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Abstrak :
Mengetahui potensi selulase dalam mendegradasi dinding kista Acanthamoeba sp.Sampel Acanthamoeba sp.didapatkan dari isolat koleksi Departemen Parasitologi FKUI yang mana 2 sampel berasal dari pasien dan 1 sampel dari lingkungan. Ketiga sampel dikultur dengan menggunakan media non-nutrien agar NNA dan diidentifikasi dengan PCR dan sekuensing. Kemudian dilakukan optimasi konsentrasi dan waktu inkubasi selulase dengan jumlah kista yang digunakan 5 x 103. Kandidat konsentrasi selulase yang digunakan adalah 50 U, 100 U, 150 U, 200 U, 250 U, dan 300 U dengan waktu inkubasi yang digunakan adalah 2 jam, 4 jam, 6 jam, 8 jam, dan 24 jam. Selanjutnya hasil perlakuan dengan konsentrasi dan waktu inkubasi yang paling optimal diamati dengan menggunakan SEM untuk melihat perubahan permukaan dinding kista. Kemudian dilakukan uji kistasidal untuk mengetahui efektifitas kistasidal larutan desinfektan, selulase dan campuran larutan desinfektan dan selulase dalam membunuh kista Acanthamoeba sp.dinilai berdasarkan nilai viabilitasnya.Konsentrasi selulase yang paling optimal dalam membunuh kista Acanthamoeba sp.adalah 300 U dengan waktu inkubasi 24 jam. Persentase viabilitas Acanthamoeba sp.yang diberi paparan larutan desinfektan saja selama 24 jam adalah 95 , selulase saja selama 24 jam 75 , dan campuran selulase dan larutan desinfektan selama 24 jam adalah 25 . Selulase mampu mendegradasi dinding kista Acanthamoeba sp.Konsentrasi selulase yang optimal dalam mendegradasi dinding kista Acanthamoeba sp.adalah 300 U dengan waktu inkubasi yang optimal 24 jam. Penambahan selulase ke larutan desinfektan berpotensi untuk meningkatkan efektivitas larutan desinfektan karena selulase mampu mendegradasi dinding kista sehingga memungkinkan larutan desinfektan untuk masuk dan membunuh kista Acanthamoeba sp.

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The goal of this study is to know the potential of cellulase in degradation of cyst wall Acanthamoeba sp.Sample of Acanthamoeba sp. obtained from isolate collection of Department of Parasitology FKUI which 2 samples come from patient and 1 sample from environment. All three samples were cultured using non nutrient agar NNA media and identified by PCR and sequencing. The concentration of cellulase concentration used was 50 U, 100 U, 150 U, 200 U, 250 U, and 300 U with the incubation time used was 2 hours , 4 hours, 6 hours, 8 hours, and 24 hours. Furthermore, treatment results with the most optimum concentration and incubation time were observed by using SEM to see changes in the surface of the walls of the cyst. Then performed cysticidal test to determine the effectiveness cysticidal of disinfectant solution, cellulase, and combination of disinfectant solution and cellulase in killing Acanthamoeba sp. cyst assessed by their viability value. The most optimal cellulase concentration in killing Acanthamoeba sp.cysts. is 300 U with incubation time of 24 hours. Percentage of viability of Acanthamoeba sp.which was exposed to a disinfectant solution for 24 hours was 95 , cellulase alone for 24 hours 75 , and the combination of cellulase and disinfectant solution for 24 hours was 25 . Cellulase are capable of degrading Acanthamoeba sp.cyst wall. Optimal cellulase concentration in degrading Acanthamoeba sp. cyst wall is 300 U with an optimal incubation time is 24 hours. The addition of cellulase to the disinfectant solution has the potential to increase the effectiveness of the disinfectant solution because cellulase are capable of degrading the cyst wall allowing the disinfectant solution to enter and kill Acanthamoeba sp.cysts.

Abstrak :
ABSTRAK
Enzim selulase digunakan secara luas dalam berbagai industri, namun hampir 99% kebutuhan enzim domestik dipenuhi oleh impor. Salah satu biomassa lignoselulostik yang memiliki potensi tinggi produksi selulase adalah Tandan Kosong Sawit (TKS) karena kandungan selulosanya mencapai 41,3 – 46,5% (w/w). Metode konvensional untuk produksi enzim selulase adalah menggunakan jamur, namun diketahui bahwa bakteri juga dapat memproduksi enzim selulase dengan laju yang lebih cepat. Dalam penelitian ini dikaji bagaimana produksi enzim selulase oleh isolat BPPTCC-RK2 dan rekombinan EgRK2 dengan melakukan penentuan waktu inkubasi, suhu operasi dan pH optimum. Didapat bahwa waktu untuk produksi terbesar selulase untuk kedua jenis isolat adalah 24 jam, dengan pH dan suhu optimum untuk rekombinan EgRK2 adalah 7 dan 40 C. Setelah ekstraksi enzim, diperoleh nilai pH dan suhu optimum untuk selulase dari BPPTCCRK2 sebesar 6,5 dan 50 C, sementara selulase dari EgRK2 adalah sebesar 6,5 dan 60 C. Nilai Km dan Vmax selulase dari BPPTCC-RK2 untuk degradasi CMC adalah sebesar 0,021% dan 1,631 mol.ml-1min 1 dan dari rekombinan EgRK2 adalah sebesar 0,097% dan 2,739 mol.ml-1min-1. Sementara, nilai Km dan Vmax selulase pada degradasi TKKS adalah sebesar 0,704% dan 0,943 mol.ml-1min-1 untuk BPPTCC-RK2 dan 0,26% dan 1,934 mol.ml-1min-1 untuk rekombinan EgRK2.

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ABSTRACT
Cellulase enzyme is widely used in industries, however almost 99% of Indonesia industrial enzyme demands is fulfilled by imports. One of the lignocellulosic biomass which has pretty high cellulose content is Oil Palm Empty Fruit Bunch (OPEFB), which reaches up to 41,3 – 46,5% (w/w). The conventional method for cellulase production is by utilizing fungi, but it is known that cellulase also can be produced by bacteria with higher production This research examined how was the production of cellulase enzyme by Bacillus amyloliquefaciens BPPTCCRK2 and EgRK2 recombinant by varying incubation time, operating temperature and medium pH. It was known that the optimum time for cellulase production by both isolates was 24 hours, with optimum pH and temperature of 7 and 40 C respectively. After enzyme extraction, the optimum pH and temperature of cellulase were obtained, with the value of 6,5 and 50 C for BPPTCC-RK2 and 6,5 and 60 C for EgRK2 recombinant. Km and Vmax value for CMC degradation were 0,021% and 1,631 mol.ml-1min 1 for BPPTCC-RK2 and 0,097% and 2,739 mol.ml-1min-1 for EgRK2 recombinant. Meanwhile, Km and Vmax value for OPEFB degradation were 0,704% and 0,943 mol.ml-1min-1 for BPPTCC-RK2 0,26% and 1,934 mol.ml-1min-1 for EgRK2 recombinant.

Abstrak :
Jerami padi merupakan salah satu limbah lignoselulosa pertanian yang jumlahnya cukup melimpah dan mengandung komponen lignin, selulosa, dan hemiselulosa yang dapat dimanfaatkan sebagai bahan baku/substrat yang digunakan untuk pembuatan selulase, sehingga memiliki nilai ekonomi dan ramah lingkungan. Sebelum lignoselulosa digunakan sebagai substrat perlu dilakukan minimalisasi kadar ligninnya dengan menggunakan pretreatment kimia basa dengan menggunakan NaOH 4%. Kapang yang digunakan adalah Trichoderma viride strain T051, jamur ini merupakan penghasil enzim selulase yang berfungsi menghidrolisis selulosa menjadi glukosa. Karakteristik enzim selulase berdasarkan mekanisme hidolisis ada tiga jenis, yaitu endoglukanase, exoglukanase dan glukosidase. Aktivitas enzim dengan menggunakan substrat jerami yang didelignifikasi basa lebih tinggi dibandingkan dengan jerami tanpa delignifikasi. Variasi nutrisi pada medium produksi yang memberikan unit aktivitas optimum adalah dengan penambahan medium basal pada substrat uji aktivitas CMC 1% sebesar 59,97 mU/mL. Definisi satu unit aktivitas adalah 1 μmol glukosa yang dihasilkan permenit pada suhu 450C. Enzim dengan aktivitas tertinggi selanjutnya difraksionasi menggunakan amonium sulfat dengan kenaikan tingkat kejenuhan dan didialisis. Hasil penelitian menunjukkan fraksi amonium sulfat dengan kejenuhan 50-70% memiliki aktivitas tertinggi sebesar 62,55 mU/mL dengan aktivitas spesifik sebesar 16,96 mU/mL. Hasil dialisis memiliki aktivitas spesifik sebesar 24,94 mU/mg. pH optimum aktivitas enzim selulase adalah 5. Logam Cu2+ dapat menginhibisi aktivitas enzim selulase, sementara Zn2+ dan Mg2+ memberi dampak peningkatan aktivitas enzim. ......Rice straw is one of lignocellulosic agricultural waste which is quite abundant and contain components of lignin, cellulose, and hemicellulose which can be used as raw materials / substrates used to manufacture cellulase, so it has economic value and environmental friendliness. Before the lignocellulose is used as the substrate is necessary to minimize the levels of lignin using alkaline chemical pretreatment using NaOH 4%. Fungus that used were Trichoderma viride strain T051, this fungus is a producer of cellulase enzymes that function hydrolyze cellulose into glucose. Characteristics of cellulase enzymes by mechanisms hidolisis there are three types, namely endoglukanase, exoglukanase and glucosidase. Enzyme activity by using straw substrate base delignification higher than the straw without delignification. Variation of nutrients in the medium production unit that provides an optimum activity is the addition of basal medium on the substrate 1% CMC activity assay of 59.97 mU/mL. Definition of one unit of activity is 1 mol of glucose produced per minute at 450C. With the next highest enzyme activity fractionated using ammonium sulfate with increasing levels of saturation and dialyzed. The results showed fractions with ammonium sulfate saturation of 50-70% has the highest activity of 62.55 mU/mL with a specific activity of 16.96 mU/mL. The results of dialysis had a specific activity of 24.94 mU/mg. The optimum pH of the enzyme activity of cellulase is 5. Metals Cu2+ can inhibit cellulase enzyme activity, whereas Zn2+ and Mg2+ gives the impact of increased enzyme activity.

Abstrak :
ABSTRAK
Penelitian ini bertujuan untuk mendapatkan enzim selulase dari kapang terpilih untuk pembuatan selulosa mikrokristal dari kulit buah kapuk. Alfa selulosa didapatkan melalui biodelignifikasi dan enzim selulase murni diperoleh dari galur kapang terpilih. Selulosa mikrokristal didapatkan melalui hidrolisis enzimatis dengan enzim selulase yang telah dimurnikan, lalu diidentifikasi dengan analisa kualitatif Fourier transformed infrared spectroscopy (FTIR), dan differential scanning calorimetry (DSC), diikuti oleh karakterisasi selulosa mikrokristal seperti x-ray diffraction (XRD), analisis ukuran dan distribusi partikel, dan pengukuran scanning electron microscope-energy dispersive x-ray (SEM-EDX). Hasil penelitian menunjukkan bahwa biodelignifikasi terbaik dilakukan pada suhu 40˚C menghasilkan14,88% α-selulosa. Penicillium sp. sebagai kapang terpilih memiliki aktivitas selulase tertinggi dengan indeks selulolitik 4,83 dan aktivitas selulase sebesar 0,04299 U/mL. Fraksi pertama digunakan untuk hidrolisis memiliki aktivitas tertinggi yaitu 649,68 mU/mL. Hasil identifikasi FTIR menunjukkan kemiripan diagram dengan Avicel PH 101 dengan titik lebur 244,580˚C. Karakterisasi XRD menunjukkan kristalinitas pada 2 puncak 2Ɵ (deg) nilai 22,58 dengan intensitas 634 dan nilai 21,85 dengan intensitas 51. Susut pengeringan 3,74%, derajat keasaman pH 7,0, ukuran partikel antara 13,06 hingga 196,79μm, kerapatan serbuk ruah 0,111 g/cm3, serbuk mampat 0,235 g/cm3, laju alir cukup baik, SEM-EDX menunjukkan bentuk morfologi selulosa mikrokristal kulit buah kapuk berbentuk memanjang. Selulosa mikrokristal kulit buah kapuk telah menunjukkan karakteristik yang berbeda dengan referensi dan dapat dikembangkan lebih lanjut.

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ABSTRACT
This study aims to obtain cellulase enzymes from selected molds for microcrystalline cellulose preparation from α-cellulose of kapok pericarpium. Alpha cellulose was obtained by biodelignification and the purified cellulase was obtained from selected mold. The Microcrystalline cellulose that obtained from enzymatic hydrolysis then identified by qualitative analysis, Fourier transformed infrared spectroscopy (FTIR), and differential scanning calorimetry (DSC), followed by characterization of microcrystalline cellulose includes X-Ray Diffraction (XRD), Particle Size and Distribution Analysis (PSA), and Scanning Electron Microscope-Energy Dispersive X-ray (SEM-EDX). Biodelignification carried out at a temperature of 40C produced 14.88% α-cellulose, Penicillium sp. as the selected mold had the highest cellulase activity with a cellulolytic index of 4.83 and cellulase activity of 0.04299 U/mL. The first fraction used for hydrolysis had the highest activity of 649.68 mU/mL. FTIR identification showed a similarity with Avicel PH 101 with a melting point of 244.580C. XRD characterization was showed the crystallinity at 2 peaks 2Ɵ (deg) 22.58 with intensity 634 and 21.85 with intensity 51. Loss on drying was 3.74%, pH was 7.0, particle size ranged from 13.06 to 196.79 um, bulk density and tapped density were 0.111 g/cm3 and 0.235 g/cm3, the flow rate character is quite good, and SEM-EDX was showed that the morphological shape of the microcrystalline cellulose of the kapok pericarpium is elongated. Microcrystalline cellulose has shown a difference in characteristic and can be furthered.

Abstrak :
Penggunaan bahan bakar fosil oleh manusia menimbulkan ancaman serius, yaitu jaminan ketersediaan bahan bakar fosil untuk beberapa dekade mendatang dan polusi akibat emisi pembakaran bahan bakar fosil ke lingkungan. Kesadaran terhadap ancaman tersebut telah mengintensifkan berbagai riset yang bertujuan menghasilkan sumber-sumber energi alternatif yang berkelanjutan dan lebih ramah lingkungan. Salah satu energi alternatif yang relatif murah ditinjau aspek produksinya dan relatif ramah lingkungan adalah pengembangan bioetanol dari limbah kertas yang banyak mengandung lignoselulosa. Penelitian pembuatan etanol dari kertas intinya adalah dengan proses Sakarafikasi dan Fermentasi Serentak (SSF). Enzim selulase dan yeast Saccharomyces cerevisiae digunakan untuk hidrolisis dan fermentasi dalam proses SSF tersebut. PH yang digunakan adalah pH 5 karena pada penelitian konversi etanol sebelumnya pH 5 adalah pH optimum. Proses SSF dilakukan dengan waktu inkubasi selama 6, 12, 24, 36, 48, 72, 96 jam. Aktifitas yang digunakan adalah 0,2; 0,3; dan 0,5gr. Sebelum dilakukan proses hidrolisis dan fermentasi perlu adanya proses Pada penelitian ini jenis limbah kertas yang diuji adalah hanya limbah kertas HVS bertinta, HVS kosong dan kertas koran. Penelitian konversi limbah kertas menjadi etanol dengan dengan menggunakan enzim selulase yang akan dilakukan diharapkan mampu membantu riset-riset selanjutnya dan dikembangkan ke arah komersial untuk mendukung konservasi energi dan penggunaan energi alternatif bioetanol sebagai pensubstitusi minyak bumi yang ketersediaannya mulai terbatas, serta diharapkan limbah-limbah khususnya limbah kertas yang menjadi permasalahan bagi beberapa Negara dapat tertangani dengan baik. Hasil penelitian menunjukkan bahwa etanol tidak dapat dihasilkan tanpa enzim selulase. Pada kertas HVS kosong kandungan selulosanya adalah sekitar 60,5 %, pada HVS bertinta kandungan selulosanya adalah sekitar 58,3 %, dan pada kertas koran kandungan selulosanya sekitar 49,1%. Pada kertas HVS bertinta, HVS kosong dan kertas koran diperoleh konsentrasi etanol tertinggi berturut-turut 1238,9 ppm, 669 ppm, 1428 ppm.

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The use of fossil fuel by humans threatens serious problems for the future such as the availability of fossil fuel for further decades and pollution to the environment by emissions from the burning of fossil fuel. Awareness of these problems has increased the intensity of research to produce an alternative energy resource that is both sustainable and environmentally friendly. One example of a sustainable alternative energy resource that is relatively cheap and environmentally friendly is the development of bio-ethanol from waste paper that contains large amounts of lignocelluloses. The point of this research deals with the production of ethanol from paper using the simultaneous process of saccharification and fermentation (SSF.) The Cellulose enzyme and Saccharomyces cerevisiae were used to hydrolyse and ferment during the SSF process. The pH level used was pH 5 because previous research on ethanol conversion had shown that pH 5 is the optimum level. The SSF process was done with an incubation period of 6, 12, 24, 36, 48, 72, and 96 hours. The activity used was 0.2, 0.3, and 0.5gr. Before the hydrolyse and fermenting processes are done we need another process (''') For this research the type of waste paper tested was HVS paper with ink, blank HVS paper and newspaper. Research about converting waste paper to ethanol using the cellulose enzyme will hopefully be used to help future research and commercial development to support energy conservation and the use of the alternative bio-ethanol as a substitute for a limited supply of oil. Also we hope that garbage specifically waste paper which has become a problem for so many countries can be handled in a positive way. The results of this research show that blank HVS paper's cellulose content is around 60.5%, HVS paper with ink has a cellulose content of 58.3% and with newspapers the content is around 49.1%. In regards to blank HVS paper, HVS paper with ink, and newspaper, the highest ethanol concentration in succession is 1238.9ppm, 669 ppm, and 1428 ppm.

Abstrak :
The bioetanol development from biomass bases of lignocellulose like bagasse is one of alternative energy which has potential to be applied in Indonesia. Beside of raw material source that is so many in our country, the process is also environmentally friendly. Conversion of bagasse becomes etanol using Simultaneous Sacharification and Fermentation (SSH technology by cellulose and cellobiase enzyme had been done on this research. Sacharification process or hydrolysis process, cellulose enzyme will break cellulose polymer becomes glucose whereas cellobiose enzyme will break cellobiose becomes glucose. Then, glucose through fermentation is changed to etanol by using yeast Saccharomyces cerevisiae. The variations include pH of system that is pH 4' ; 4,5 and 5, HCI addition low concentrated HCI at pH 5 with variation of concentration that is 0,5 % and I %, also variation of sample at pH 5 where bagasse without pretreatment is compared with bagasse which had been done pretreatment by using fungi Lentinus edodes for 4 weeks. The result shows that the use of cellulose and cellobiase enzyme with system optimum condition pH 5 produce etanol concentration is higher than using only cellulose enzyme at the same pH condition. For substrate concentration 50 g/L, on the use of cellulose and cellobiase, the highest etanol concentration which is produced bagasse without pretreatment is 5,62 g/L or li,24 % from bagasse. On HCI addition, the highest etanol concentration is produced by concentration HCI i % with amount 6,52 g/L or 13,04 % from bagasse. With bagasse L. edodes and P. ostreatus 6 weelts, the highest etanol concentration that is 6 86 g/L and 6,50 g/L or 13, 72% and l2,99% from bagasse. It also shows that HCl addition low concentrated and pretreatment by white rot fungi L. edodes and P. ostreatus can increase the etanol quantity that is produced from bagasse conversion.

Abstrak :
ABSTRAK
Enzim selulase banyak digunakan dalam berbagai industri seperti industri deterjen, bioethanol, pakan ternak, tekstil dan kertas. Akan tetapi saat ini kebutuhan enzim selulase paling banyak didapat dari impor. Salah satu bakteri penghasil enzim selulase adalah Eschericia coli BPPT-CC EgRK2. Bakteri hasil rekombinasi yang dapat memproduksi enzim protein endo- 𝜷-1,4-glukanase, diteliti di dalam kultur batch untuk ditentukan parameter-parameter kinetikanya seperti konstanta Michaelis-Menten (Km) dan kecepatan maksimum (Vmax). Eschericia coli BPPT-CC EgRK2 dikultur di dalam media cair Luria Bertani. Selanjutnya dilakukan purifikasi dan karakterisasi dari enzim selulase. Purifikasi menggunakan metode kromatografi penukar ion dan filtrasi gel setelah itu dianalisis aktifitas enzim dengan substrat carboxymethyl cellulose (CMC), kadar protein menggunakan metode Bradford, berat molekul menggunakan sodium deodecyl sulfate (SDS-PAGE) dan kinetika enzim menggunakan plot Michaelis-Menten. Hasilnya menunjukkan aktivitas enzim tertinggi adalah 3.114 U/ml dan konsentrasi selulase 0.723 mg/ml. Konstanta Michaelis-Menten (Km) dan kecepatan maksimum (Vmax) untuk hidrolisis substrat CMC adalah 0.314 μmol/ml and 3.511 μmol/ml/sec. Hasil dari analisis berat molekul selulase menggunakan metode SDS-PAGE adalah 58 kDa pada 7.5% stacking gel. Hasil dari penelitian ini membuktikan bahwa Eschericia coli BPPT-CC EgRK2 menjadi sebuah sumber terbarukan dari enzim selulase untuk aplikasi pada skala industrial

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ABSTRACT
Cellulase enzymes are widely used in various industries such as detergent industry, bioethanol, animal feed, textile and paper. This research is focused on characterization cellulase enzyme from bacteria. One of the bacteria producing cellulase enzyme is Eschericia coli BPPT-CC EgRK2. Recombinant bacteria that can produce protein enzymes endo- β-1,4-glucanase. Eschericia coli BPPT-CC EgRK2 is cultured in 1 litre liquid medium Luria Bertani. Because the bacteria is intracellular, need sonication to break the cell to get the cellulase enzyme. Then purification with ion exchange chromatography and gel filtration to purified the enzyme. After that analyzed the enzyme activity with carboxymethyl cellulose (CMC) substrate at different concentration, protein content analysis using Bradford method, molecular weight analysis using sodium deodecyl sulfate (SDS-PAGE) and enzyme kinetics using Michaelis-Menten plot. This results showed the highest enzyme activity is 3.11 U/ml at 2% CMC and the cellulase concentration is 0.723 mg/ml. The Michaelis-Menten constant (Km) and maximum velocity (Vmax) for CMC substrate hydrolysis is 0.314 μmol/ml and 3.511 μmol/ml/sec. The results of cellulae enzyme molecular weight is 58 kDa using SDS-PAGE with 7.5% stacking gel. The result of this research have known that Eschericia coli BPPT-CC EgRK2 become a promising renewable source for cellulase enzyme for industrial application.

Abstrak :
Microbial protease and cellulase are in high demand by different industries due to their minimal cost and availability. This study was aimed to maximize the production of protease and cellulase using two bacteria, Corynebacterium alkanolyticum ATH3 and Bacillus licheniformis CBH7, isolated from fish gut. This study demonstrated the effect of different culture parameters in protease and cellulase production using two different bacterial strains. Results of this study clearly indicated the importance of different parameters such as moisture content, pH, incubation temperature, incubation period, inoculum size, carbon sources and nitrogen sources in enzyme production. The most critical parameters affecting the enzymes production were pH, temperature, carbon and nitrogen sources. Further investigations are required to enhance the enzymes production using genetic engineering.

Abstrak :
Potensi keanekaragaman Indonesia memberikan peluang untuk mendapatkan mikroorganisme penghasil endo-β-1,4-glukanase yang mampu menghidrolisis selulosa. Bacillus amyloliquefaciens BPPTCC-RK2 telah berhasil diisolasi dari rayap. Gen endo-β-1,4-glukanase dikloning dari DNA genom B.amyloliquefaciens BPPTCC-RK2 menggunakan metode rekombinatorial dan diekspresikan secara fungsional di dalam E.coli. Didapatkan ORF sepanjang 1500 nukleotida yang menyandikan 499 asam amino dengan berat molekul 55 kDa. Gen dikloning kedalam pDEST14 dan dioverekspresi pada E.coli BL21-Star. Aktivitas tertinggi sebesar 26,05 U/mg protein setelah diinduksi dengan 1mM IPTG selama 24 jam. Enzim optimum pada pH 6,0 dan suhu 65℃ dan memiliki waktu paruh 90 menit pada suhu 60℃. Pada konsentrasi etanol 100 g/L, masih memberikan aktivitas hingga 78% setelah 24 jam. ...... Indonesia has potential biodiversity that provides opportunities to obtain endo-β-1,4-glucanase producing microorganism that could hydrolize cellulose. Bacillus amyloliquefaciens BPPTCC-RK2 have been isolated from termites. Endo-β-1,4-glucanase gene have been cloned from genomic DNA B.amyloliquefaciens BPPTCC-RK2 using recombinatorial method and functionally expressed in E.coli. A full length gene of endo-β-1,4-glucanase consisting 1500 nucleotides that encoded for a protein 499 amino acids with predicted molecular weight 55 kDa. Highest enzyme activity (26,05 U/mg) achieved after 24 hour induction with 1mM IPTG. The enzyme optimum at pH 6,0 and temperature 65℃ and 90 minutes half-life at 60℃. This enzyme give 78% residual activity after 24 hour incubation in 100 g/L ethanol.