 1 M, was 52.1 mol%. The highest furfural yield was 27.94 mol%, which was obtained at a reaction temperature of 170°C after 20 min in an acidic solution with a concentration of 0.5 M. SAA pretreatment affected activation energy in glucose degradation reactions and favoured direct conversion of hemicellulose to furfural. The activation energy of LA production (EakHMF) increases with higher acid catalyst concentration, and the activation energy of furfural production (EakXYN) decreases with higher acid concentration. These trends in the activation energy occurred in both separate and simultaneous kinetic models. Simultaneous kinetic model is better to calculate kinetic parameters of LA and furfural production than separate kinetic models because the simultaneous kinetic model had a lower sum of square error (SSE) when estimating kinetic parameters.

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“…The gene that expressed endo-β-1.4-glucanase (gene attB-EgRK2) in Bacillus sp. RK2 was isolated and amplified by the Polymerase Chain Reaction (PCR) method.In this study we used GatewayR (Invitrogen) method and two primers such as egrk2F: 5'-AAA AAG CAG GCT CGA TGA AAC GGT CAA TYT Y -3'; egrk2R: 5'-AGA AAG CTG GGT ACT AAT TKG KTT CTG WTC CC -3' and attB1 adapter: 5'-G GGG ACA AGT TTG TAC AAA AAA GCA GGC T -3'; attB2 adapter: 5'-GGG GAC CAC TTT GTA AA GAA AGC TGG GT -3' [11].…”

Section: Sample Collectionmentioning

confidence: 99%

“…This study examined an E. coli EgRK2 recombinant strain. The recombinant strain was used in this experiment because the result of the previous research showed the wild type has low activity , the recombinant strain grows faster on inexpensive media, ease of isolation and purification of proteins excreted in comparison with the wild type [11].This study examined the effects the enzyme's hydrolysis performance on Carboxymethyl Cellulose (CMC) as the substrate. The aim of the present research is to describe the biosynthesis, isolation and characterization of a cellulase from E. coli EgRK2 isolate.…”

Section: Introductionmentioning

confidence: 99%

Characterization of cellulase from E. coli BPPTCC-EGRK2

et al. 2018

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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.

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Production and characterization of cellulase from E. coli EgRK2 recombinant based oil palm empty fruit bunch

Cited by 25 publications

References 25 publications

Optimization of cellulase production by bacillus sp. BPPT CC RK2 with pH and temperature variation using response surface methodology

Optimization of cellulase production by bacillus sp. BPPT CC RK2 with pH and temperature variation using response surface methodology

Evaluation of Separate and Simultaneous Kinetic Parameters for Levulinic Acid and Furfural Production from Pretreated Palm Oil Empty Fruit Bunches

Characterization of cellulase from E. coli BPPTCC-EGRK2

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