• Title/Summary/Keyword: CMCase activity

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Ergosterol Contents and Enzymatic Characteristics of Lentinula edodes During Culture and Fruiting Periods (표고 균주의 배양 기간과 자실체 발생 기간에 따른 에르고스테롤 변화와 효소적 특성)

  • Kim Myungkil;Yoon Kabhee;Bak Wonchull;Park Hyun;Choi Joonweon;Lee Jaewon;Lee Bonghun
    • Journal of Korea Foresty Energy
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    • v.23 no.2
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    • pp.21-28
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    • 2004
  • Three different strains of Lentinula edodes, Sanlim 5-Ho, Sanlim 6-Ho and Nongki 3-Ho, were cultured in the sawdust media of Mongolian oak(Quercu mongolica Fisch) for 90 days under dark and light conditions(each 30 days) and fruiting period(30 days). Weight loss of sawdust media was determined after fungal cultures and the contents of ergosterol in fungal mycelia were quantified by HPLC analysis followed by solvent extraction. Compared with the two other fungal strains$(8\%)$, weight loss of Sanlim 5-Ho was slightly lowered to $7\%$. The level of ergosterol content, a parameter for fungal growth, was continuously enhanced in Sanlim 5-Ho for dark and light incubation periods. However, Sanlim 6-Ho and Nongki 3-Ho recorded the maximized fungal growth under light condition. In fruiting periods the ergosterol contents were lowered in the three strains. Intra- and extracellular enzymes during cultural and fruiting periods were also characterized. The activity of Mn-peroxidase and laccase, which are characteristics enzymes for white rot fungi as lignin degrading enzymes, were determined as a high level overall the periods. As cellulose degrading indicators, the activity of CMCase, avicelase, xylanase and glucanase were detectable in initial incubation period.

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Production of Cellulolytic Enzymes by Trichoderma harzianum FJ1 in Solid State Fermentation. (Trichoderma harzianum FJ1의 고체상태배양에 의한 섬유소분해효소의 생산)

  • 유승수;김경철;김성준
    • Microbiology and Biotechnology Letters
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    • v.31 no.3
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    • pp.257-263
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    • 2003
  • The cellulases production in solid state fermentation (SSF) of Trichoderma harzianum FJ1 with high cellulases productivity using cellulosic wastes was investigated. Physical and chemical conditions of the fermentation, such as moisture content, initial pH, and composition of mixed substrate (wine waste, rice straw, and soybean flour) on FPase (Filter paper activity) production were examined. The enzyme production was optimized in the conditions of moisture content of 70%, pH 5.0, 3$0^{\circ}C$, and 1:1:1 composition of mixed substrate containing wine waste, rice straw, and soybean flour. The highest activities of FPA, CMCase, Xylanase, $\beta$-glucosidase, and Avicelase in the optimized culture conditions were 15.2, 69.1, 83.9, 29.2, and 4.2 unit/g-SDW in 5 day cultivation, respectively. Economical and efficient production of cellulolytic enzymes by T harzianum FJ1 using cellulosic wastes in solid state fermentation will contribute to the biological saccharification of cellulosic wastes with enormous potential resource value in future.

Isolation and physiological characteristics of cellulolytic bacteria (섬유소 분해세균의 분리 및 생리적인 특성)

  • Kwon, Oh-Jin;Chung, Yung-Gun
    • Applied Biological Chemistry
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    • v.37 no.4
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    • pp.226-233
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    • 1994
  • Three hundred and one cellulolytic bacterial were isolated from the 148 screening sources such as decomposed wood, soil, compost and leaf mold. Among them, strain KL-6 was found to have the highest of cellulase activity, and identified as species belonged to the genus Cellulomonas. Strain KL-6 was decompose up to 90% of the filter paper (whatman No. 1) substrate within 50 hours, and showed the colony halo formation (11 cm). The activities of CMCase (67 unit/ml), FPase (70 unit/ml) and ${\beta}-glucosidase$ (0.68 unit/ml) were obtained when this strain was cultured for 50 hrs at $30^{\circ}C$. Glucose was not found in detectable amounts at the FP medium. The optimum composition of nutrient medium for the cell growth by strain KL-6 was sucrose 0.5%, yeast extract 0.1%, $(NH_4)_2HPO_4\;0.1%$, $K_2HPO_4\;0.1%$, $MgSO_4{\cdot}7H_2O\;0.01%$, $CaCl_2\; 0.01%$, NaCl 0.6%, $CaCO_3\;0.1%$ and the optimum pH and temperature were 7.0 and $30^{\circ}C$, respectively.

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Studies on the Isolation, Purification and Characterization of a Cx Enzyme Produced by Pyricularia oryzae, $C-7^{+t}$ (도열병균에서 추출한 Cx효소의 순화 및 특성에 관한 연구)

  • Kim, Sang-Ho;Kim, W.S.
    • The Korean Journal of Mycology
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    • v.10 no.2
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    • pp.67-73
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    • 1982
  • The $(NH_4)_2\;SO_4$ (70%) treated crude enzymes from the culture filtrates of the$C-7^{+t}$ strain of Pyricularia oryzae which was grown on 2% CMC (carboxymethyl cellulose) for 8 days at $28^{\circ}C$ were chromatographied on Sephadex G-150 and DEAE-Sephadex A-25 columns. From the chromatography, three fractions of CMCase$(C_x)$ was examined using Na-CMC as substrate. The $C_x$ enzyme activity was optimal at pH 6.0 and $40^{\circ}C$, stable up to $40^{\circ}C$. The values of Km and Vmax of the enzyme were $2.8{\times}10\;mM$ and 5.9m moles/hour, respectively. The molecular weight determined by Sephadex G-150 column chromatography was around 80,000. Approximately sevenfold purified $C_x$ enzyme gave a single protein band on the polyacrylamide gel electrophoresis.

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Isolation and Identification of Hydrolytic Enzyme-producing Bacteria from Spent Mushroom Substrate (버섯부산물유래 가수분해효소분비 박테리아의 분리 및 동정)

  • Kim, Young-Il;Jeong, Se-Hyung;Seok, Joon-Sang;Yang, Si-Yong;Huh, Jung-Won;Kwak, Wan-Sup
    • Journal of Animal Science and Technology
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    • v.50 no.5
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    • pp.713-720
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    • 2008
  • This study was conducted to isolate and identify xylanase- and cellulase-producing thermophilic bacteria from stacked spent mushroom substrates and to determine the optimal medium conditions for their growth. Bacteria with the highest xylanase and CMCase activities were strain 3 and 201-7. Both of them were identified as Bacillus spp. and named Bacillus subtilis KU3 and Bacillus subtilis KU201-7. The optimal medium condition of Bacillus subtilis KU3 was obtained when 3%(w/v) of yeast extract and 1%(w/v) of maltose were used as nitrogen and carbon sources, respectively. That of Bacillus subtilis KU201-7 was obtained when 0.5%(w/v) of yeast extract and 0.5%(w/v) of CMC were used as nitrogen and carbon sources, respectively.

Cloning of Thermophilic Alkalophilic Bacillas sp. F204 Cellulase Gene and Its Expression in Escherichia coli and Bacillus subtilis (고온 알칼리성 Bacillus sp. F204의 Cellulase 유전자의 Escherichia coli 및 Bacillus subtilis에의 Cloning 및 발현)

  • Chung, Young-Chul;Kim, Yang-Woo;Kang, Shin-Kwon;Rho, Jong-Su;Park, Jae-Hyeon;Sung, Nack-Kie
    • Korean Journal of Food Science and Technology
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    • v.23 no.1
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    • pp.31-36
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    • 1991
  • Cellulase genes from thermophilic alkalophilic Bacillus sp. F204 a potent cellulase complex-producing bacterium, were cloned in Escherichia coli with pUC 19. Plasmids pBC191 and pBC192, isolated from transformants forming yellow zone around colony on the LB agar plate containing 0.5% carboxymethyl cellulose and ampicillin, contained 4.6 Kb and 5.8 Kb HindIII fragments, respectively. The 4.6 Kb insert of pBC191 had single sites for BamHI EcoRI, KpnI and pvuII. DNA hybridization and immunodiffusion studies showed that pBC191-encoded cellulase gene was homologous with that of host strain. pKC231, constructed by inserting 4.6 Kb insert of pBC191 at the HindIII site of pKK223-3, E. coli expression vector, and pGC711, constructed by inserting 4.6 Kb insert of pBC191 at the HindIII site of pGR71, E. coli and B. subtilis shuttle vector, had 3.2 times and 2.8 times as much cellulase activity as pBC191, respectively. Substrate specificity analysis showed that cellulases cloned were CMCase.

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Formation of Intergeneric Hybrids Between Aspergillus niger and Penicillium verruculosum by Nuclear Transfer (핵전이에 의한 Aspergillus niger와 Penicillium verruculosum F-3의 속간 잡종형성)

  • Yang, Young-Ki;Park, Yeol;Kim, Sung-Joon;Cheong, Hyeon-Suk;Lim, Chae-Young;Rhee, Young-Ha
    • Korean Journal of Microbiology
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    • v.31 no.1
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    • pp.1-8
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    • 1993
  • lntergeneric hybrids formed between Aspergillus niger and Penicillium verruculosum were obtained by nuclear transfer technique. Nuclei isolated from wild type and auxotrophic mutants of donor strains were transferred into the protoplasts of different auxotrophic mutants as recipient strains. Several auxotrophic mutants were isolated from conidiospores of the two strains mutagenized with ultraviolet and N-methyl-N'-nitrosoguanidine. Optimal conditions for formation of intergeneric hybrids were investigated. Frequencies of intergeneric hybrid formation by nuclear transfer were $7{\times}10^{5}~1{\times}10^{5}$. From observations of genetic stability. DNA content. nuclear stain and conidial size. it was suggested that their karyotypes are aneuploid. In addition. the hybrids possess the 1.1~2.3-fold higher cellulase activities than those of parental strains. It was also revealed that some hybrids had different isozyme patterns compared to those of parental strains by CMCase and $\beta$-glucosidase activity assays.

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Effects of Types of TMR on Rumen Fermentation Characteristics and Nutrients Digestibility in Sheep (유형별 완전혼합사료 급여가 반추위내의 발효성상 및 영양소 소화율에 미치는 영향)

  • Li, D.Y.;Ko, J.Y.;Choi, N.J.;Lee, Sang S.;Song, J.Y.;Lee, S.Y.;Park, S.H.;Sung, H.G.;Ha, J.K.
    • Journal of Animal Science and Technology
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    • v.45 no.5
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    • pp.805-812
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    • 2003
  • This study was conducted to examine effects of feeding dry TMR(DTMR), wet TMR(WTMR) and fermented TMR(FTMR) on rumen fermentation, enzyme activity and digestibility in the total tract of sheep. Three rumen cannulated sheep were used in a 3${\times}$3 latin square design. The present results showed that pH, NH3-N, total and individual VFA, A/P ratio and enzymes (CMCase, Xylanase and Protease) activity in the rumen were higher in WTMR and FTMR compared with DTMR. In addition, dry matter, organic matter, crude protein, ether extract, NDF and ADF digestibility in the total tract were also higher in WTMR and FTMR compared with DTMR. Therefore, the present results showed that WTMR and FTMR are better than DTMR for rumen fermentation and nutrients digestibility.

Isolation and Identification of Cellulomonas fimi, Characteristics of its Cellulase and Conversion of the Sawdust into Ethanol (Cellulomonas fimi의 분리(分離) 및 동정(同定), cellulase 특성(特性)과 톱밥의 Ethanol 전환(轉換))

  • Lee, Chan-Yong;Lee, Ke-Ho
    • Applied Biological Chemistry
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    • v.28 no.2
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    • pp.98-105
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    • 1985
  • In the sheep and cattle's rumen, facultative anaerobic cellulolytic bacteria were isolated by using Hungate's roll tube technique. In the 21 isolated species, one was screened by its strong cellulolytic activity and identified as Cellulomonas fimi C-14 by investigate morphological, cultural, physiological characteristics and electron microgram. Optimum conditions of the cell growth and enzyme production were pH 6.5 an $30^{\circ}C$, Thiamine and biotin support a good growth of C. fimi C-14. In the enzyme activities, Crystalline cellulose hydrolyzing activity, CMCase activity and ${\beta}-glucosidase$ activity were 20.6, 226.6 and 0.56$(unit{\times}10^3/ml)$ at pH 6.0, $40^{\circ}C$. By addition of fungal cellulase, enzyme activity was increased. Simultaneous Saccharification Fermentation is better than two step fermentation in ethanol yield with Saccharomyces cerevisiae DY2.

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Cellulose Utilization and Protein Productivity of Some Cellulolytic Fungal Co-cultures

  • Eyini, M.;Babitha, S.;Lee, Min-Woong
    • Mycobiology
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    • v.30 no.3
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    • pp.166-169
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    • 2002
  • Protein productivity by the cellulolytic fungi, Trichoderma viride(MTCC 800), Chaetomium globosum and Aspergillus terreus was compared in co-culture and mixed culture fermentations of cashewnut bran. Co-cultures were more effective in substrate saccharification, which ranged between $85{\sim}88%$ compared to the $62{\sim}67%$ saccharification shown by the monocultures. Maximum saccharification was induced by T. viride and C. globosum co-culture resulting in the highest 34% release of reducing sugars. The maximum 16.4% biomass protein and the highest protein productivity(0.58%) were shown by T. viride and A. terreus co-culture. A. terreus performed better in co-culture in the presence of T. viride rather than with C. globosum. Among the cellulolytic enzymes, FPase(Filter Paper Cellulase) activity was significantly higher in all the co-cultures and in the mixed culture than in their respective monocultures. Mixed culture fermentation involving all the three fungi was not effective in increasing the per cent saccharification or the biomass protein content over the co-cultures.