• Food Science and Preservation
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• v.24 no.1
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• pp.60-67
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• 2017

In this study, we tried to establish the best method for fresh blueberry beverage production using enzyme treatment as well as low temperature extraction. During extraction of physiologically functional materials, we used low temperature to prevent nutritional loss by heat. In addition, we investigated optimal blueberry extraction conditions using various enzyme treatments (cellulase, pectinase, cellulase:pectinase (1:1) mixture) to increase extraction efficiency and reduce turbidity. A variety and ratio of enzymes, extraction temperature, extraction time, and shaking speed were considered for the best extraction efficiency rate. We observed high extraction efficiency rates of 85.72-86.55% and 87.06-87.93%, respectively, upon cellulase or pectinase treatment. In addition, a mixture of cellulase:pectinase (1:1) showed an extraction efficiency rate of 86.84-88.14%. The best extraction efficiency rate was observed when crude blueberry was treated at $45^{\circ}C$ (87.91%), for 3 h (87.88%), in a 90 rpm shaker (89.19%). Sugar content and acidity of blueberry extract were not affected by the various treatments. However, total phenolic compounds were detected upon pectinase treatment (18.62 mg/g). Only fructose and glucose as free sugars were found in all samples regardless of treatments and extraction conditions.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.2
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• pp.204-210
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• 2016

This study examined the effect of supplementing exogenous cellulase on nutrient and energy utilization. Twelve desexed Boer crossbred goats were used in a replicated $3{\times}3$ Latin square design with 23-d periods. Dietary treatments were basal diet (control, no cellulase), basal diet plus 2 g unitary cellulase/kg of total mixed ration dry matter (DM), and basal diet plus 2 g compound cellulase/kg of total mixed ration DM. Three stages of feeding trials were used corresponding to the three treatments, each comprised 23 d, with the first 14 d as the preliminary period and the following 9 d as formal trial period for metabolism trial. Total collection of feces and urine were conducted from the 4th d of the formal trial, and gas exchange measures were determined in indirect respiratory chambers in the last 3 d of the formal trial. Results showed that cellulase addition had no effect (p>0.05) on nutrient digestibility. Dietary supplementation of cellulase did not affect (p>0.05) N intake and retention in goats. Gross energy (GE) intake, fecal energy and urinary energy excretion, heat production were not affected (p>0.05) by the cellulase supplementation. Total methane emission (g/d), $CH_4$ emission as a proportion of live weight or feed intake (DM, organic matter [OM], digestible DM or digestible OM), or $CH_4$ energy output ($CH_4$-E) as a proportion of energy intake (GE, digestible energy, or metabolizable energy), were similar (p>0.05) among treatments. There was a significant (p<0.001) relationship between $CH_4$ and live weight (y = 0.645x+0.2, $R^2$ = 0.54), $CH_4$ and DM intake (y = 16.7x+1.4, $R^2$ = 0.51), $CH_4$ and OM intake (y = 18.8x+1.3, $R^2$ = 0.51) and $CH_4$-E and GE intake. Results from this study revealed that dietary supplementation of cellulase may have no effect on nutrient digestibility, nitrogen retention, energy metabolism, and methane emission in goat.

• Journal of Life Science
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• v.17 no.7 s.87
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• pp.983-989
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• 2007

Bacillus lichemiformis Kll, a plant growth promoting rhizobacterium was reported as a producer of auxin, siderophore, as well as antifungal cellulase under some culture conditions. In vitro test, B. licheniformis Kll represented excellent antagonistic ability against Fusarium oxyspoum (KACC 40037), and showed broad spectrum against other phytopathogenic fungi. B. licheniformis Kll had cellulolytic activity toward not only carboxymethyl-cellulose (CMC) but also insoluble cellulose, such as fungal cell wall cellulose, filter paper (Whatman No. 1), and Avicel. In addition, we confirmed antifungal substance production by butanol-extract methods. The strain produced optimally the antifungal substance when it was cultivated at pH 9.0, 30${\circ}$C for 4 days on nutrient medium. The biological control mechanisms of B. lichemiformis Kll were caused by antifungal substance, cellulase and siderophore against phytopathogenic fungi.

• Applied Biological Chemistry
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• v.19 no.3
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• pp.130-138
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• 1976

In order to utilize the agricultural waste products, two strains of mold producing powerful cellulolytic enzyme were sereened from various soils, composts, rotten wood and others. The optimum condition of cellulase production was studied. The results obtained were summarized as follows. 1. Two strains of mold which showed remarkably high cellulolytic activity were identified as Aspergillus niger-SM 6 and Trichoderma viride-SM 10. 2. The highest cellulase production was obtained at pH 5.0-6.0 in 5 days. 3. Cellulase production in strain Aspergillus niger-SM 6 increased with the addition of C.M.C., $(NH_4)_2SO_4$, C.S.L., orange peel powder and rice hull. The rice hull, treated with 3N NaOH at $120^{\circ}C$ for 15 min. and neutralized with various acids, was used. Up to 50% of wheat bran could be substituted by the treated rice hull without any decrease of cellulase activity. 4. In the strain of Trichoderma viride-SM 10, cellulase production increased with the addition of C.M.C., $NH_4NO_3$, Vitamin-free casamino acid and orange peel powder, while the other carbon, nitrogen, phosphate sources, natural nutrients and organic substances gave no remarkable effect.

• Asian-Australasian Journal of Animal Sciences
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• v.11 no.5
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• pp.522-529
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• 1998

This experiment was conducted to study the effects of lactic acid bacteria (LAB) inoculation either alone or in combination with cell wall degrading enzymes on the fermentation characteristics and chemical compositions of Rhodesgrass silage. Over to 1 kg of fresh Rhodesgrass sample a treatment of inoculant LAB with or without addition of an enzyme of Acremoniumcellulase (A) or Meicelase (M) or a mixture of both enzymes (AM) was applied. The treatments were control untreated, LAB-treated (application rate $1.0{\times}10^5cfu/g$ fresh sample), LAB+A 0.005%, LAB+A 0.01%, LAB+A 0.02%, LAB+M 0.005%, LAB+M 0.01%, LAB+M 0.02 %, LAB+AM 0.005%, LAB+AM 0.01%, and LAB+AM 0.02%. The sample was ensiled into 2-L vinyl bottle silo, with 9 silages of each treatment were made. Three silages of each treatment were incubated at 20, 30 and $40^{\circ}C$ for 2-months of storage period. All silages were well preserved with their fermentation quality has low pH values (3.91-4.26) and high lactic acid concentrations (4.11-9.89 %DM). No differences were found in fermentation quality and chemical composition of the control untreated silage as compared to the LAB-treated silage. Combined treatment of LAB+cellulases improved the fermentation quality of silages measured in terms of lower (p < 0.01) pH values and higher (p < 0.05) lactic concentrations than those of LAB-treated silages. Increasing amount of cellulase addition resulted in decrease (p < 0.05) of pH value and increase (p < 0.05) of lactic acid concentration. LAB + cellulase treatments (all cellulase types) reduced (p < 0.01) NDF, ADF and in vitro dry matter digestibility of silages compared with the control untreated silages. The fermentation quality and the rate of cell wall reduction were higher (p < 0.01) in the silages treated with LAB + cellulase A than in the silages treated with either LAB+cellulase M or LAB + cellulase AM. Incubation temperature of $40^{\circ}C$ was likely to be more appropriate environment for stimulating the fermentation of Rhodesgrass silages than those of 20 and $30^{\circ}C$.

• Microbiology and Biotechnology Letters
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• v.34 no.4
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• pp.311-317
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• 2006

Using PCR amplification, we cloned a cellulase gene (ce/H) from the Bacillus subtilis AH18 which has plant growth-promoting activity and antagonistic ability against pepper blight caused by Phytophthora capsici. The 1.6 kb PCR fragment contained the full sequence of the cellulase gene and the 1,582 bp gene deduced a 508 amino acid sequence. Similarity search in protein database revealed that the cellulase of B. subtilis AH18 was more than 98% homologous in the amino acid sequence to those of several major Bacillus spp. The ce/H was expressed in E. coli under an IPTG inducible lac promoter on the vector, had apparent molecular weight of about 55 kDa upon CMC-SDS-PAGE analysis. Partially purified cellulase had not only cellulolytic activity toward carboxymethyl-cellulose (CMC) but also insoluble cellulose, such as Avicel and filter paper (Whatman No. 1). In addition, the cellulase could degrade a fungal cell wall of Phytophthora capsici. The optimum pH and temperature of the ce/H coded cellulase were determined to be pH 5.0 and $50^{\circ}C$. The enzyme activity was activated by $AgNO_3$ or $CoCl_2$. However its activity was Inhibited by $HgC1_2$. The enzyme activity was activated by hydroxy urea or sodium azide and inhibited by CDTA or EDTA. The results indicate that the cellulase gene, ce/H is an antifungal mechanism of B. subtilis AH18 against phytophthora blight disease in red-pepper.

• 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.

• Microbiology and Biotechnology Letters
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• v.25 no.6
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• pp.546-551
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• 1997

A bacterium producing the extracellular cellulases was isolated from soil and has been identified as Bacillus sp. The isolate, named Bacillus sp. 79-23, was shown to be very similar to B. subtilis on the basis of its biochemical properties. The carboxymethyl cellulase (CMCase) of culture supernatant was most active at 60$\circ$C and pH 6.0, and retained 90% of its maximum activity at pH 7.0. The additional carbon sources affected the CMCase productivity than nitrogen sources in the culture medium. The carbon sources including wheat bran, rice straw, maltose and glucose increased the enzyme productivty. Especially, the maximum CMCase production was 5.2 units/ml in LB medium supplemented with 3% (w/v) wheat bran, which was 13-folds more than that in LB medium. It was found that the enzyme production was in association with the growth of Bacillius sp. 79-23. But, whean bran did not affect the growth of isolate, suggesting that increasement of CMCase production was owing to the induction of CMCase biosynthesis by wheat bran. In addition, both water-soluble and insoluble components of wheat bran was involved in induction of CMCase biosynthesis.

• Fashion & Textile Research Journal
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• v.11 no.3
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• pp.465-468
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• 2009

In this study, acid cellulase was used to treat denim fabrics by varying pH, temperature, enzyme concentration, treatment time and non-ionic surfactant (Triton X-100) concentration. Treatment condition was controlled based on the weight loss. The characteristics of enzyme-treated fabrics were measured in terms of tearing strength, stiffness, and color difference. The optimum conditions for cellulase treatment of denim fabric were pH 5.0, $50^{\circ}C$, 3% (o.w.f.), 90minutes. The weight loss did not change significantly with the addition of a non-ionic surfactant, but it improved when more non-ionic surfactant were used. The tearing strength of enzyme-treated denim fabrics did not deteriorate. The stiffness of the treated fabrics improved with the enzymatic treatment with and without the non-ionic surfactant. The difference in color of fabrics treated with enzyme increased.

• Preventive Nutrition and Food Science
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• v.20 no.3
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• pp.210-214
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• 2015

Grape pomace is an abundant source of underutilized winery by-products. Polyphenols were extracted from grape pomace using cellulase and gluco-amylase enzymes. 2,2-diphenyl-1-picrylhydrazyl (DPPH) and Folin-Ciocalteu's assays were used to measure antioxidant activity and total polyphenolic contents. Both cellulase, and gluco-amylase digested grape pomace showed efficient radical scavenging activity. In addition, the total polyphenolic contents of cellulase digested grape pomace showed lower concentrations were effective compared to higher concentrations, whereas glucoamylase enzyme did not show remarkable variations. The DPPH radical scavenging activity and total polyphenolic contents were significantly higher in the cellulase digested grape pomace compared to the gluco-amylase digested and the not digested grape pomace. It is notable that enzymatic digestions were efficient for extracting polyphenols from grape pomace. The underutilized grape pomace polyphenols can be further used for food safety as a natural antioxidant.