• Asian-Australasian Journal of Animal Sciences
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• v.26 no.2
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• pp.253-259
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• 2013

Paddy rice is rarely used as a feed because of its high fiber content. In this study, two experiments were conducted to study the effects of supplementing an enzyme complex consisting of xylanase, beta-glucanase and cellulase, to paddy-based diets on the performance and nutrient digestibility in meat-type ducks. In the both experiments, meat-type ducks (Cherry Valley) were randomly assigned to four treatments. Treatment 1 was a basal diet of corn-soybean; treatment 2 was a basal diet of corn-paddy-soybean; treatment 3, had enzyme complex added to the corn-paddy-soybean basal diet at levels of 0.5 g/kg diet; and treatment 4, had enzyme complex added to the corn-paddy-soybean diet at levels of 1.0 g/kg diet. The results showed that the enzyme complex increased the ADG, and decreased the ADFI and F/G significantly (p<0.05) in the ducks, and the ADFI for the ducks fed the corn-paddy-soybean diet showed no difference compared to the ducks fed corn-soybean diets at all stages of the experiment (p<0.05). When corn was partially replaced by paddy, the digestibility of CP and NDF was decreased and increased, respectively (p<0.05), and the level of enzyme complex had a significant effect on both CP and NDF digestibility (p<0.05). As for the AME, addition of enzyme complex increased it significantly (p<0.05), but both diet types and levels of enzyme complex had no effect (p>0.05). The outcome of this research indicates that the application of enzyme complex made up of xylanase, beta-glucanase, and cellulase, in the corn-paddy-soybean diet, can improve performance and nutrition digestibility in meat-type ducks.

• Microbiology and Biotechnology Letters
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• v.18 no.6
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• pp.633-639
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• 1990

The genes for cellulases of Pseudomonas sp. LBC505 and CYC10, potent cellulase complex-producing strains, were cloned in Escherichia coli with pUC19. Recombinant plasmids pLCl and pLC2 were isolated from transformants producing cellulase by Congo red staining, and their genes cloned were 0.7 kb and 4.6 kb HindIII fragments, respectively. The inserts of pLCl and pLC2 were hybridized to chromosomal DNAs digested with HindIII from Pseudomona~ sp. LBC505 and CYC10, respectively. Immunodiffusion assays revealed that pLC1-and pLC2-encoded cellulase showed similarity with that of host strains. About 24% of cellulase activity was observed in the extracellular fraction of E. coli carrying pLC1, and its activity was higher about 1.4 times than that of LBC505. The enzymatic properties of pLC1 and pLC2 encoded cellulase were the same as those of cellulase from host strains. HPLC analysis and substrate specificity showed that cellulases were the same as those of cellulase from host strains. HPLC analysis and substrate specificity showed that cellulases cloned were endocellulase.

• Applied Biological Chemistry
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• v.12
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• pp.99-105
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• 1969

The yield of cellulase derived from Trichoderma $(O_2-1)$ was remarkably varied with various concentration of ethanol and acetone in purification of the enzyme. In the purification with ethanol of ${\beta}-glucosidase$, the best result was obtained in the concentration of 60% and, of CMCase and of filter paper disintegrating enzyme 80%. And in the purification with acetone of ${\beta}-glucosidase$, filter paper disintegrating enzyme, and CMCase, in the concentration of 60%, 80%, and 90% respectively, was shown the best yield. The activities of crude Cellulase preparation could be seperated into few of fractions by column chromatography with Silica gel, Cellulose powder, and gauze. Most of CMCase, avicelase, and ${\beta}-glucosidase$ were eluted, but most of filter paper disintegrating enzyme and the rest of enzymes mentioned the above were absorbed, and were eluted with water. Therefore, it was considered that CMCase is different from filter paper disintegrating enzyme in properties. The relative activity of CMCase was different from that of avicelase in the peak of elusion part. And it was considered that filter paper disintegrating enzyme and cellulose powder saccharifying enzyme was seperated respectively as absorption part and non absorption part. The auther came to the conclusion that at least there were more than three sorts of cellulase in Trichoderma $(O_2-1)$ cellulase preparation.

• Microbiology and Biotechnology Letters
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• v.19 no.4
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• pp.331-336
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• 1991

In order to utilize natural cellulosic materials as a fermentative substrate, saccharification of a various kind of native cellulosic materials was performed by using cellulase from the isolated strain, Pseudomonas sp. LBC-505 which potently produced cellulase complex and xylanase. Cellulase complex production was repressed by the low concentration of glucose, induced by cellulosic compounds such as CMC, wheat bran and rice straw et al. and showed to be highest on the PY-CMC medium containing 5% (w/v) wheat bran instead of CMC. Optimal temperature for enzyme reactions of CMCase and xylanase was $50^{\circ}C$, and $55^{\circ}C$ for $\beta$-glucosidase. Optimal pH for these enzyme reaction was 6.6. Rate of saccharification for natural cellulose was low by the treatment of crude enzyme. Among their substrates, rice straw was the most effective substrate of enzymatic reaction in this work. After treating rice straw with 5% (v/v) HC1 and hydrolysing with crude enzyme, rate of saccharification was 18.4% (w/w) on dry substrate. Sugars of cellulosic hydrolyzate mainly contained glucose, xylose and cellobiose.

• Journal of Microbiology and Biotechnology
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• v.20 no.5
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• pp.893-903
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• 2010

A cellulolytic and xylanolytic enzyme complex-producing alkalothermoanaerobacterium strain, Tepidimicrobium xylanilyticum BT14, is described. The cell was Grampositive, rod-shaped, and endospore-forming. Based on 16S rRNA gene analysis and various lines of biochemical and physiological properties, the strain BT14 is a new member of the genus Tepidimicrobium. The strain BT14 cells had the ability to bind to Avicel, xylan, and corn hull. The pH and temperature optima for growth were 9.0 and $60^{\circ}C$, respectively. The strain BT14 was able to use a variety of carbon sources. When the bacterium was grown on corn hulls under an anaerobic condition, a cellulolytic and xylanolytic enzyme complex was produced. Crude enzyme containing cellulase and xylanase of the strain BT14 was active in broad ranges of pH and temperature. The optimum conditions for cellulase and xylanase activities were pH 8.0 and 9.0 at $60^{\circ}C$, respectively. The crude enzyme had the ability to bind to Avicel and xylan. The analysis of native-PAGE and native-zymograms indicated the cellulosebinding protein showing both cellulase and xylanase activities, whereas SDS-PAGE zymograms showed 4 bands of cellulases and 5 bands of xylanases. Evidence of a cohesinlike amino acid sequence seemed to indicate that the protein complex shared a direct relationship with the cellulosome of Clostridium thermocellum. The crude enzyme from the strain BT14 showed effective degradation of plant biomass. When grown on corn hulls at pH 9.0 and $60^{\circ}C$ under anaerobic conditions, the strain BT14 produced ethanol and acetate as the main fermentation products.

• Food Science and Preservation
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• v.21 no.3
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• pp.442-450
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• 2014

In this study, we isolated a cellulase-producing bacterium isolated from traditional Korean fermented soybean paste and investigated the effect of culture conditions on the production of cellulase. This bacterium, which was identified as Bacillus subtilis 4-1 through 16S rRNA gene sequence analysis, showed the highest cellulase activity when the cells were grown at $45^{\circ}C$ for 24 hours in the CMC medium supplemented with 1.0% of soluble starch and 0.1% yeast extract. The initial optimum pH of the medium was observed in the range of 5.0~9.0. The optimal pH and temperature for the production of cellulase from B. subtilis 4-1 were pH 9.0 and $60^{\circ}C$ respectively. In addition, the enzyme showed significant activity in the temperature range of $20{\sim}90^{\circ}C$, which indicates that B. subtilis 4-1 cellulase is an alkaline-resistance and thermo-stable enzyme. This enzyme showed higher activity with CMC as the substrate for endo-type cellulase than avicel or pNPG as the exo-type substrates for exo-type cellulase and ${\beta}$-glucosidase. These results suggest that the cellulase produced from B. subtilis 4-1 is a complex enzyme rather than a mono-enzyme.

• Journal of Animal Science and Technology
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• v.46 no.5
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• pp.763-772
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• 2004

The patterns of fungal growth and fiber digestion under the microscope, and tile productions of fibrolytic enzyme were studied in an in vitro culture with Neocallimastix frontalis SA when either filter paper or rice straw was provided as sole energy source. Under the microscopic observation, active zoospores attachment, sporangium development and complex rhizoidal system were founded on the surface and at the edge of filter paper. After 7 days of incubation, a reduced fiber mass, a decreased fiber cohesion and a weakened fiber structure by fungal digestion were clearly observed. Similar fungal development was observed with rice straw, but fungal growth and digestion took place mostly on the damaged and exposed portion of rice straw. Although there were some differences in absolute concentration and pattern, the concentration of both cellulase and xylanase increased with incubation time with the higher activity being obtained with filter paper. Their differences were large especially after 48 and 96hr of incubation(P< 0.05). The filter paper was more good inducer of cellulolytic and xylanolytic enzymes compared with complex substrate, rice straw. These findings suggest that the filter paper is the better energy source for N frontalis than the complex substrate, and structural disintegration by physical process is able to help rumen fungal growth on the lignified roughage although anaerobic rumen fungi have mechanical and enzymatic functions for fiber digestion.

• Journal of Microbiology and Biotechnology
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• v.16 no.8
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• pp.1269-1275
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• 2006

The selection of multienzyme complex-producing bacteria under aerobic condition was conducted for improving the degradation of lignocellulosic substances. The criteria for selection were cellulase and xylanase enzyme production, the presence of cellulose-binding domains and/or xylan-binding domains in enzymes to bind to insoluble substances, the adhesion of bacterial cells to insoluble substances, and the production of multiple cellulases and xylanases in a form of a high molecular weight complex. Among the six Bacillus strains, isolated from various sources and deposited in our laboratory, Paenibacillus curdlanolyticus B-6 strain was the best producer of cellulase and xylanase enzymes, which have both cellulose-binding factors (CBFs) and xylan-binding factors (XBFs). Moreover, multiple carboxymethyl cellulases (CMCases) and xylanases were produced by the strain B-6. The zymograms analysis showed at least 9 types of xylanases and 6 types of CMCases associated in a protein band of xylanase and cellulase with high molecular weight. These cells also enabled to adhere to both avicel and insoluble xylan, which were analyzed by scanning electron microscopy. The results indicated that the strain B-6 produced the multienzyme complex, which may be cellulosome or xylanosome. Thus, P. curdlanolyticus B-6 was selected to study the role and interaction between the enzymes and their substrates and the cooperation of multiple enzymes to enhance the hydrolysis due to the complex structure for efficient cellulases and xylanases degradation of insoluble polysaccharides.

• Journal of the Korean Wood Science and Technology
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• v.38 no.3
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• pp.251-261
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• 2010

Cellulase from Formiptosis pinicola KMJ812 is an efficient cellulose degradation enzyme complex, especially with a high ${\beta}$-glucosidase activity. In this study, the change in enzymatic characteristics by immobilization and the reduction of immobilized enzyme activity by repeated usages were evaluated using cellulases from F. pinicola KMJ812. Among tested four resins, Duolite A568 resin had the best enzyme activity yield with 61.7% cellulase activity and 64.4% ${\beta}$- glucosidase activity during the cellulase immobilization. The best reaction temperature was $55^{\circ}C$ for both cellulase and ${\beta}$-glucosidase activities which were higher than the unimmobilized soluble cellulases. The best reaction pH was 4.0 for cellulase activity which was a little more basic than a soluble form and 4.5 for ${\beta}$-glucosidase activity. The immobilized cellulase activity was remained 98% of the beginning activity after 72 h incubation at $50^{\circ}C$ and 50% of the beginning activity after eight times usage at $50^{\circ}C$.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1986.12a
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• pp.524.1-524
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• 1986

CMCase, a member of cellulose decomposing enzymes, hydrolyze cellulose up to cellobiose. Cellobiase splits cellobiose to glucose units. Therefore, a linkage of the twogenes coding for CMCase and cellobiase on the same plasmid is needed to produce a cellulase complex which can produce glucose from cellulose. A genetic operon in which the two structural genes are under the control of a single promoter would be ideal for this purpose. The present report is on the linking of the two cellulase genes in one plasmid as a preliminary step of the operon construction.