• Title/Summary/Keyword: Cellulases

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Metagenomic Analysis of Novel Lignocellulose-Degrading Enzymes from Higher Termite Guts Inhabiting Microbes

  • Nimchua, Thidarat;Thongaram, Taksawan;Uengwetwanit, Tanaporn;Pongpattanakitshote, Somchai;Eurwilaichitr, Lily
    • Journal of Microbiology and Biotechnology
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    • v.22 no.4
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    • pp.462-469
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    • 2012
  • A metagenomic fosmid library was constructed from genomic DNA isolated from the microbial community residing in hindguts of a wood-feeding higher termite (Microcerotermes sp.) collected in Thailand. The library was screened for clones expressing lignocellulolytic activities. Fourteen independent active clones (2 cellulases and 12 xylanases) were obtained by functional screening at pH 10.0. Analysis of shotgun-cloning and pyrosequencing data revealed six ORFs, which shared less than 59% identity and 73% similarity of their amino acid sequences with known cellulases and xylanases. Conserved domain analysis of these ORFs revealed a cellulase belonging to the glycoside hydrolase family 5, whereas the other five xylanases showed significant identity to diverse families including families 8, 10, and 11. Interestingly, one fosmid clone was isolated carrying three contiguous xylanase genes that may comprise a xylanosome operon. The enzymes with the highest activities at alkaline pH from the initial activity screening were characterized biochemically. These enzymes showed a broad range of enzyme activities from pH 5.0 to 10.0, with pH optimal of 8.0 retaining more than 70% of their respective activities at pH 9.0. The optimal temperatures of these enzymes ranged from $50^{\circ}C$ to $55^{\circ}C$. This study provides evidence for the diversity and function of lignocellulose-degrading enzymes in the termite gut microbial community, which could be of potential use for industrial processes such as pulp biobleaching and denim biostoning.

Studies on Cellulases of Penicillium chrysogenum (Penicillium chrysogenum에서 추출한 cellulase에 관한 연구)

  • 노명희
    • Korean Journal of Microbiology
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    • v.17 no.1
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    • pp.42-48
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    • 1979
  • Cellulases were isolated from both healthyl(PC) and virus0infected penicillum chrysogenum(PCV) in the wheat bean culture, and some properties of the enzymes were studied. 1) At $37^{\circ}C$, pH 6, 4 and 6day's culture the maximal enzyme yield was obtained in both PC and PCV. 2) The optimum temperature for the PC cellulase was at $50^{\circ}C$, and that for the PCV enzyme was the same. 3) The optimum pH for the PC enzyme was at 5.0, whereas the PCV enzyme was at 6.0, indicating that they are isozymes. 4) When Na-CMC was used as a substrate, PC enzyme was twice as high as the activity of PCV enzyme.

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Characterization of Cellulases from Schizophyllum commune for Hydrolysis of Cellulosic Biomass (Schizophyllum commune에 의한 Cellulase 생산 및 섬유소계 바이오매스의 당화를 위한 효소적 특성)

  • Kim, Hyun-Jung;Kim, Yoon-Hee;Cho, Moon-Jung;Shin, Keum;Lee, Dong-Heub;Kim, Tae-Jong;Kim, Yeong-Suk
    • Journal of the Korean Wood Science and Technology
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    • v.38 no.6
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    • pp.547-560
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    • 2010
  • The optimum culture condition of Schizophyllum commune for the cellulase production and its enzymatic characteristics for saccharification of cellulosic biomass were analyzed. S. commune secrets ${\beta}$-1,4-xylosidase (BXL) and cellulases, including endo-${\beta}$-1,4-glucanase (EG), cellobiohydrolase (CBH), and ${\beta}$-glucosidase (BGL). The optimum reaction temperature for all cellulases was $50^{\circ}C$ and the thermostable range was $30{\sim}40^{\circ}C$C. The optimum reaction pH for all cellulases was 5.5 in a range of temperature from $0^{\circ}C$ to $55^{\circ}C$. The best nutritions for the cellulase production of S. commune among tested nutrients were 2% cellulose for the carbon source and corn steep liquor or peptone/yeast extract for the nitrogen source without vitamins. The environmental culture condition for the cellulase production was 5.5~6.0 for pH at $25{\sim}30^{\circ}C$. The enzyme activities of EG, BGL, CBH, and BXL were 3670.5, 631.9, 398.5, and 15.2 U/$m{\ell}$, respectively, after concentration forty times from the culture broth of S. commune which was grown at the optimized culture condition. Alternative filter paper unit assay showed 11 FPU/$m{\ell}$ enzyme activity. The saccharification tests using cellulase of S. commune showed the low saccharification rate on tested hardwoods but a high value of 50.5% on cellulose, respectively. The saccharification rate (50.5%) of cellulose by cellulase produced in this work is higher than 45.7% in the commercial enzyme (Celluclast 1.5L, 30 FPU/g, glucan).

Empirical Evaluation of Cellulase on Enzymatic Hydrolysis of Waste Office Paper

  • Park, Enoch Y.;Ikeda, Yuko;Okuda, Naoyuki
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.5
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    • pp.268-274
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    • 2002
  • Enzymatic hydrolysis of waste office paper was evaluated using three commercial cellulases, Acremonium cellulase, Meicelase, and Cellulosin T2. Varying the enzyme loading from 1 to 10% (w/w) conversion of waste office paper to reducing sugar was investigated. The conversion increased with the increase in the enzyme loading: in the case of enzyme loading of 10% (w/w), Acremonium cellulase yielded 79%conversion of waste office paper, which was 17% higher compared to Meicelase, 13% higher than that of Cellulosin T2. Empirical model for the conversion (%) of waste office paper to re-ducing sugar (x) was derived from experimental results as follow, x = $kE^{m}t^{(aE+b)}$ where k, m, a, and b de-note empirical constants. E indicates initial enzyme concentration.

The beating effect of high crystalized nonwood fibers treated with low-molecular weighted waste celulase in the papermaking processes (Cellulase생산공정중 발생되는 저분자량 분포도의 폐효소류 처리가 고결정화된 배목재 섬유소의 고해에 미치는 영향)

  • 김병현;신종순;강영립;박병권;이성구
    • Journal of the Korean Graphic Arts Communication Society
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    • v.18 no.1
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    • pp.121-139
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    • 2000
  • This study is to test the possibility of applying the low-molecular weighted waste cellulase, which is produced in the process of cellulase production, to paper making. After experimenting on high-crystallized non-wood fibers with beating catalyst. I got the result that the condition for the optimal effect is temperature 40~6$0^{\circ}C$, the time 90min to 120min, pH 5.0 to 6.0, the enzyme contents 0.3% and that the effect of beating such as slight reduction of fiver viscosity, increase of water retention value(WRV) and shortening of fiber length was increased with waste cellulase. Through this process, the density, folding endurance, tensile strength and burst strength of paper was remarkably increased, which is inferred to result from the increased flexibility of fiber by individual characteristics of non-wood fiber, which was high-crystallized by penetrated low-molecular weight cellulases in the fiber.

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Intein-mediated expression of Trichoderma reesei Cellobiohydrolase I Cellulose Binding Domain in E. coli (Intein을 이용한 대장균에서의 Trichoderma reesei 유래의 Cellobiohydrolase I 섬유소 결합 도메인의 발현)

  • Choi, Shin-Geon
    • Journal of Industrial Technology
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    • v.36
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    • pp.33-37
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    • 2016
  • Cellulose binding domains (CBDs) of cellulases are thought to assist in the hydrolysis of insoluble crystalline cellulose. To gain sufficient amount of CBDs, the self-cleavable intein tag was used for expression and purification of Trichoderma reesei cellobiohydrolase I CBD in E. coli. Synthetic CBD genes, CBD or linker-CBD were cloned into expression vector pTYB11. Recombinant CBDs were successfully purified by intein mediated purification with an affinity chitin-binding domain. The final yields of recombinant CBD and linker-CBD were 3.2 mg/L and 1.4 mg/L, respectively. The functional bindings of recombinant CBDs were confirmed by Avicel binding experiments. The simple and easy purification method using self-cleavable intein tag can be further used in pretreatment of crystalline cellulose or characterization of engineered CBDs.

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The Role of Rumen Fungi in Fibre Digestion - Review -

  • Ho, Y.W.;Abdullah, N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.1
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    • pp.104-112
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    • 1999
  • Since the anaerobic rumen fungi were discovered in the rumen of a sheep over two decades ago, they have been reported in a wide range of herbivores fud on high fibre diets. The extensive colonisation and degradation of fibrous plant tissues by the fungi suggest that they have a role in fibre digestion. All rumen fungi studied so far are fibrolytic. They produce a range of hydrolytic enzymes, which include the cellulases, hemicellulases, pectinases and phenolic acid esterases, to enable them to invade and degrade the lignocellulosic plant tissues. Although rumen fungi may not seem to be essential to general rumen function since they may be absent in animals fed on low fibre diets, they, nevertheless, could contribute to the digestion of high-fibre poor-quality forages.

Extremozymes: A Potential Source for Industrial Applications

  • Dumorne, Kelly;Cordova, David Camacho;Astorga-Elo, Marcia;Renganathan, Prabhaharan
    • Journal of Microbiology and Biotechnology
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    • v.27 no.4
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    • pp.649-659
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    • 2017
  • Extremophilic microorganisms have established a diversity of molecular strategies in order to survive in extreme conditions. Biocatalysts isolated by these organisms are termed extremozymes, and possess extraordinary properties of salt allowance, thermostability, and cold adaptivity. Extremozymes are very resistant to extreme conditions owing to their great solidity, and they pose new opportunities for biocatalysis and biotransformations, as well as for the development of the economy and new line of research, through their application. Thermophilic proteins, piezophilic proteins, acidophilic proteins, and halophilic proteins have been studied during the last few years. Amylases, proteases, lipases, pullulanases, cellulases, chitinases, xylanases, pectinases, isomerases, esterases, and dehydrogenases have great potential application for biotechnology, such as in agricultural, chemical, biomedical, and biotechnological processes. The study of extremozymes and their main applications have emerged during recent years.

Impact of electron beam irradiation on enzymatic saccharification of yellow poplar (Liriodendron tulipifera L) (전자빔 조사 처리가 백합나무 효소 당화에 미치는 영향)

  • Shin, Soo-Jeong;Sung, Yong-Joo;Han, Gyu-Seong;Cho, Nam-Seok
    • 한국신재생에너지학회:학술대회논문집
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    • 2008.05a
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    • pp.198-201
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    • 2008
  • The electron beam irradiation was applied as a pretreatment of the enzymatic hydrolysis of yellow poplar with doses of 0$\sim$450 kGy. The higher irradiation dose resulted in the more degradation of hardwood biomass not only from carbohydrates but also from lignin. This changes originated from the irradiation resulted in the better response to enzymatic hydrolysis with commercial cellulases (Celluclast 1.5L and Novozym 342). The more improvement on enzymatic hydrolysis by the irradiation was found in the xylan than in the cellulose of yellow poplar.

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Enzymatic saccharification of rice straw, a lignocellulosic biomass by the extracellular enzyme from KMU001 (KMU001 조효소에 의한 목질계 바이오매스, 볏짚의 효소당화성에 관한 연구)

  • Kim, Yeong-Suk;Lee, Young-Min;Cha, Chang-Jun;Yoon, Jeoung-Jun
    • 한국신재생에너지학회:학술대회논문집
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    • 2008.05a
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    • pp.225-228
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    • 2008
  • This study shows that lignocellulosic biomass saccharification work has been carried out with rice-straw by the extracellular enzyme from KMU001, and the enzymes produced in 5%(w/v) wood biomass were characterized by protein and various enzyme activity measurements. Several cellulases such as Endoglucanase(EG), $\beta$-D-1,4-Glucosidase(BGL), Cellobiohydrolase(CBH), and $\beta$-D-1,4-Xylanase (BXL) were detected. Saccharification of rice-straw by the enzyme yielded about 233mg/g of glucose after 48hrs.

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