• Title/Summary/Keyword: cellulase C

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Erwinia carotovora 유래의 cellulase 유전자의 클로닝 및 대장균에서의 발현 (Cloning and expression of cellulase genes from Erwinia carotovora in E. coli)

  • 김세돈;최신건
    • 산업기술연구
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    • 제29권B호
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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.

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알칼리와 셀룰라아제 처리에 의한 아세테이트 직물의 표면 형태 및 성능의 변화 (Changes in Surface Shape and Physical Properties of Acetate Fabrics by Alkaline and Cellulase Treatment)

  • 이애진;이혜자;유혜자
    • 한국염색가공학회지
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    • 제13권1호
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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.

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목재부후균, Acanthophysium sp. KMF001, 유래 섬유소분해효소를 이용한 섬유 개량 (Biopolishing of Cotton Fabric using Crude Cellulases from Acanthophysium sp. KMF001)

  • 신금;윤새민;김주혜;김영균;김태종;김영숙
    • Journal of the Korean Wood Science and Technology
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    • 제44권3호
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    • pp.381-388
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    • 2016
  • 면직물 표면의 잔털을 제거하여 새 옷 같은 느낌을 주기 위한 환경친화적 방법으로 셀룰라아제를 이용한 바이오폴리싱 방법이 도입되어 산업적으로 널리 이용되고 있다. 본 연구에서는 목재의 당화능력이 우수한 Acanthophysium sp. KMF001 효소를 분리해내어 이 효소의 바이오폴리싱 효과를 평가하였다. 실험을 통해 선정한 최적 바이오폴리싱 처리조건은 $50^{\circ}C$, pH 4.5, 처리효소농도는 면직물 중량 대비 10%, 반응시간은 60분으로 나타났다. 실험결과 최적화된 조건에서 처리한 면직물의 인장강도는 감소율이 상업용 효소보다 적었으며 전자현미경 관찰결과 잔털제거효과가 우수하여 바이오폴리싱 효과가 있는 것으로 평가되었다. 따라서 실험에 사용한 Acanthophysium sp. KMF001이 바이오폴리싱에 효과적으로 적용 가능한 효소로 판단된다.

청국장 발효균주 Bacillus licheniformis B1의 ${\beta}$-1,4-glucanase 특성 (Characterization of ${\beta}$-1,4-Glucanase Activity of Bacillus licheniformis B1 in Chungkookjang)

  • 황재성;유형재;김성조;김한복
    • 미생물학회지
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    • 제44권1호
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    • pp.69-73
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    • 2008
  • 대두가 발효된 청국장에는 미생물, 다양한 효소와 생리활성물질이 존재한다. 청국장의 탄수화물을 분해하는 cellulase에 대한 연구는 많지 않다. Oligosaccharide는다양한 생리활성을 지니고 있다. Congo red test 및 활성염색을 통해, 효소액이 cellulase를 포함하는 것을 확인했다. 청국장 발효 균주 Bacillus licheniformis B1이 분비하는 cellulase활성의 최적 pH와 온도는 각각 10과 $40^{\circ}C$이었다. TLC분석을 통해 효소액은 carboxymethyl cellulose (CMC)를 분해하여 2탄당 이상을 생성함을 확인하였다. 본 균주를 이용해서 제조한 보리 청국장에서는 효소의 활성이 증가되었다. 본 균주의 cellulase 유전자를 클로닝하여 분석한 결과, 본 효소는 ${\beta}$-1,4-glucanase였으며, 전체 coding 영역 $10{\sim}460$번째 아미노산 영역 중, 32군데서 polymorphism을 보였다.

Isolation and culture of protoplasts from leaf tissue of Capsicum annnum var. accumnatum Fingerh and C. frutescens L. [Syn. C. minmum Roxb.] (Bird chilli)

  • Lee, Kue-Jae;Lee, Wang-Hyu
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2003년도 제10차 국제학술회의 및 추계정기 학술발표회
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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.

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Isolation and culture of protoplasts from leaf tissue of Capsicum annnum var. accumnatum Fingerh and C. frutescensL. [Syn. C. minimum Roxb.] (Bird chilli)

  • Lee, Kue-Jae;Lee, Wang-Hyu
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2003년도 심포지엄
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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.

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Myriococcum albomyces가 생산하는 Cellulase에 관한 연구 (Studies on the Cellulase produced by Myriococcum of albomyces)

  • 정동효
    • Applied Biological Chemistry
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    • 제14권1호
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    • pp.59-97
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    • 1971
  • Myriococcum albomycesf가 생산하는 섬유소 분해효소군에 관한 연구로서 호소생산배지 및 조효소의 성질을 규명하고 몇 가지 효소군으로 정제한바 다음과 같은 결과를 얻었다. 1. 밀기울 고채해양의 각 효소 활성은 쌀겨고체배양 및 탈지대두박고체배양의 그것보다 강하였다. 2. 밀기울진탕, 쌀겨진탕배양 및 대두박진탕배양등은 상기의 고체배양보다 각 효소의 활성이 우수하였다. 3. $45^{\circ}C$에서 배양한 것이 배양기의 종류에 관계없이 $37^{\circ}C$$50^{\circ}C$에서 배양한 것보다 각 효소의 활성이 강하였다. 4. CMCase는 무기 질소원보다 유기 질소원을 첨가하므로서 더욱 생성이 촉진되었다. 5. 기본밀기울진탕배양기에 CMC, Avicel, 여지분말 등의 indncer를 첨가 하므로서 각 효소활성은 $1.5{\sim}3$배나 증가되었다. 6. CMC와 Avicel을 inducer로 하여 jar formentor에서 배양할 때 작 효소의 활성은 대개 5일째에 최고에 달하였다. 7. Cellulae 조효소의 최적 pH는 $4.0{\sim}4.5$, pH안정성은 $3.5{\sim}8.0$이였다. 그리고 최적온도는 $65^{\circ}C$ 부근으로 다른 사상균의 cellulase에 비하여 높으며 온도안정성도 $60^{\circ}C$에서 120분으로 거의 실활되지 않았다. 8. 조효소의 활성은 $Ca^{++}$, $Mg^{++}$으로 부활되며, $Hg^{++}$, $Cu^{++}$, $Ag^{+}$는 강한 저해체였다. 그리고 투석으로 약간 그 활성이 저하되었다. 9. 배양액을 여과하고 황산암모니움 분획, DEA-E-sephadex A-25, Amberlite CG-50 및 hydroxy-apatite column chromatography로 Avicel, CMC, 여지 분말에 대하여 활성이 다른 4개의 fraction을 분리 하고 이를 cellulase fraction I, fraction II-a, fraction II-b 및 fraction III라고 명명하였다. 10. 이들 4개의 fraction은 전기 영동, 초원심상 및 자외선흡수 등으로 보아서 단일의 단백질로 생각되었다. 11. Fraction I은 Avicelase활성이 강하고, fraction II-a는 cellobiase 활성이 강하였다. 그리고 fraction II는 CMCase 활성이 강하였으며, fraction III는 CMC 점도감소 활성이 강하였다. 12. 섬유소질을 각 fraction으로 가수분해한 최종산물은 cellobiose 및 glucose였다. 그리고 fraction I과 fraction II-a는 Avicel을 협동적으로 분해하였다. 13. Fraction I의 최적 pH 5.5, fraction II-a는 pH 5.0, fraction II-b는 pH 4.0, fractionIII는 pH $4.0{\sim}4.5$이며, 각 fraction의 pH 안정성은 pH $3.0{\sim}7.0$이였다. 14. Fraction I의 최적온도는 $50^{\circ}C$, fractionII-a는 $55{\sim}60^{\circ}C$, fraction II-b 는 $60^{\circ}C$, fraction III는 $55^{\circ}C$이며 각 fraction의 열안정성은 $55^{\circ}C$ 부근에서 120분으로 거의 실활되지 않고 fraction II-a는 $60^{\circ}C$에서도 특별히 안정하였다. 15. Fraction I과 fraction II-b 활성은 $Ag^{++}$, $Hg^{++}$에 의하여 저해되며 $Ca^{++}$, $Mg^{++}$으로는 부활되었다.

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Aspergillus niger가 생산(生産)하는 섬유소(纖維素) 분해효소(分解酵素)의 정제(精製) 및 특성(特性) (Purification and Characterization of Cellulolytic Enzymes from Aspergillus niger)

  • 박관화;오태광;신재두
    • Applied Biological Chemistry
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    • 제24권3호
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    • pp.186-193
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    • 1981
  • Aspergillus niger가 분비하는 섬유소분해효소(纖維素分解酵素)를 Sephadex G-150 column을 통과시켜서 ${\beta}-glucosidase$와 CM-cellulase을 분리(分離)하고 CM-cellulase는 다시 DEAE-Sephadex A-50 column을 통과시켜서 세개의 CM-cellulase F I, F II , F III를 분리하여 그 특성(特性)을 각기 조사 하였다. F I의 최적조건(最適條件)은 pH 4.0 및 $50^{\circ}C$였고 열불활성화특성(熱不活性化特性)은 Arrhenius plot에서 2개의 기울기가 다른 직선을 나타냈다. 즉 $60{\sim}70^{\circ}C$ 사이에서는 Z값이 $4^{\circ}C$ 이고 $70{\sim}98^{\circ}C$ 사이에서는 $383^{\circ}C$였다. F II는 pH 4.7 및 $50^{\circ}C$에서 최적조건(最適條件)을 보였고 Z값은 $8^{\circ}C$, F III는 pH 4.3 및 $60^{\circ}C$에서 높은 활성(活性)을 가졌고 Z값은 $10^{\circ}C$였다. 조(粗) CM-cellulase의 경우는 최적조건(最適條件)이 pH 4.3 및 $60^{\circ}C$로 F I , F II 및 F III와 비슷한 경향을 보였으나 Z값은 $21.5^{\circ}C$로 큰 차이(差異)를 보였다. 한펀 ${\beta}-glucosidase$는 정제효소(精製酵素) 및 조효소(粗酵素)가 다 같이 pH 4.7 및 $60^{\circ}C$에서 가장 높은 활성(活性)을 나타냈으며 Z값은 $7^{\circ}C$였다.

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셀룰라아제 처리시 실리콘 첨가에 따른 레이온/면 혼방직물의 물성변화 (The Change of the Physical Properties of Rayon/Cotton Blend Fabrics Treated with Cellulase by Addition of Silicon)

  • 이선화;송화순
    • 한국의류학회지
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    • 제22권8호
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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.

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Stachybotrys atra에서 추출한 섬유소분해효소에 관한 연구 II (STudies on the Cellulolytic Enzymes of Stachybotrys atra)

  • 김영민;김은수
    • 미생물학회지
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    • 제14권3호
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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.

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