• 제목/요약/키워드: cellulase,

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반응 표면 분석법을 사용한 Bacillus subtilis NC1 유래 cellulase 생산 배지 최적화 (Optimization of a Medium for the Production of Cellulase by Bacillus subtilis NC1 Using Response Surface Methodology)

  • 양희종;박창수;양호연;정수지;정성엽;정도연;강대욱;문자영;최낙식
    • 생명과학회지
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    • 제25권6호
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    • pp.680-685
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    • 2015
  • 이전에 토양으로부터 cellulase와 xylanase 생산 균주로 단리하였다. 단리한 균주 유래의 16S rRNA 유전자 및 API 50 kit를 분석한 결과 Bacillus subtilis와 약 99.5%의 높은 상동성을 보였기에 본 균주를 B. subtilis NC1으로 명명하였다. Bacillus subtilis NC1 균주 유래 cellulase와 xylanase 유전자를 cloning 하여 유전자 배열을 규명하였다. 또한, 두 효소의 아미노산 배열을 이용하여 상동성을 검토한 결과 cellulase는 Glycoside hydrolase family (GH) 5 그리고 xylanase는 GH30에 속하는 효소임을 밝혔다. 본 연구에서는 B. subtilis NC1 의 cellulase 생산을 위한 배지성분의 최적 농도를 결정하기 위해 중심합성계획법(central composite design, CCD)을 기반으로 한 반응표면 분석법(Response Surface Methodology) 을 수행하였다. 세가지 독립변수로는 tryptone, yeast extract, 그리고 NaCl이 조사되었다. 반응값에 대하여 분산분석을 실시한 결과 결정계수(R2)는 0.96이었으며 전체 모델에 대한 유의확률이 0.0001로 매우 높은 유의성을 지님을 확인하였다. 반응표면분석법을 통하여 얻어진 B. subtilis NC1의 cellulase 활성을 위한 최적화 배지의 각 변수 농도는 tryptone 2.5%, yeast extract 0.5%, 그리고 NaCl 1.0%로 예측 되었다. 최적화 배지에서의 B. subtilis NC1의 cellulase 활성을 검증한 최적화를 실시하기 이전인 대조구의 cellulase 활성 0.5U/ml와 비교하면 24% 활성이 향상된 0.62U/ml의 높은 활성을 보였다.

고추역병 방제능이 있는 식물성장촉진 균주 Bacillus licheniformis K11의 cellulase 유전자의 cloning 및 효소 특성 조사 (Cloning of the Cellulase Gene and Characterization of the Enzyme from a Plant Growth Promoting Rhizobacterium, Bacillus licheniformis K11)

  • 우상민;김상달
    • Applied Biological Chemistry
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    • 제50권2호
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    • pp.95-100
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    • 2007
  • 식물성장 촉진호르몬인 auxin, 식물병원성 진균을 방제하는 siderophore 그리고 cellulase를 동시에 생산하는 PGPR균이자 생물방제균인 Bacillus licheniformis K11의 cellulase의 유전자를 PCR을 이용해 pUC18에 재조합 후 E. coli DH5${\alpha}$에 cloning하였으며, 이 형질전환 된 균주를 E. coli DH5${\alpha}$(pCW 77)라 하였다. 형질전환 균주 E. coli DH5${\alpha}$(pCW 77)는 B. licheniformis K11의 1.6kb 유전자를 포함하며, 이 cellulase는 1,479 bp, 499개의 amino acid가 암호화된 것으로 추정된다. 형질전환균주가 생산하는 cellulase(CelW)는 lac 프로모터를 이용해 발현되었으며, CMC-SDS-PAGE의 방법으로 약 55 kDa의 분자량을 확인하였다. B. licheniformis K11의 cellulase는 4종의 대표적인 Bacillus spp. 들이 생산하는 cellulase의 아미노산 배열이 97% 이상 일치하였다. CelW는 carboxymethyl-cellulose(CMC) 뿐만 아니라 불용성 섬유소인 Avicel, Filter paper(Whatman$^{\circledR}$ No. 1)는 물론이고 고추역병균 P. capsici의 건조 cell wall도 분해하였다. CMC를 기질로 60$^{\circ}C$에서 효소활성이 가장 높았으며, 최적 pH는 pH 6.0이었다. 그리고 CoCl$_2$ 또는 MnSO$_4$ 첨가시 활성이 2배 이상 증가하였지만, FeCl$_3$ 또는 HgCl$_2$ 첨가 시는 활성이 20% 이하로 떨어졌고, SDS와 sodium azide 등 여러 화학 저해제들을 첨가하여도 87% 이상의 활성을 유지하였다. 이 결과들은 B. licheniformis K11이 식물뿌리에 근권 microflora형성의 중요한 요인으로 작용할 수 있고, 생물방제력을 발휘하는 식물병원성 진균의 세포벽 분해 cellulase 기능 연구를 가능케 하여 식물병의 생물학적 방제 연구에 기초가 될 것이라 생각된다.

인삼 종자의 성숙과 후숙 과정에서 배유세포내 섬유소 가수분해효소의 분포 및 기능 (Localization and Function of Cellulase in Endosperm Cells of Panax ginseng Seeds during Maturation and After-ripening)

  • 유성철
    • Journal of Plant Biology
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    • 제36권4호
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    • pp.327-335
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    • 1993
  • The active sites, intracellular transport, function of cellulase in association with the disintegration of the storage materials of the endosperm cells during seed maturation and after-ripening of Panax ginseng C.A. Meyer seeds were studied by electron microscopy. Cytochemical activities of the cellulase occurred in protein bodies and vesicles of endosperm cells in seed with red seed coat. In after-ripening seed, the activities were strongly found in the cell wall of endosperm near the umbiliform layer and on neighbouring vesicles, so it is assumed that these cells begin to be decomposed. Cellulase activities were initiated before the decomposition of storage materials. But, no activity was observed in the umbiliform layer, so it is suggested that cellulase lose its activity after the completion of lysis process.

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텐셀소재의 fibrillation에 관한 연구(제I보) -셀룰라제로 처리한 텐셀의 물성변화를 중심으로- (A Study on Fibrillation of Tencel Material(Part I) -Based on Change of Properties of Tencel by Cellulase-)

  • 정영희;송경헌;양진숙
    • 한국의류학회지
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    • 제25권3호
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    • pp.507-515
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    • 2001
  • Tencel is a high quality cellulosic fiber, which is controlled and processed by an environmentally sound route. But, the point about tencel and the reason why it achieves its unique touch in its finished state is that it is a fibrillating fiber. That means it can take a great deal of punishment during the finishing process, which raise the fiber to produce the characteristics handle, without destroying the cloth. The aim of cellulase treatment is to improve appearence of tencel, plus the fiber ends protruding from the fabric surface. But enzymatic hydrolysis can weakens the fiber ends and changes the properties of fabrics. This study examined about the changes of properties according to several conditions and effect of cellulase to fibril of tencel. The results are as follows. The weight loss was occurred, tensile strength was decreased, softness was increased. And cellulase treatment reduced amount of fibril.

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셀룰라아제에 의한 텐셀직물의 가공에 있어서 호제의 영향 (Effect of Sizing Agent on the Enzymatic Finishing of Tencel Fabric by Cellulase)

  • 최창남;황태연;고봉국;박원규;변수진;이웅의;정상귀;조성용
    • 한국염색가공학회지
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    • 제15권1호
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    • pp.8-14
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    • 2003
  • The effect of pasting agent on the defibrillation of Tencel fabric was investigated. It was evaluated by the weigth loss of fabric when the fabric was treated with cellulase containing various kinds of pasting agents. The surface appearance of Tencel fabric was checked by SEM. Under the treatment condition without pasting agent, the weight loss of fabric was high at pH 5.0 and $60^\circ{C}$. This means that the cellulase activity was high at this condition. By increasing the concentration of carboxymethyl cellulose(CMC), the weight loss of fabric was decreased monotonously. This tendency was not appeared in other pasting agents. CMC is synthesized by the reaction of chloroacetic acid and cellulose. The glucose units may be remained after the reaction. So, it was considered that the degradation of glucose unit in Tencel was decreased, because cellulase had to attack both Tencel and CMC.

Preparation and Characteristics of Alkaline -active Cellulases from Coprinaceae

  • Lee, Jung-Kyung;Eom, Tae-Jin
    • 펄프종이기술
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    • 제31권3호
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    • pp.68-76
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    • 1999
  • Coprinus cinereus 2249 producing alkaline-active cellulase was screened from 29 species of Corpinaceae and constitutively produced alkaline carboxymethyl cellulase (CMCase) and filter paper cellulase (Fpase). When cultivated at pH 9.0, 25$^{\circ}C$ and 5 days, copnnus cinereus 2249 produced higher alkaline activity on 0.5% CMC, 2% wheat bran as carbon source and 0.5% peptone, 0.05% yeast extract as nitrongen source compared with other culture conditions. The level of cellulase production was higher in the presence of wheat bran than in the presence of CMC. The optimum temperature and pH for alkaline -active cellulase activity weas 50$^{\circ}C$ and 9, 0, respectively.

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Cytochemical and Immunocytochemical Study on the Cellulase Activity in the Accessory Glands of the Digestive System of the Oriental Land Snail, Nesiohelix

  • Jeong, Kye-Heon;Lee, Young-Seok;Shim, Yun-Bo
    • 한국패류학회지
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    • 제15권2호
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    • pp.81-92
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    • 1999
  • The histochemical, cytochemical, and immunocytochemical investigations were conducted to find out the cellulase activity in the accessory glands of the digestive system of the oriental land snail Nesiohelix samarangae under the LM, SEM, and TEM. The cellulase activity was shown in the epithelium of th digestive gland by labelling with the immunogold (protein-A gold) particles. The epithelial cells showing the cellulase activity were Type 1 and Type 3 cells out of five types of the epithelial cells of the digestive gland. None of epithelial cells of the mucus gland and the salivary gland and the salivary gland were not labeled with the immunogold particles.

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섬유소(纖維素)의 당화(糖化) (Enzymatic Hydrolysis of Cellulose)

  • 이계준
    • 생약학회지
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    • 제7권2호
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    • pp.85-93
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    • 1976
  • Since cellulose is the only organic material that is annually replenishable in very large quantities, we must explore ways to utilize it as a source of energy, food and chemicals. For the utilization of this resource, it is first enzymatic hydrolyzed to glucose, then the glucose can be used as a food, converted single cell protein by microorganism, fermented to clean burning fuel and other chemicals. Cellulolytic enzyme, cellulase, consists of two or three major components, $C_1-cellulase$, $C_x-cellulase$ and ${\beta}-glucosidase$. $C_x-cellulase$ are fairly common but $C_1-cellulase$ are quite rare. Trichoderma viride is the best source of active cellulose, especially $C_1-enzyme$. Saccharification rate of cellulose in greatly influenced by the degree of crystallinity and extent of lignification. But by the pretreatment the substrate with cellulose swelling agent, delignifying reagent and physical treatment, the degree of saccharification is enhanced. Thus, glucose syrups of 2 to 10% concentration are realized from milled newspaper. The enzymatic hydrolysis of such energy rich material, such as cellulose, to glucose is technically feasible and practically achievable on a very large scale.

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Cellulase Activity of Symbiotic Bacteria from Snails, Achatina fulica

  • Kim, Jon Young;Yoon, Sae Min;Kim, Yeong-Suk
    • Journal of the Korean Wood Science and Technology
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    • 제43권5호
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    • pp.628-640
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    • 2015
  • Cellulase is the key enzyme in the use of cellulose-based biomaterials. Because of its structure, cellulose is difficult to be degraded by enzymes. In order to utilize cellulose-based biomaterials efficiently, evolutionary wisdom of how to use enzymes accurately and harmoniously in a biological system is needed, such as the cellulose digestive system in animals. In this study, the symbiotic bacteria from snails, Achatina fulica, were identified and their cellulase activity was evaluated. The 16S rRNA sequence analysis of 100 aerobic bacteria showed that they belonged to 9 genus and almost half of the bacteria were Lactococcus spp. Among 100 identified strains, only two Aeromonas sp. strains showed cellulase activity. Aeromonas sp. KMBS020 had both endo-${\beta}$-glucanase and ${\beta}$-glucosidase activities but Aeromonas sp. KMBS018 had ${\beta}$-glucosidase activity only. None of the 100 bacterial colonies had any cellobiohydrolase activity.

Trichoderma reesei QM 9414를 이용한 섬유소 분해효소 생산조건에 관한 연구 (Studies on Cellulase Production by Trichoderma reesei (QM 9414))

  • 김종민;류두영
    • 미생물학회지
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    • 제16권4호
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    • pp.141-147
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    • 1978
  • In order to increase the productivity of cellulolytic enzymes, medium composition and culture conditions were studied. When cellulose powder (Avicel) supplemented with rice straw was used as carbon source, productivity of ${\beta}-glucosidase$ was increased by about 3 times compared with the runs with only cellulose powder as a carbon source. In this case no negative effects on the production of CMC enzyme activity and filter paper activity was found. For the production of celulolytic enzymes using T. reesei QM 9414, casitone was found to be a good nitrogen source compared with other sources studied, such as peptone, yeast extract, tryptone, and casein. The highest cellulase activity was attained when 0.3% glucose and 0.01% Tween 80 were supplemented to the standard medium of Rese. An adequate oxygen transfer rate was also found to be important to the cellulase fermentation and about 50 mmole of oxygen/liter/hour supported good cellulase biosynthesis during cellulase fermentation.

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