• Asian-Australasian Journal of Animal Sciences
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• v.19 no.5
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• pp.684-689
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• 2006

The present study examined the effects of Tween 80 on the attachment and hydrolytic activity of a cellulase enzyme against ball-milled cellulose (BMC), using the whole component (native CBH I) and the catalysis module (core CBH I) of carbohydrolase I purified from Trichoderma viride (Meicelase, Meiji Seika, Tokyo, Japan). The effects were evaluated as protein concentrations in the supernatant after mixing enzyme and substrate with Tween 80 at room temperature. Tween 80 decreased the adsorption of native CBH I and core CBH I onto BMC (p<0.001) and increased the amount of reducing sugars released from BMC by native CBH I (p<0.001). However, Tween 80 did not enhance the hydrolytic activity of core CBH I. Observations using SEM revealed that Tween 80 caused cellulose filter paper to swell and enhanced surface cracks and filaments caused by native CBH I but not by core CBH I. These results suggested that Tween 80 decreases enzyme adsorption to its substrate but enhances enzymatic activity.

• Journal of Microbiology and Biotechnology
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• v.20 no.12
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• pp.1681-1688
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• 2010

A screening for cellobiohydrolase (CBH) activity was performed and Fomitopsis pinicola KMJ812 was selected for further characterization as it produced a high level of CBH activity. An extracellular CBH was purified to homogeneity by sequential chromatography of F. pinicola culture supernatants. The molecular mass of the F. pinicola CBH was determined to be 64 kDa by SDS-PAGE and by size-exclusion chromatography, indicating that the enzyme is a monomer. The F. pinicola CBH showed a $t_{1/2}$ value of 42 h at $70^{\circ}C$ and catalytic efficiency of $15.8mM^{-1}s^{-1}(k_{cat}/K_m)$ for p-nitrophenyl-${\beta}$-D-cellobioside, one of the highest levels seen for CBH-producing microorganisms. Its internal amino acid sequences showed a significant homology with hydrolases from glycoside hydrolase family 7. Although CBHs have been purified and characterized from other sources, the F. pinicola CBH is distinguished from other CBHs by its high catalytic efficiency and thermostability.

• Journal of Microbiology and Biotechnology
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• v.21 no.7
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• pp.711-718
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• 2011

A highly efficient cellobiohydrolase (CBH)-secreting basidiomycetous fungus, Agaricus arvensis KMJ623, was isolated and identified based on its morphological features and sequence analysis of internal transcribed spacer rDNA. An extracellular CBH was purified to homogeneity from A. arvencis culture supernatant using sequential chromatography. The relative molecular mass of A. arvencis CBH was determined to be 65 kDa by SDSPAGE and 130 kDa by size-exclusion chromatography, indicating that the enzyme is a dimer. A. arvencis CBH showed a catalytic efficiency ($k_{cat}/K_m$) of 31.8 $mM^{-1}\;s^{-1}$ for p-nitrophenyl-${\beta}$-D-cellobioside, the highest level seen for CBH-producing microorganisms. Its internal amino acid sequences showed significant homology with CBHs from glycoside hydrolase family 7. Although CBHs have been purified and characterized from other sources, A. arvencis CBH is distinguished from other CBHs by its high catalytic efficiency.

• KSBB Journal
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• v.22 no.1
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• pp.16-21
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• 2007

Although Energy demands of modern society increase rapidly, current energy would be exhausted shortly. Therefore development of bio-ethanol production process from cellulose containing materials was extremly demanded. Therefore development of highly functional cellulase is requisite for this purpose. In this study cellobio-hydrolase (CBH1) gene from Trichorderma reesei was used to increase cellulase activity by directed evolution and highly functional cellobio-hydrolase was obtained and characterized.

• Mycobiology
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• v.40 no.2
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• pp.107-110
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• 2012

Genes encoding the cellobiohydrolase enzyme (CBHI), designated as cbhI, were isolated from the basidiomycetes Auricularia fuscosuccinea, Pleurotus giganteus, P. eryngii, P. ostreatus, and P. sajor-caju. Initially, the fungal genomic DNA was extracted using a modified cetyltrimethyl ammonium bromide (CTAB) protocol and used as a DNA template. The cbhI genes were then amplified and cloned using the pGEM-T Easy Vector Systems. The sizes of these PCR amplicons were between 700~800 bp. The DNA sequences obtained were similar showing high identity to the cbhI gene family. These cbhI genes were partial consisting of three coding regions and two introns. The deduced amino acid sequences exhibited significant similarity to those of fungal CBHI enzymes belonging to glycosyl hydrolase family 7.

• Journal of Radiation Industry
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• v.8 no.2
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• pp.65-69
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• 2014

The cellobiohydrolase gene (cbhI), a key component of a cellulolytic system, of a mutant PfCM4 (Pleurotus florida), developed through gamma ray radiation mutagenesis, was isolated and cloned. The deduced amino acid sequence was closely related to the glycoside hydrolase family 7 (GH7). The molecular weight of the deduced amino acid sequence of cbhI gene was found to be 22.4 kDa. Though the percent identity was found to be much less (35.61%) between the wild type and mutant, the cellulolytic activity of PfCM4 was 17.24% higher than that of the wild type. This shows that the catalytic domain of the cbhI gene was conserved in the mutant PfCM4.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.468-472
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• 2003

Recently, genetic engineering techniques have been used to display various heterologous peptides and proteins (enzyme, antibody, antigen, receptor and fluorescence protein, etc.) on the yeast cell surface. Living cells displaying various enzymes on their surface could be used repeatedly as 'whole cell biocatalysts' like immobilized enzymes. We constructed a yeast based whole cell biocatalyst displaying T. reesei cellobiohydrolase I (CBH I ) on the cell surface and endowed the yeast-cells with the ability to degrade cellulose. By using a cell surface engineering system based on ${\alpha}-agglutinin,$ CBH I was displayed on the cell surface as a fusion protein containing the N-terminal leader peptide encoding a Gly-Ser linker and the $Xpress^{TM}$ epitope. Localization of the fusion protein on the cell surface was confirmed by confocal microscopy. In this study, we report on the genetic immobilization of T. reesei CBH I on the S. cerevisiae and hydrolytic activity of cell surface displayed CBH I.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.499-503
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• 2003

In the study, we have obtained modified cellulase with higher cellulose degradation activity by molecular evolution method. Cellobiohydrolase(CBH I ) gene of Trichorderma reerri has been used. Cellulase mutant 228-G2 was selected and the activity of cellulase mutant 228-G2 was increased by 300% compared to original CBH I The 17 among 1542bases were found to be modified with mutant 228-G2.

• Bulletin of the Korean Chemical Society
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• v.22 no.7
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• pp.716-720
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• 2001

To test the metal ion effect, hydrolysis experiments for two cellobiohydrolases (CBHⅠ and CBH Ⅱ) from Trichoderma reesei have been carried out in the presence of 10 mM metal ions, such as Cu++, Mn++, Ca++, Hg++, Ba++, Pb++, and Cd++. The addition of Mn++, Ba++, and Ca++(10 mM) during the hydrolysis of Avicel PH 101 caused an increase in the total reducing sugar (TRS) for CBH Ⅰ by 142, 135, and 114 percent, respectively. Those for CBH Ⅱ increased by 177, 175, and 115 percent, respectively. The Mn++ was the most stimulatory metal ion, whereas Hg++ was the most inhibitory metal ion. The adsorption experiments were performed to investigate how the influence of Mn++ and Hg++ on the hydrolysis is related to the adsorption of cellobiohydrolases on cellulose. The increase in TRS during hydrolysis by adding Mn++ caused an increase in adsorption affinity (Kad) and tightness (ΔHa). While, the decrease of TRS during hydrolysis by adding Hg++ caused a decrease in the adsorption affinity (Kad) and tightness (ΔHa). These results indicate the changes in the tightness and affinity of adsorption by adding metal ions play a crucial role in the degradation of the microcrystalline cellulose.

• KSBB Journal
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• v.8 no.1
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• pp.75-82
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• 1993

The inhibitory effect of cacao bean husk (CBH) extract on glucosyltransferase(GTasc) from Streptococcus mutans B13 was investigated. Water solube extract from CBH showeda sarong inhibitory effect (88-89%) on GTase from Streptococcus mutans Bl3. GTase inhibitors from sequential extraction by hot water or water-methanol had the strongest inhibition. Sources, fermentation, and types of solvents and fumigation processes did not influence the effect. These active compounds proved to be polyphones through acid hydrolytic analysis of the precipitates by ammonium sulfate or ethanol and proteinase K. It was also confirmed by additional column chromatography of Sephadex G-50, Sephadex LH-20 and DEAE-Sephdex A-50.