The Synergy of Recombinant Xylanolytic Enzyme on Xylan Hydrolysis

Microbial xylanases or xylanolytic enzyme have received considerable attention over the last years owing to a multitude of  possible applications. These  enzymes  have  potential  in  the  biodegradation of lignocellulosic biomass  to  fuels, chemicals, fruit juice, animal feed and in improving rumen digestion. More recently, the use of xylanases as bleaching agent  in  the  pulp  and  paper  industry  has  been  suggested  to  replace  of some  of  the  chemicals  presently  used  for  this purpose. Such applications could  have an important positive impact on the environment. The purpose of  this research was  determining  the  synergy  of  3 recombinant  xylanolytic  enzymes  (β-xylosidase,  exo-xylanase  and α-L-arabinofuranosidase)  from  recombinant  Eschericia  coli  BL21 (DE-star)  in  xylan  hydrolysis  by  analysis  the  reduction sugar product. Purified of recombinant xylanolytic enzyme β-xylosidase  (Xyl), exo-xylanase  (Exo-Xyl) and α-L-arabinofuranosidase  (Abfa)  with  Ni-NTA resin.  Seven  samples  of  enzyme  (each  and  enzyme  mixture)  used to hydrolyze  xylan    substrate  (oat-spelt  xylan).  Analysis  of  hydrolysis product  was  done  by  HPLC.  The  xylanolytic activities of this enzyme before and after purification were 0,91 and 9,94 U/mL (Exo-Xyl); 1,65 and 14,2 U/mL (Xyl); 0,65  and  5,6  U/mL  (Abfa).  The  xylosidase  activity  were  2,37  and  14,3  U/mL  (Xyl);  1,49  and  10,5  U/mL  (Exo-Xyl); 2,54  and  18,6  U/mL  (Abfa).  The  highest  hydrolysis  product  of  xylan  (xylose)  shown  in  enzyme  mixture  of    exo-xylanase and β-xylosidase was 1,084 mg/mL. &nbsp

THE SYNERGY OF RECOMBINANT XYLANOLYTIC ENZYME ON XYLAN HYDROLYSIS

Microbial xylanases or xylanolytic enzyme have received considerable attention over the last years owing to a multitude of  possible applications. These  enzymes  have  potential  in  the  biodegradation of lignocellulosic biomass  to  fuels, chemicals, fruit juice, animal feed and in improving rumen digestion. More recently, the use of xylanases as bleaching agent  in  the  pulp  and  paper  industry  has  been  suggested  to  replace  of some  of  the  chemicals  presently  used  for  this purpose. Such applications could  have an important positive impact on the environment. The purpose of  this research was  determining  the  synergy  of  3 recombinant  xylanolytic  enzymes  (β-xylosidase,  exo-xylanase  and α-L-arabinofuranosidase)  from  recombinant  Eschericia  coli  BL21 (DE-star)  in  xylan  hydrolysis  by  analysis  the  reduction sugar product. Purified of recombinant xylanolytic enzyme β-xylosidase  (Xyl), exo-xylanase  (Exo-Xyl) and α-L-arabinofuranosidase  (Abfa)  with  Ni-NTA resin.  Seven  samples  of  enzyme  (each  and  enzyme  mixture)  used to hydrolyze  xylan    substrate  (oat-spelt  xylan).  Analysis  of  hydrolysis product  was  done  by  HPLC.  The  xylanolytic activities of this enzyme before and after purification were 0,91 and 9,94 U/mL (Exo-Xyl); 1,65 and 14,2 U/mL (Xyl); 0,65  and  5,6  U/mL  (Abfa).  The  xylosidase  activity  were  2,37  and  14,3  U/mL  (Xyl);  1,49  and  10,5  U/mL  (Exo-Xyl); 2,54  and  18,6  U/mL  (Abfa).  The  highest  hydrolysis  product  of  xylan  (xylose)  shown  in  enzyme  mixture  of    exo-xylanase and β-xylosidase was 1,084 mg/mL.  Keywords: α-L-arabinofuranosidase, β-xylosidase, exo-xylanase, xylan, xylos

Potential of Phytase Enzymes as Biocatalysts for Improved Nutritional Value of Rice Bran for Broiler Feed

ABSTRACT Phytase is one of the enzymes belonging to the group phosphatase capable of hydrolyzing phytic compounds in the form of myo-inositol (1.2.3.4.5.6) hexsa into myo-inositol phosphatase and organic phosphate . In the digestive tract nonruminant livestock (poultry) there is no phytaseenzymes , it causes the content of the rice bran phytate compounds are difficult to digest because of the strong chelating properties , so it wasted phytate with feces. Restrictions on the use of rice bran in the diet because the fiber content and high phyticacid . One alternative to reduce the phytate content of the feed is to use phytase enzyme produced from Actinobacillus sp and Bacillus pumilus) is expected to hydrolyze phytic acid ( myo-inositol 1.2.3.4.5.6-hexakisphosphate) for rice bran orthophosphate in organic produce and a series of lower phosphoric into myo -inositol -free , so all minerals such as P, Cawhich is an important mineral to be released and used for the growth of broiler chickens. The results obtained adding the enzyme phytase can improve the nutritional quality of rice bran content is increasing crude protein, crude fiber and increasing decline in the availability of minerals calcium and phosphorus

Karakterisasi enzim fitase asal bakteri rumen (Actinobacillus sp dan Bacillus pumilus ) dan analisis SEM terhadap perubahan struktur permukaan dedak padi untuk ransum ayam broiler

Fitase merupakan salah satu enzim yang tergolong dalam kelompok phosphatase yang mampu menghidrolisis senyawa fitat berupa myo-inositol (1,2,3,4,5,6) hexsa phosphatase menjadi myo-inositol dan phosphat organik. Salah satu altematif untuk menurunkan kandungan fitat dalam pakan adalah dengan menggunakan bakteri penghasil enzim fitase. Tim Peneliti telah berhasil memperoleh bakteri asal rumen ruminansia (Actinobacillus sp dan Bacillus pumilus) yang diharapkan mampu menghasilkan enzim fitase sebagai Feed Supplement untuk menghasilkan bahan pakan teroak berkualitas ditinjau dari ketersediaan protein dan mineml P, Mg, Mn, Fe, Zn, Ca yang tinggi sehingga selain dapat memenuhi kebutuhan nutrisi bagi ayam broiler juga ramah lingkungan. Tujuan peneilitian ini adalah untuk mendapatkan karakterisasi enzim fitase produksi bakteri Actinobacillus sp dan Bacillus pumilus (kurva pertumbuhan bakteri, optimasi pH dan suhu, stabilitas pH dan suhu). Metode Penelitian Tabap I (Tahun I) adalah pengamatan terhadap kurva pertumbuhan bakteri Actinobacillus sp dan Bacillus pumilus, uji aktivitas enzim, karakterisasi enzirn fitase pada berbagai suhu maupun pH serta stabilitas enzim. Hasil optimasi fitase dari bakteri Actinobacillus sp diperoleh pada suhu 45°C dengan aktivitas 0,1374 UfmL, optimasi pH 4 dengan aktivitas 0,1374 UfmL. Stabilitas suhu enzirn fitase dari bakteri Actinobacillus sp cukup baik dimana enzim ini stabil pada suhu 45°C selama 2-4 jam (0,0428-0,0657 UfmL). Stabilitas pH enzim fitase dati bakteri Actinobacillus sp cukup baik dimana enzim ini stabil pada pH 4 (0,0353 UfmL). Hasil penelitian optimasi fitase dari bakteri Bacillus pumilus diperoleh pada suhu 50°C dengan aktivitas 0,0780 UfmL, optimasi pH 6 dengan aktivitas 0,0577 U/mL. Stabilitas suhu enzirn fitase dari bakteri Bacillus pumilus cukup baik dimana enzim ini stabil pada suhu 50°C selama 2-10 jam (0,0592-0,0593 UfmL). Stabilitas pH enzim fitase dari bakteri Bacillus pumilus cukup baik dimana enzim ini stabil pada pH 4-6 (0,0216-0,0351 U/mL). Disimpulkan enzim fitase bakteri Actinobacillus sp memptmlai prosentase pH dan suhu optimum masing-masing 4 dan 45° C, stabil pada suhu 45 C selama 6 jam dengan rentang pH 4 6. Enzim fitase bakteri Bacillus pumilus mempunyai prosentase pH dan suhu optimum masing-masing 6 dan 50° C, stabil pada suhu 50° C selama 10 jam dengan rentang pH 4 6

Lignocellulosic Enzymes Characterization and Scanning Electron Microscope Analysis on Rice Bran Surface Structure Changes

Rice bran was one of livestock food ingredients wit h high crude fibers contents, including cellulose and hemicelluloses. Lignocellulosic enzymes such as xylanase and cellulase functioned in degrading cellulose and hemicelluloses. The objectives of this study were to qualitatively screen Actinobacillus sp. (G6) and Bacillus pumilus (G7) which produced xylanase and cellulase, determining molecular weight of xylanase and cellulase enz ymes using SDS-PAGE and Zymogram, and analyzing structure changes on rice bran surface after Scanning Electron Microscope treatments using xylanase and cellulase enzymes. The findings of this study indicated xylanolitic indexes for G6 and G7 were 2 and 1.7 respectively while cel lulosic index for G6 and G7 were 1.6 and 2. Enzymesproduced by Actinobacillus sp. (G6) had molecular weight 25- 35 kDa and above 100 kDa while enzymes produced by Bacillus pumilus (G7) had molecular weight 25 kDa up to above 100 kDa. The result of rice bran SEM analysis indicated profile change of the samples before and after enzymes treatmen

Hidrolisis beberapa jenis xilan dengan enzim xilanolitik termofilik rekombinan

The aims of this research were to know the ability of recombinant xylanolytic enzyme from recombinant E. coli DH5a (pTP510) to hydrolyze several commercial xylan and analysis the reduction sugar product. Recombinant xylanolytic enzyme (exo-xylanase, b-xylosidase and a-L-arabinofuranosidase) could hydrolyzed several commercial xylan (oat-spelt xylan, birchwood, wheat, rye, and arabinan) with xylanolytic activities are: oat-spelt xylan (1.73 U/mL), birchwood (0.92 U/mL), wheat (6.52 U/mL), rye (4.94 U/mL), and arabinan (3.40 U/mL). Xylanolytic enzyme assay use specific substrate p-nitrophenyl-b-D-xylopyranoside (pNP-X) shown xylosidase activity 15.869 U/mL. Hydrolysis product was analyzed by HPLC. The results showed that xylose, arabinose, and xylo-oligosaccharide were produced from birchwood, wheat, rye, and arabinan hydrolysis, although xylose and arabinose were produced from hydrolysis of oat-spelt xylan

Characterization of Fungal Laccase Isolated from oil palm empty fruit bunches (OPEFB) and Its Degradation from The Agriculture Waste

In recent years, many studies have been developed to overcome the abundance of lignocellulosic waste. Laccase is a metalloenzyme that can degrade lignin. This enzyme has useful properties that make it applicable for the agricultural sector in Indonesia. The present study is designed to isolate and determine the activity of laccase found in the fungi of oil palm empty fruit bunch (OPEFB), and the fungal laccase was precipitated and partially purified by using ammonium sulfate. The optimum temperature of fungal laccase was 40 °C for fungus lacc-B and 50 °C for fungus lacc-A, C, and D. The optimum pH was obtained 5.0 for fungus lacc-A and D, pH 6.0 for fungus lacc-C and pH 7.0 for fungus lacc-B. Furthermore, the activity of crude extract were 1.4 U.mL−1, 1.5 U.mL−1, 1.3 U.mL−1, and 1.3 U.mL−1 for fungus lacc-A, B, C, and D. Purification levels of fungus lacc-A, B, C, and D increased 1.7, 1.6, 1.9, and 2.5 times, respectively after ammonium sulfate precipitation and dialysis. Fungus lacc-D has the highest level of laccase activity than other fungi. Physical analysis of the agricultural waste after fungus lacc-D addition indicated that the surface profile is damaged, hollow, and broken. The delignification from corn cob and rice straw was observed with scanning electron microscope (SEM) which showed the presence of morphological changes in the lignocellulose waste sample