• Journal of Industrial Technology
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• v.29 no.B
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• pp.121-125
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• 2009

New cellulase genes, named as CelV2 and CelN1, respectively, were isolated from Erwinia carotovora ATCC15713 and expressed in E. coli. The CelV2 and CelN1 gene were PCR amplified with degenerated primers and PCR products were sequenced and expressed in E. coli. Two new cellulase genes showed 97% homologies with previously reported Erwinia cellulase genes. The recombinant cellulase were purified with Ni-NTA column chromatography and its enzymatic properties were characterized. The optimum temperature of two enzymes were about $50^{\circ}C$ degree and optimum pH were around pH7.0. The newly isolated celluase genes could be used for enhancing substrate range of alcohol-producing bacteria such as Zymomonas mobilis.

• Textile Coloration and Finishing
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• v.13 no.1
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• pp.9-17
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• 2001

The purpose of this study is to present basic data for the enzymatic modification of acetate fabrics. The weight loss and rate of weight loss of acetate fabrics increased with increasing NaOH concentration and treating time. Acetyl value decreased as the weight loss became higher. The weight loss of alkaline-treated acetate fabrics were directly proportional to the concentration and treating time of cellulase. The optimum temperature and pH in cellulase treatment were $55^\circ{C}$ and pH 3.5. The surface shape revealed that density of fiber decreased by alkaline-treatment. With the treating time of cellulase, fibrillation occurred. In case of higher weight loss in alkaline treatment, fibril is removed after 180 min. The tensile strength decreased by alkaline and cellulase treatment. Especially, in case of higher weight loss of alkaline treatment, tensile strength decreased suddenly. Alkaline treatment increased the drapability of acetates, while cellulase treatment increased it initially but decreased gradually with treatment time. The dyeability after alkaline treatment was improved for reactive dye, but deteriorated for disperse dye. The cellulase treatment of acetate lowered the dyeability for both types of dyes.

• Journal of the Korean Wood Science and Technology
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• v.44 no.3
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• pp.381-388
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• 2016

Biopolishing using cellulases was introduced in the production of cotton fabric in order to improve the quality of fabric environmental friendly and is commonly used in the textile industry. In this study, the application of a crude cellulase from Acanthophysium sp. KMF001, which was excellent for the saccharification of cellulose, on biopolishing was evaluated. The optimum treatment biopolishing condition was at $50^{\circ}C$ and pH 4.5 for 60 minutes with 10% crude cellulase of fabric weight. After the optimized biopolishing, the crude cellulase of Acanthophysium sp. KMF001 reduced the tensile strength of the tested cotton fabric less than a commercial cellulase. The appearance of the cotton fabric after the treatment of the crude cellulase of Acanthophysium sp. KMF001 was similar to the fabric after a commercial cellulase treatment. All these results support that the crude cellulase of Acanthophysium sp. KMF001 was a good biopolishing cellulase.

• Korean Journal of Microbiology
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• v.44 no.1
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• pp.69-73
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• 2008

Fermented soybeans contain microorganisms, diverse enzymes, and bioactive compounds. Few studies on cellulase in Chungkookjang exist. Oligosaccharides play diverse roles of bioactivity. Through Congo red test and activity staining, it was confirmed that the enzyme solution contained cellulase. Optimal pH and temperature of the cellulase produced by Bacillus licheniformis B1 were 10 and $40^{\circ}C$, respectively. Through TLC analysis, it was demonstrated that the enzyme solution degraded carboxymethyl cellulose (CMC), whose main products contained dimer and trimer oligosaccharides. Cellulase activity of barley-Chungkookjnag fermented by the strain increased, compared with that of Chungkookjang. The cellulase was found to be a ${\beta}$-1,4-glucanase through the analysis of the cloned gene, showing polymorphism at 32 amino acid sites in the coding range of amino acid 10 and 460.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2003.10b
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• pp.20-20
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• 2003

Isolation and culture of leaf protoplasts from two chilli cultivars (Capsicum annuum var. accumnatum and Bird chilli) were developed to enhance selection process in the somatic hybridization programmes. In order to isolate the protoplasts from leaves of these two chilli cultivars different incubation periods (3, 5 and 10 hours) were tested with combinations of enzyme mixtures containing cellulase and macerozyme. Leaves were incubated on three enzyme mixtures (2% cellulase +0.4% macerozyme, 1% cellulase +0.2% macerozyme and 0.5% cellulase +0.1% macerozyme in 13% mannitol) at 251oC in the dark. Three hours of incubation using 2% cellulase and 0.4% macerozyme was the best for the protoplast isolation of both chilli cultivars tested. The yield was 5${\times}$108protoplasts/m1/g leaf tissue in both chilli varieties. It was found that in the mixed nurse method using Nagata and Takebe (NT) medium supplemented with 1.0mg/12,4-D, NAA and BAP with 0.5M mannitol and 1.2% Sea Plaque agarose is the best medium for protoplast culture. Protoplasts of Capsicum annum var. accumnatum were alive for 14 days forming cell walls and initiating cell division.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2003.10a
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• pp.50-58
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• 2003

Isolation and culture of leaf protoplasts from two chilli cultivars (Capsicum annuum var. accumnatum and Bird chilli) were developed to enhance selection process in the somatic hybridization programmes. In order to isolate the protoplasts from leaves of these two chilli cultivars different incubation periods (3, 5 and 10 hours) were tested with combinations of enzyme mixtures containing cellulase and macerozyme. Leaves were incubated on three enzyme mixtures (2% cellulase + 0.4% macerozyme, 1% cellulase + 0.2% macerozyme and 0.5% cellulase + 0.1 % macerozyme in 13% mannitol) at 251oC in the dark. Three hours of incubation using 2% cellulase and 0.4% macerozyme was the best for the protoplast isolation of both chilli cultivars tested. The yield was 5 ${\times}$ 108protoplasts/ml/ g leaf tissue in both chilli varieties. It was found that in the mixed nurse method using Nagata and Takebe (NT) medium supplemented with 1.0mg/12,4-D, NAA and BAP with 0.5M mannitol and 1.2% Sea Plaque agarose is the best medium for protoplast culture. Protoplasts of Capsicum annum var. accumnatum were alive for 14 days forming cell walls and initiating cell division.

• Applied Biological Chemistry
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• v.14 no.1
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• pp.59-97
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• 1971

As a study on the cellulase of Myriococcum albomyces the culture media for enzyme formation and properties of its crude preparation were investigated and the crude enzyme preparation was further fractionated. The results are summarized as follows: 1. Wheat bran solid culture produced stronger activities of cellulase than rice bran or defatted soy bean meal solid culture. 2. Shaking culture with wheat bran, rice bran or defatted soy bean meal produced higher cellulase activities than solid culture with the corresponding media. 3. The enzyme formation was higher at $45^{\circ}C$ than at $37^{\circ}C$ or $50^{\circ}C$ regardless of the kind of culture medium. 4. The formation of CMCase activity was more promoted by organic nitrogen source than inorganic nitrogen source. 5. The formation of cellulase activities were increased 1.5 to 3.0-fold by adding CMC, Avicel or cellulose powder as an inducer into 5% wheat bran basal medium. 6. Cellulase production using a tank culture procedure with addition of CMC or Avicel as an inducer was the highest at fifth day and thereafter decreased slightly. 7. The crude enzyme preparation showed pH optimum in 4.0 to 4.5, and pH stability in the range of 3.5 to 8.0. Optimum temperature for the activity was $65^{\circ}C$ which was higher than among other cellulases and it was stable at $60^{\circ}C$ for 120 minutes. 8. Dialyzed crude enzyme was activated by $Ca^{++}$ and $Mg^{++}$, but inhibited by $Hg^{++}$, $Cu^{++}$ and $Ag^{+}$. 9. Four different types of cellulase, i. e., fraction I, fraction II-a, fraction II-b, and fraction III were purified from the culture filtrate of Myriococcum albomyces through a sequence of ammonium sulfate fractionation, and elution chromatography on DEAE-Sephadex A-25, Amberlite CG-25 type 2 and hydroxyapatite columns. 10. These four cellulase fractions were showed to be homogenous by electrophoresis and ultracentrifugation and also gave a typical ultraviolet absorption spectrum of protein. 11. Four purified fraction showed different specificity toward substrates, fraction I has a stronger activity toward Avicel, cellulose powder, and gauze than that of other cellulase fractions. Fraction II-a had a powerful activity toward cellobiose but it was almost inactive agaisnt fibrous cellulose contrary to fraction I. On the contrary, the main component fraction II-b had a fairly higher activity on CMC and Avicel. Activity of fraction II-b toward cellobiose was about one-third of that of fraction II-a and activity on Avicel was lower than that of fraction I. Fraction III had a more powerful activity in decreasing viscosity of CMC. 12. Final hydrolysis products of fibrous cellulose by each fraction were cellobiose and glucose. Whereas oligosaccharides were predominant in the early stage of hydrolysis, prolonged reaction produced more glucose than cellobiose. Fraction I and fraction II-a acted synergically on Avicel. 13. Optimum pH for the activities of cellulase fraction I, fraction II-a, fraction II-b and fraction III were found to be 5.5, 5.0, 4.0 and $4.0{\sim}4.5$, respectively. These fractions were found to be stable in the range of pH $3.0{\sim}7.5$. 14. Optimum temperature for the activities of fraction I, fraction II-a, fraction II-b, and fraction III were $50^{\circ}C$, $55^{\circ}C$, $60^{\circ}C$ and $55^{\circ}C$, respectively. No less of activity was found by heating 120 minutes at $55^{\circ}C$ and fraction II-a was more stable than the others at $60^{\circ}C$. 15. Fraction I and fraction II-b were activated by $Ca^{++}$ and $Mg^{++}$ but inhibited by $Hg^{++}$ and $Ag^{+}$.

• Journal of the Korean Society of Clothing and Textiles
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• v.22 no.8
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• pp.1032-1042
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• 1998

The purpose of this study is to measure, compare, and investigate the physical properties of rayon/cotton blend fabrics treated with cellulase and cellulase & specific degeneration silicon and to present basic data which is in development a value-added fabric. The results are as follows. The condition for the treatment of cellulase was at 55$^{\circ}C$, pH 4 and the weight loss increased as the concentration of cellulase and the treated time increased. On treatment with cellulase, the crossectional view & longitudinal view of fiber noticed remarkable crack as weight loss increased and tensile strength and elongation decreased, and pilling was enhanced remarkably. KOSHI was increased, NUMERI and FUKURAMI were decreased as weight loss increased. In the basic characteristic value of clothing wearning, shape stability and drapability were decreased, but air content was improved. On treatment with cellulase & silicon, the degree of damage in the crossectional view & longitudinal view of the fiber reduced. Tensile strength, elongation, moisture regain improved. KOSHI and FUKURAMI were reduced, NUMERI was improved rather than when it was treated with cellulase. Therefore handle was improved. In the basic characteristic value of the clothing wearing, shape stability, air content, drapability were improved.

• Applied Biological Chemistry
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• v.24 no.3
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• pp.186-193
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• 1981

Three fractions of carboxymethyl-cellulase (F-I, F-II, and F-III) and ${\beta}-glucosidase$ form Aspergillus niger were partially purified by ammonium sulfate fractionation. Sephadex G-150 and DEAE-Sephadex column chromatography. The optimum conditions such as pH and temperature and thermal inactivation properties of the enzymes were investigated. Arrhenius plots of F-II and F-III appeared as straight lines, whereas that of F-I was biphasic. The Z-values of F-II and F-III were $8^{\circ}C$ and $10^{\circ}C$ respectively, while that of F-I was $4^{\circ}C$ over $60{\sim}70^{\circ}C$ and $383^{\circ}C$ over $70{\sim}98^{\circ}C$. Three fractions and the crude extract of carboxymethyl-cellulase exhibited a similar optimum pH 4.3 and temperature of $60^{\circ}C$, while Z-value of crude extract $(21.5^{\circ}C)$ was much higher than that of the purified enzyme. Maximum activity of both purified and crude extract of ${\beta}-glucosidase$ was shown at pH 4.7 and $60^{\circ}C$, and z-value of the enzyme was $7^{\circ}C$.

• Korean Journal of Microbiology
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• v.14 no.3
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• pp.117-127
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• 1976

A cellulase fraction (F IV-1) purified to about 8-folds was obtained from crude cellulase prepared from the wheat bran culture of S.atra. The partial purification of the enzyme was made by DEAE Sephadex and Sephdex cloumn chromatography in conjuction with ammonium sulfate precipitation. After stading at various pH's for 22 hours at $20^{\circ}C$, F IV-1 was most stable at pH 5.0 but when the enzyme fraction was stood for 74 hours, the point of pH stability was raised to around pH 6.0-7.0. After heating at various temperatures for 1 hour, F IV-1 was most stable at $20^{\circ}C$. The optimal enzyme activities of F IV-1 were seen at pH 6.0 and $50^{\circ}C$. The optimal concentrations of $Zn^{++}\;and\;Ca^{++}$ for the activities of crude cellulase were 6 and 4 mM respectively, but $Ca^{++}$ inhibited the enzyme activity at concentrations below 2 mM and above 6mM. Both $Cu^{++}\;and\;Mn^{++}$ ions inhibited cellulase activities and a ocmplete inactivation of crude cellulase was achieved at concentratioins of 5 and 2 mM of ions respectively. When Na-CMC was used as substrate, the Km values of crude cellulase and F IV-1 were calculated to be $5{\times}10^{-4}\;and\;2{\times}10^{-5}mM$, and V values 32 and 1.35 mmoles/hour, respectively. The Ki values of $Mn^{++}$ for crude cellulase and F IV-1 were found to be $8{\times}10^{-2}\;and\;3{\times}10^{-2}\;mM\;while\;those\;of\;Cu^{++}\;were\;at\;2{\times}10^{-1}\;and\;1{\times}10^{-1}\;mM\;respectively.\;Both\;Mn^{++}\;and\;Cu^{++}$ showed competitive inhibition with substrate.