PERBAIKAN FREENESS DAN MUTU KERTAS BEKAS MENGGUNAKAN CELLULOSE BINDING DOMAIN DARI ENDOGLUKANASE Egl-II

The objective of this research is improving the freeness of waste paper stock and paper quality by using the Cellulose Binding Domain (CBD) of endoglucanase Egl-II. CBD has been separated from endoglucanase Egl-II by proteolysis method. CBD has a molecular weight of approximately 21 kD. The produced CBD contained 0.04 mg / mL protein and did not show the total enzyme activity. Waste paper disintegrated using Niagara beater with no load at the consistency of 1.5%. CBD was applied to the refined waste paper fibers with a freeness of 200 mL CSF (Canadian Standard Freeness). The dosages of CBD used for waste paper treatment were 0.2 and 0.3 mg CBD/g of oven-dried pulp. The result shows that this treatment increases the freeness of fibers by 140 mL CSF (70%). CBD also increases the amount of removed water from the fibers from 290 mL to 390 mL and 370 mL, respectively, using the dynamic drainage jar (DDJ) measurement. The cellobiose assay of the waste paper filtrate treated with CBD shows no sugar dissolution, which indicates no cellulose degradation. The tear index of paper produced by treatment with CBD shows insignificant change. The Concora Medium Test (CMT) of paper produced by treatment with CBD has higher tensile index, burst index, and ring crush.Keywords: cellulose-binding domain, endoglucanase Egl-II, freeness improvement, waste paperABSTRAKTujuan penelitian ini adalah untuk memperbaiki freeness dari stok kertas bekas dan mutu lembaran kertas yang dihasilkan dengan menggunakan Cellulose Binding Domain (CBD) dari endoglukanase Egl-II. CBD yang digunakan merupakan hasil pemisahan dari endoglukanase Egl-II dengan metode proteolisis. CBD ini memiliki berat molekul sekitar 21 kD. CBD yang dihasilkan mengandung kadar protein sebesar 0,04 mg/mL dan tidak terdeteksi adanya aktivitas total enzim. Kertas bekas diuraikan dengan menggunakan Niagara beater tanpa beban pada konsistensi 1,5%. CBD diaplikasikan pada serat kertas bekas yang telah digiling dan memiliki freeness 200 mL CSF (Canadian Standard Freeness). Dosis CBD yang digunakan untuk perlakuan terhadap serat kertas bekas adalah 0,2 dan 0,3 mg CBD/g pulp kering-oven. Hasil penelitian menunjukkan perlakuan dengan CBD meningkatkan freeness bubur serat kertas bekas sebesar 140 mL CSF (70%). CBD juga meningkatkan volume air yang dihilangkan dari serat kertas bekas dari 290 mL menjadi 390 mL dan 370 mL menggunakan pengukuran dynamic drainage jar (DDJ). %FPR meningkat dari 98,80% menjadi 99,77%. Pengujian selobiosa terlarut pada filtrat serat kertas bekas yang telah mengalami perlakuan dengan CBD memperlihatkan tidak ada gula terlarut, artinya tidak ada degradasi selulosa menjadi gula terlarut. Indeks sobek dari kertas yang dihasilkan melalui perlakuan dengan CBD memperlihatkan tidak ada perubahan yang signifikan. Nilai tensile index, burst index, ring crush dan Concora Medium Test (CMT) dari kertas yang dihasilkan melalui perlakuan dengan CBD meningkat.Kata kunci: cellulose-binding domain, endoglukanase Egl-II, perbaikan freeness, kertas bekas

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Characterization of EngF from Clostridium cellulovorans and Identification of a Novel Cellulose Binding Domain

Applied and Environmental Microbiology ◽

10.1128/aem.64.3.1086-1090.1998 ◽

1998 ◽

Vol 64 (3) ◽

pp. 1086-1090 ◽

Cited By ~ 14

Author(s):

Akihiko Ichi-ishi ◽

Salah Sheweita ◽

Roy H. Doi

Keyword(s):

Amino Acids ◽

Cellulose Degradation ◽

Binding Activity ◽

Binding Domain ◽

Cellulose Binding Domain ◽

Binding Studies ◽

Endoglucanase Activity ◽

Clostridium Cellulovorans ◽

C Terminus ◽

Cellulose Binding

ABSTRACT The physical and enzymatic properties of noncellulosomal endoglucanase F (EngF) from Clostridium cellulovorans were studied. Binding studies revealed that the Kd and the maximum amount of protein bound for acid-swollen cellulose were 1.8 μM and 7.1 μmol/g of cellulose, respectively. The presence of cellobiose but not glucose or maltose could dissociate EngF from cellulose. N- and C-terminally truncated enzymes showed that binding activity was located at some site between amino acid residues 356 and 557 and that enzyme activity was still present when 20 amino acids but not 45 amino acids were removed from the N terminus and when 32 amino acids were removed from the C terminus; when 57 amino acids were removed from the C terminus, all activity was lost. EngF showed low endoglucanase activity and could hydrolyze cellotetraose and cellopentaose but not cellotriose. Activity studies suggested that EngF plays a role as an endoglucanase during cellulose degradation. Comparative sequence analyses indicated strongly that the cellulose binding domain (CBD) is different from previously reported CBDs.

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THE SEPARATION OF CELLULOSE-BINDING DOMAIN FROM ENDOGLUCANASE EGL-II BY PROTEOLYSIS METHOD

JURNAL SELULOSA ◽

10.25269/jsel.v3i01.42 ◽

2016 ◽

Vol 3 (01) ◽

Author(s):

Rina Masriani ◽

Taufan Hidayat ◽

Dewi Christanti Trisulo

Keyword(s):

Molecular Weight ◽

Protein Degradation ◽

Catalytic Domain ◽

Protein Molecule ◽

Binding Domain ◽

Intact Protein ◽

Crystalline Cellulose ◽

Cellulose Binding Domain ◽

Paper Fibers ◽

Cellulose Binding

Protein molecule of endoglucanase Egl-II is consisted of two domains, namely cellulose-binding domain (CBD) which serves in promoting the adsorption of the enzyme to the insoluble crystalline cellulose and cellulase catalytic domain which is responsible for the hydrolysis reaction. In this study, CBD of endoglucanase Egl-II was separated from the intact protein by degradation using the papain and then separation by ultrafiltration methods. The CBD resulted can be used to modify the waste paper fibers. The results of electrophoresis before degradation showed that endoglucanase Egl-II has a molecular weight about 57.5 kDa. The electrophoregram after protein degradation and separation of CBD from the endoglucanase Egl-II showed that CBD was separated from the intact protein with a molecular weight about 21 kDa. The yield of CBDs were 59.51%. It can be concluded that CBD of endoglucanase Egl-II can be separated from the intact protein.Keywords: endoglucanase Egl-II, cellulose-binding domain, papain, ultrafiltration. ABSTRAKMolekul protein dari endoglukanase Egl-II terdiri dari dua domain, yaitu cellulose-binding domain (CBD) yang berfungsi untuk mempromosikan adsorpsi enzim ke selulosa kristalin dan domain katalitik yang bertanggung jawab dalam reaksi hidrolisis. Dalam studi ini, CBD dari endoglukanase Egl-II telah dipisahkan dari protein utuhnya setelah didegradasi dengan papain. Pemisahan CBD dari campuran hasil degradasi dilakukan dengan metode ultrafiltrasi. CBD yang dihasilkan dapat digunakan untuk memodifikasi serat kertas bekas. Hasil elektroforesis sebelum degradasi menunjukkan endoglukanase Egl-II memiliki massa molekul sekitar 57,5 kDa. Dari elektroforegram setelah degradasi protein dan pemisahan CBD dari endoglukanase Egl-II memperlihatkan bahwa CBD telah terpisah dari protein utuhnya dan memiliki massa molekul sekitar 21 kDa. Rendemen CBD adalah 59,51%. CBD dari endoglukanase Egl-II dapat dipisahkan dengan metode ini.Kata kunci: endoglukanase Egl-II, cellulose-binding domain, papain, ultrafiltrasi.

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Cellulose binding domain from Clostridium cellulovorans as a paper modification reagent

Nordic Pulp & Paper Research Journal ◽

10.3183/npprj-2003-18-04-p421-428 ◽

2003 ◽

Vol 18 (4) ◽

pp. 421-428 ◽

Cited By ~ 3

Author(s):

llan Levy ◽

Tzur Paldi ◽

Oded Shoseyov ◽

Dan Siegel

Keyword(s):

Binding Domain ◽

Cellulose Binding Domain ◽

Clostridium Cellulovorans ◽

Cellulose Binding

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Cellulose as an inert matrix for presenting cytokines to target cells: production and properties of a stem cell factor–cellulose-binding domain fusion protein

Biochemical Journal ◽

10.1042/bj3390429 ◽

1999 ◽

Vol 339 (2) ◽

pp. 429-434

Author(s):

J. Greg DOHENY ◽

Eric J. JERVIS ◽

M. Marta GUARNA ◽

R. Keith HUMPHRIES ◽

R. Antony J. WARREN ◽

...

Keyword(s):

Stem Cell ◽

Fusion Protein ◽

Stem Cell Factor ◽

Direct Analysis ◽

Binding Domain ◽

Target Cells ◽

Cellulose Binding Domain ◽

Binding Characteristics ◽

Cellulose Binding ◽

Cellulose Surface

A chimaera of stem cell factor (SCF) and a cellulose-binding domain from the xylanase Cex (CBDCex) effectively immobilizes SCF on a cellulose surface. The fusion protein retains both the cytokine properties of SCF and the cellulose-binding characteristics of CBDCex. When adsorbed on cellulose, SCF–CBDCex is up to 7-fold more potent than soluble SCF–CBDCex and than native SCF at stimulating the proliferation of factor-dependent cell lines. When cells are incubated with cellulose-bound SCF–CBDCex, activated receptors and SCF–CBDCex co-localize on the cellulose matrix. The strong binding of SCF–CBDCex to the cellulose surface permits the effective and localized stimulation of target cells; this is potentially significant for long-term perfusion culturing of factor-dependent cells. It also permits the direct analysis of the effects of surface-bound cytokines on target cells.

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Application of cellulosic materials as supports for single-step purification and immobilization of a recombinant β-galactosidase via cellulose-binding domain

International Journal of Biological Macromolecules ◽

10.1016/j.ijbiomac.2022.01.006 ◽

2022 ◽

Author(s):

Adriano Gennari ◽

Renate Simon ◽

Nathalia Denise de Moura Sperotto ◽

Cristiano Valim Bizarro ◽

Luiz Augusto Basso ◽

...

Keyword(s):

Single Step ◽

Binding Domain ◽

Cellulose Binding Domain ◽

Cellulosic Materials ◽

Single Step Purification ◽

Cellulose Binding

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Optimization of chemical and enzymatic deinking of photocopier waste paper

BioResources ◽

10.15376/biores.6.1.447-463 ◽

2010 ◽

Vol 6 (1) ◽

pp. 447-463 ◽

Cited By ~ 9

Author(s):

Puneet Pathak ◽

Nishi K. Bhardwaj ◽

Ajay K. Singh

Keyword(s):

Physical Properties ◽

Removal Efficiency ◽

Strength Properties ◽

Waste Paper ◽

Cellulase Enzyme ◽

Tensile Index ◽

Drainage Time ◽

Tear Index ◽

The World ◽

Better Than

The utilization of post-consumer papers in the production of new paper products is increasing all over the world in recent years. Recycling of photocopier paper is a major problem due to difficulty in removal of non-impact ink. Enzymes offer potential advantages in ecofriendly deinking of recovered paper. In this study the deinking of photocopier paper was examined using chemicals and a commercial cellulase enzyme. Parameters of deinking experiments were optimized for hydrapulping. The ink was removed by flotation and washing processes. Then these parameters were compared in terms of ink removal ability of the process, as well as optical and strength properties of the deinked paper. The application of enzymatic deinking improved ink removal efficiency by 24.6% and freeness by 21.6% with a reduction in drainage time of 11.5% in comparison to those obtained with chemical deinking. The physical properties, namely burst index and tensile index, were observed to improve by 15.3% and 2.7%, respectively and brightness and tear index decreased by 2.1% and 21.9%, respectively. Results of deinking efficiency of photocopier paper showed that the enzyme used in the present work performed better than the conventional chemicals used for deinking.

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