• 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.

• Journal of Life Science
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• v.21 no.10
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• pp.1481-1486
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• 2011

Spent mushroom substrate (SMS) is a by-product remained after a crop of mushrooms. About seven thermophilic strains were isolated from SMS (Flammulina velvtipes). Among them, one isolate, designated UJ03, showed the antifungal activity against Aspergillus flavus and Aspergillus ochraceous producing mycotoxin on PDA medium, potentially. The strain UJ03 produced cellulase and xylanase as extracellular hydrolases. The strain UJ03 was identified as a member of the genus Bacillus by biochemical characteristics using Bacillus ID kit and VITEK 2 system. Comparative 16S rDNA gene sequence analysis showed that strain UJ03 formed a distinct phylogenetic tree within the genus Bacillus and was most closely related to Bacillus amyloliquefaciens with sequence similarity of 98.9%. On the basis of its physiological properties, biochemical characteristics and phylogenetic distinctiveness, strain UJ03 was classified within the genus Bacillus, for which the name Bacillus sp. UJ03 is proposed. The antifungal compound from Bacillus sp. UJ03 was similar to lipopeptide iturin A of Bacillus sp.

• 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 Society of Food Science and Nutrition
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• v.46 no.9
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• pp.1114-1121
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• 2017

Bacillus strains not producing harmful components were isolated from Korean traditional soybean products. Extracellular enzyme activities (amylase, protease, cellulase, and xylanase) of isolated Bacillus strains were measured, and Bacillus strains with high protease activity were selected. The selected 15 strains were identified as Bacillus amyloliquefaciens (10), Bacillus methylotrophicus (1), Bacillus velezensis (1), and Bacillus subtilis (3). Among them, B. subtilis JBG17019, B. amyloliquefaciens JBD17076, and B. amyloliquefaciens JBD17109 showed antimicrobial activities against food-borne microorganisms. The production abilities of glutamate, glutamine, and poly-${\gamma}$-glutamic acid (${\gamma}$-PGA) of the selected Bacillus strains were measured to analyze fermentation characteristics related to glutamic acid metabolism. The factor for multivariate was analyzed by the principal components analysis (PCA) method between fermentation characteristics and ${\gamma}$-PGA production. The three principal components were classified according to the PCA method: PC1 [enzyme activity (amylase, cellulase, and xylanase)], PC2 (${\gamma}$-PGA), and PC3 (protease, glutamate, and glutamine). As a result, B. amyloliquefaciens JBD17076 and B. subtilis JBG17019 strains were evaluated as having excellent enzyme activity and ${\gamma}$-PGA production.

• Journal of The Korean Society of Grassland and Forage Science
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• v.33 no.1
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• pp.58-66
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• 2013

This study was conducted in order to develop slaughterhouse rumen content (SRC) as a potential feed additive. The moisture content of SRC can reach 80%, and therefore an appropriate dewatering process is required before it can be used. In this study, the effects of heating temperature, heating time, and filler content during the dewatering process on the activity of various enzymes in SRC were investigated. The Box-Behnken experimental design was employed, involving a total of 45 experimental runs, consisting of three variables (heating time, heating temperature, and filler content) with three levels per variable (12, 30 and 48 hr; 60, 75 and $90^{\circ}C$; 12, 22.5 and 33% for heating time, heating temperature, and filler content, respectively). For enzyme activities, xylanase, cellulase, and amylase were examined, and the results were subjected to an analysis of variance. Heating time, heating temperature and filler content had significant effects on the activity of each enzyme (p<0.05). Cellulase and amylase activities decreased (p<0.05) at elevated heating temperatures, whereas xylanase was reasonably stable around $90^{\circ}C$. The activities of all enzymes decreased (p<0.05) with increased heating time. Optimum filler contents for xylanase, cellulase, and amylase activities were 22.5, 12 and 33%, respectively. However, optimum conditions for all variables that simultaneously maximize the activity of all three enzymes could not be ascertained in this study. Nevertheless, the results from the current study can be useful as basic information for the development of SRC as a feed additive enriched with improved major enzymes for livestock feed digestion.

• Applied Biological Chemistry
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• v.24 no.1
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• pp.7-14
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• 1981

Among the Basidiomycetes, Pleurotus ostreatus 301 and Lentinus edodes 3-1 were chosen because of their good enzyme productivity and rapid mycelial growth in rice straw medium. Their cultural conditions adequate for the enzymes production and effects of various materials and inorganic salts added to thd rice straw media were investigated. L. edodes 3-1 was excellent in productivity of cellulase and xylanase, and P. ostreatus 301 in protease. The optimum conditions for enzyme production were at $30^{\circ}C$ in cellulase production and at $25^{\circ}C$ in xylanase and protease production, with 75% moisture content and 5.0-6.0 initial pH. The appropriate cultural periods, for enzyme production were 30 days and 35 days for P. ostreatus 301 and L. edodes 3-1, respectively. Among the various materials added, defatted soybean, defatted rape seed, or defatted sesame were all effective to enzyme production but reduced mycelial growth. Rice bran was also effective, particularly at 30% concentration. The addition of inorganic salts showed effective to enzyme production. Among inorganic salts, optimum concentration of $CaCO_3$ was 5%, and that of $CaSO_4$ was 2%.

• Journal of Korea Technical Association of The Pulp and Paper Industry
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• v.37 no.1 s.109
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• pp.25-37
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• 2005

This study was carried out to examine the optimal cultural condition in enzyme activities of CMCase, FPase and xylanase in selected strains which secret extracellular enzymes for using deinking agent to old newsprint. The results of this study were as follow: The production of enzyme by Bacillus pumilus I was maximal as grown on the medium, containing of rice bran+xylan $2.0\%$, peptone $0.8\%,\;K_2HPO_4\;0.1\%\;and\;CaCl_2\;0.06\%$ at pH 8.0 and $28^{\circ}C$ for 72 hours. Optimal cultural condition of B. subtilis I was avicel+xylan $3.5\%,\;urea\;0.4\%,\;K_3PO_4\;0.1\%\;and\;CaCl_2\;0.015\%$ at pH 9.0 and $28^{\circ}C$ for 36 hours. The maximal enzyme production was observed in the medium, containing of avicel+xylan $3.5\%,\;urea\;1.6\%\;and\; K_2HPO_4\;0.125\%$ with pH 9.0 when B. pumilus II was cultured at $28^{\circ}C$ for 60 hours. The production of enzyme by B. subtilis IT was maximal as grown on the medium, containing of xylan $2.0\%,\;yeast\; extract\;0.6\%,\;K_2HPO_4\;0.1\%\;and\;ZnSO_4\;0.04\%$ at pH 8.0 and $34^{\circ}C$ for 36 hours. The activities of FPase and xylanase in tested 4 strains were not much different with Thermomonospora fusca.

• Journal of Microbiology and Biotechnology
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• v.13 no.1
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• pp.1-8
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• 2003

Strain FJI, a filamentous fungus isolated from rotten wood, showed high ability to hydrolyze cellulosic materials. To identify the strain FJI, ITS sequencing analysis and morphological observation were performed. The strain FJI was identified as Trichoderma harzianum. The strain produced a large amount of CMCase, xylanase, ${\beta}-glucosidase$, and avicelase. Optimal culture conditions for the production of the enzymes, such as pH, temperature, and inoculation concentration, were initial pH 6.0-7.0,$25-30^{\circ}C$, and $10^4$ ea-spores/ml in Mandel's medium, respectively. T.hanzianum FJI utilized various cellulosic materials and organic nitrogen sources to produce cellulases and xylanase, and also considerably a crystalline and/or insoluble material like Avicel and rice straw. The highest levels of CMCase and xylanase were 41.2 and 65.6 U/ml in 7 days of cultivation using 2.5% of carbon source (Avicel+CMC) and 0.5% of nitrogen source (peptone), respectively.

• Journal of Microbiology and Biotechnology
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• v.24 no.11
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• pp.1525-1535
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• 2014

The xynC gene, which encodes high molecular weight xylanase from Paenibacillus sp. DG-22, was cloned and expressed in Escherichia coli, and its nucleotide sequence was determined. The xynC gene comprised a 4,419bp open reading frame encoding 1,472 amino acid residues, including a 27 amino acid signal sequence. Sequence analysis indicated that XynC is a multidomain enzyme composed of two family 4_9 carbohydrate-binding modules (CBMs), a catalytic domain of family 10 glycosyl hydrolases, a family 9 CBM, and three S-layer homologous domains. Recombinant XynC was purified to homogeneity by heat treatment, followed by Avicel affinity chromatography. SDS-PAGE and zymogram analysis of the purified enzyme identified three active truncated xylanase species. Protein sequencing of these truncated proteins showed that all had identical N-terminal sequences. In the protein characterization, recombinant XynC exhibited optimal activity at pH 6.5 and $65^{\circ}C$ and remained stable at neutral to alkaline pH (pH 6.0-10.0). The xylanase activity of recombinant XynC was strongly inhibited by 1 mM $Cu^{2+}$ and $Hg^{2+}$, whereas it was noticeably enhanced by 10 mM dithiothreitol. The enzyme exhibited strong activity towards xylans, including beechwood xylan and arabinoxylan, whereas it showed no cellulase activity. The hydrolyzed product patterns of birchwood xylan and xylooligosaccharides by thin-layer chromatography confirmed XynC as an endoxylanase.

• 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.