• Title/Summary/Keyword: cellulase C

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Liquid Culture Enhances Protoplast Formation from the Auxotroph (Ser-) of lentinula edodes

  • Kim, Chae-Kyun;Kim, Byong-Kak
    • Archives of Pharmacal Research
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    • v.20 no.3
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    • pp.206-211
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    • 1997
  • The optimal conditions for the production and regeneration of the protoplasts from Lentinula edodes were studied. Protoplast formation from the mycelia of L. edodes which were cultured in liquid medium showed a significantly high yield compared with that of the mycelia which were cultured on cellophane covered agar media. A mixture of Novozyme 234 (15 mg/ml) and Cellulase Onozuka R10 (10 mg/ml) in 0.6 M mannitol (pH 4) was optimal lytic enzyme for the protoplast release. The optimal incubation time and mycelia age were 3.5-4 hours at $30^{\circ}C$ and 6-8 days, respectively. Regeneration frequency was 0.18% plated onto a medium containing 0.6 M sucrose, and 0.08% plated onto a medium containing mannitol. But hardly any regeneration was observed in the media containing NaCl, KCl, or $MgSO_{4}$ More than 90% of the protoplasts contained nuclei and the nucleus number per protoplast was 1.1. The DNA content per nucleus was 5.1 pg. The diameter of the protoplast was $3-5{\mu}m$ and it had a well defined cell structure.

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The Enzymatic Hydrolysis of Exploded Woody Biomass(I) -Effects of Lignin Contents and Cellulose Crystallinity on the Enzymatic Hydrolysis- (폭쇄처리(爆碎處理)된 목질계(木質系) Biomass의 산소가수분해(酸素加水分解)(I) -리그닌의 함량(含量)과 섬유소(纖維素)의 결정화도(結晶化度)가 산소가수분해(酸素加水分解)에 미치는 영향)

  • Park, Young-Ki;Oh, Jung-Soo
    • Journal of the Korean Wood Science and Technology
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    • v.21 no.3
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    • pp.53-60
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    • 1993
  • Substrates used were hardwood-Suwon poplar-(Populus alba${\times}$glandulosa L.) and softwood-pitch pine-(Pinus rigida M.). And these substrates were steam exploded then treated with sodium chlorite at 75$^{\circ}C$ with occasional stirring in order to obtain samples which had different lignin contents and crystallinity. And then this resulting samples incubated with a commercial cellulase derived from Trichoderma ressei. The contents of Klason lignin were decreased as the increasing of the ratio of sodium chlorite in the two species. The effect of hardwood was more effective than that of softwood in the same ratio of sodium chlorite. The minimum contents of Klason lignin were 0.8% and 5.1% respectively. And the crystallinities of cellulose were increased very little as increasing of the ratio of sodium clorite. The hydrolysis extent of the two species were increased as the increasing of delignification. Especially, the hydrolysis extent of hardwood was more higher than that of softwood. The maximum hydrolysis extent were 89.8% and 71.1%, respectively.

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Isolation and characterization of cellulolytic bacteria, Bacillus sp. EFL1, EFL2, and EFP3 from the mixed forest (혼효림으로부터 셀룰로오스분해 박테리아 분리 및 효소학적 특성규명)

  • Park, Hwa Rang;Oh, Ki-Cheol;Kim, Bong-Gyu
    • Journal of Applied Biological Chemistry
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    • v.61 no.1
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    • pp.59-67
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    • 2018
  • This study was conducted to isolate the cellulolytic bacteria able to grow on LB- Carboxymethyl cellulose (CMC) agar trypan blue medium from the mixed forest and Larix leptolepis stands. Three bacterial strains with high activity against both CMC and xylan were isolated. Both API kit test and 16S rRNA gene sequence analysis revealed that the three different isolates belong to the gene Bacillus. Therefore, the isolates named as Bacillus sp. EFL1, Bacillus sp. EFL2, and Bacillus sp. EFP3. The optimum growth temperature of Bacillus sp. EFL1, EFL2, and EFP3 were $37^{\circ}C$. The optimum temperature for CMCase and xylanase from Bacillus sp. EFL1 were $50^{\circ}C$. The optimum pH of Bacillus sp. EFL1 xylanase was pH 5.0 but the optimum pH of CMCase from Bacillus sp. EFL1 was pH 6.0. The optimum temperature of CMCase and xylanase from Bacillus sp. EFL2 was $60^{\circ}C$, respectively. The optimum pH of CMCase of Bacillus sp. EFL2 was 5.0, whereas xylanase showed high activity at pH 3.0-9.0. The optimum temperature for CMCase and xylanase of Bacillus sp. EFP3 was $50^{\circ}C$. The optimum pH for CMCase and xylanse was 5.0 and 4.0, respectively. CMCases from Bacillus sp. EFL1, EFL2, and EFP3 were thermally unstable. Although xylanase from Bacillus sp. EFL1 and EFP3 showed to be thermally unstable, xylanase from Bacillus sp. EFL2 showed to be thermally stable. Therefore, Bacillus sp. EFL2 has great potential for animal feed, biofuels, and food industry applications.

Effect of Feeding Chemically Treated Mustard (Brassica campestris) Straw on Rumen Fibre Degrading Enzymes in Sheep

  • Vaithiyanathan, S.;Raghuvanshi, S.K.S.;Mishra, A.S.;Tripathi, M.K.;Misra, A.K.;Prasad, R.;Jakhmola, R.C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.11
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    • pp.1610-1613
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    • 2003
  • The aim of the experiment was to study the changes in the activities of various rumen fibre degrading enzymes due to the feeding of chemically treated mustard (Brassica campestris) straw in sheep. Mustard straw (MS) (<5 cm particle size) was treated either with urea (4% (w/w), or with 2% sodium hydroxide (NaOH), or with alkaline hydrogen peroxide (2% NaOH and 1.5% hydrogen peroxide ($H_2O_2$)) and/or supplemented with 2% (w/w) urea. Seven maintenance type rations were prepared using MS (70 parts) with molasses (5 parts) and concentrate (25 parts). They were untreated MS (CMS), urea treated MS (UMS), urea supplemented MS (MSUS), alkali treated MS (AMS), alkali treated and urea supplemented MS (AMS-US), alkali $H_2O_2$ treated MS (AHMS) and alkali $H_2O_2$ treated and urea supplemented MS (AHMS-US). They were then compressed into a complete feed block with the help of block making machine. Forty two male hoggets of Malpura breed sheep were equally distributed into each treatment group and (were) offered feed and water ad libitum. At the end of 21 days of feeding trial, rumen liquor was collected through stomach tube from three animals in each group at 0 h, 4 h, 8 h, 12 h of post feeding. Results showed that the level of enzyme varied from 8.52 to 11.12, 40.85 to 50.37, 3.22 to 3.78, 2.09 to 2.77 and 31.44 to 44.24 units/100 ml SRL respectively for carboxymethyl cellulase (CMCase), $\alpha$-amylase, microcrystalline cellulase (MCCase), filter paper (FP) degrading enzyme and $\alpha$-glucosidase. Processing of MS affected the enzyme activities, in a way, that NaOH and AHP treatment significantly reduced CMCase and FP degrading enzyme. The effect of urea treatment showed an increase in the activity of MCCase and $\alpha$-glucosidase. But the supplementation of urea increased the activity of CMCase, FP degrading enzyme and $\alpha$-glucosidase. The CMCase, $\alpha$-amylase, $\alpha$-glucosidase activities were highest at 4hr whereas MCCase and FP degrading enzyme had maximum activities at 12 h post feeding Results suggested that MS might need longer time in the rumen for its effective degradation.

Studies on the promoted Aging of flue-cured leaf tobacco by cellulolytic Enzyme and Nicotinophiles (섬유소 분해효소 및 니코틴 분해세균을 이용한 잎담배의 발효촉진효과)

  • 이태호;성낙계
    • Journal of the Korean Society of Tobacco Science
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    • v.13 no.2
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    • pp.5-20
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    • 1991
  • For the quality enhancement of harvested-year leaf tobacco to the quality of 2-year naturally aged leaf tobacco, cellulose and nicotine degradative bacteria were isolated and identified. Effects of artificial fermentation treated cellulase and nicotine degradative bacteria on the quality of leaf tobacco were investigated from the chemical and sensory points of view. 1, Changes in chemical composition of leaf tobacco resulted from the addition of cellulase extracted from Cellulomonas sp. [3ml(${\mu}{\textrm}{m}$ D-glucose/ml. mil-1) of enzymes solution 11009 of leaf tobacco] and nicotine degradative bacteria, Pseudomonas sp. 2ml(IX109 cells$\div$ 100g of leaf tobacco), and subsequently fermented at 40${\mu}{\textrm}{m}$$^{\circ}C$, 65% R. H. for 40 days are as follows : 1) Content of crude fiber decreased 12% It took 9 min, 53 sec. to reach full combustion in control group but took only 7 min. 47 sec. in the treated group, taking almost equal time to 2-year naturally aged leaf tobacco(7 min. 35sec.). 2) Light intensity of control group was 60.96% with bright lemon color but that of treated leaf tobacco accounted for 47.69 with orange to dark brown color series, which was almost equal to the value, 45.69, of 2-year naturally aged leaf tobacco. 3) Linoleic acid, serving mild taste among organic acids, amounted to 1.llmg/g in control group but increased to 1.35m9/9 in the treated leaf tobacco, identical to the content(1.35mg/g) of 2-year naturally aged leaf tobacco. 4) Content of solanone, on of the typical leaf tobacco flavor compounds, accounted for 2.95% in control group but increased to 2.87% in treated group. 5) Methyl furan, useful flavor compound in smoke composition, accounted for 17.6$\mu\textrm{g}$/cig. in control group but increased to 25.9$\mu\textrm{g}$/cig. in treated group. However, acroleine decreased from 69.3$\mu\textrm{g}$/cig. in control group to 58.6$\mu\textrm{g}$/cig. in treated group 2. In sonsory test, mild taste evaluation of control group scored 5.47 and treated group 7.93 which was evaluted almost equal to the value(8.00) of 2-year naturally aged leaf tobacco. Aroma evaluation of control group scored 5.60, treated group 8.20, and 2-year naturally aged leaf tobacco 8.33. In addition, total harmony taste of control group showed 5.67, treated group 8.07 (p<0.01), and 2-year naturally aged leaf tobacco 8.00. From these results, it can be said that quality of treated leaf tobacco is not inferior to that 2-year naturally aged leaf tobacco.

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Hydrolysis of Agricultural Residues and Kraft Pulps by Xylanolytic Enzymes from Alkaliphilic Bacillus sp. Strain BK

  • Kaewintajuk Kusuma;Chon Gil-Hyong;Lee Jin-Sang;Kongkiattikajorn Jirasak;Ratanakhanokchai Khanok;Kyu Khin Lay;Lee John-Hwa;Roh Min-Suk;Choi Yun-Young;Park Hyun;Lee Yun-Sik
    • Journal of Microbiology and Biotechnology
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    • v.16 no.8
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    • pp.1255-1261
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    • 2006
  • An alkaliphilic bacterium, Bacillus sp. strain BK, was found to produce extracellular cellulase-free xylanolytic enzymes with xylan-binding activity. Since the pellet-bound xylanase is eluted with 2% TEA from the pellet of the culture, they contain a xylan-binding region that is stronger than the xylan-binding xylanase of the extracellular enzyme. The xylanases had a different molecular weight and xylan-binding ability. The enzyme activity of xylanase in the extracellular fraction was 6 times higher than in the pellet-bound enzyme. Among the enzymes, xylanase had the highest enzyme activity. When Bacillus sp. strain BK was grown in pH 10.5 alkaline medium containing xylan as the sole carbon source, the bacterium produced xylanase, arabinofuranosidase, acetyl esterase, and $\beta$-xylosidase with specific activities of 1.23, 0.11, 0.06, and 0.04 unit per mg of protein, respectively. However, there was no cellulase activity detected in the crude enzyme preparation. The hydrolysis of agricultural residues and kraft pulps by the xylanolytic enzymes was examined at 50$^{\circ}C$ and pH 7.0. The rate of xylan hydrolysis in com hull was higher than those of sugarcane bagasse, rice straw, com cop, rice husk, and rice bran. In contrast, the rate of xylan hydrolysis in sugarcane pulp was 2.01 and 3.52 times higher than those of eucalyptus and pine pulp, respectively. In conclusion, this enzyme can be used to hydrolyze xylan in agricultural residues and kraft pulps to breach and regenerate paper from recycled environmental resources.

Elimination and Utilization of Pollutants - Part I Microbiological Clarification of Industrial Waste and Its Utilization as Feed Resources - (환경오염원(環境汚染源)의 제거(除去)와 그 이용성(利用性)에 관(關)한 연구(硏究) - 제(報I)1보(第). 미생물(微生物)에 의(依)한 산업폐수(産業廢水)의 정화(淨化) 및 사료자원개발(飼料資源開發)에 개(開)하여 -)

  • Lee, Ke-Ho;Lee, Kang-Heup;Park, Sung-O
    • Applied Biological Chemistry
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    • v.23 no.1
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    • pp.64-72
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    • 1980
  • Industrial wastes from pulp and food plants were treated with microorganisms to clarify organic waste-water and to produce cells as animal feed, and results were summarized as follows. (1) Waste-water from pulp, beer, bread yeast, and ethanol distillation plants contained $1.4{\sim}1.5%$ of total sugar, $0.25{\sim}0.35%$ nitrogen, and biological oxygen demand (BOD) was $400{\sim}25,000$, chemical oxygen demand (COD), $500{\sim}28,000$, and pH, $3.8{\sim}7.0$. The BOD and COD were highest in waste-water from ethanol distillation plants among others. (2) Bacterial and yeast counts were $4{\times}10^4-1{\times}10^9,\;2{\times}10^2-7{\times}10^4/ml$ in waste-water. (3) Bacteria grew better in pulp waste and yeasts in beer, bread yeast, and ethanol distillation waste. (4) Saccharomyces cerevisiae SAFM 1008 and Candida curvata SAFM 70 were the most suitable microorganisms for clarification of ethanol distillation waste. (5) When liquid and solid waste from ethanol distillation were treated with microbial cellulase, xylanase, and pectinase, solid waste was reduced by 36%, soluble waste was increased, and recuding sugar content was increased by 1.3 times which provided better medium than untreated waste for cultivation of yeasts. (6) Optimum growth conditions of the two species of yeast in ethanol distillation waste were pH 5.0, $30^{\circ}C$, and addition of 0.2% of urea, 0.1% of $KH_2PO_4$ and 0.02% of $MgSO_4$. (7) Minimum number of yeast for proper propagation was $1.8{\times}10^5/ml$. (8) C. curvata70 was better than cerevisae for the production of yeast cells from ethanol distillation waste treated with microbial enzymes. (9) S. cerevisiae produced 16 g of dried cell per 1,000ml of ethanol distillation waste and reduced BOD by 46%. C. curvata produced 17.6g of dried cell and reduced BOD by 52% at the same condition. (10) Yeast cells produced from the ethanol distillation waste contained 46-52% protein indicating suitability as a protein source for animal feed.

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Ecological Studies on the Transition of Sheath Blight of Rice in Korea (한국(韓國)에서의 벼 잎집무늬마름병 발생변동(發生變動)에 관(關)한 생태학적(生態學的) 연구(硏究))

  • Yu, Seung-hun
    • Korean Journal of Agricultural Science
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    • v.4 no.2
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    • pp.283-316
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    • 1977
  • In an attempt to obtain a basic information to develop an effective integrated system of controlling sheath blight of rice in Korea, the transition of this disease, the variation of cultural characters and pathogenicity of the pathogen, environmental conditions affecting the disease outbreak and varietal resistance have been investigated. 1. Rice sheath blight which has been minor disease in the past was widely spread, especially since 1971. This disease has promptly spread all over the country and infected 65.2% of total rice growing area in 1976. Various factors are considered to be related to such transition of this disease. Above all, increace of application of nitrogenous fertilizer, early season and earlier cultivation of rice, introduction of more susceptible "Tongil" varieties etc. must be important factors influencing the outbreak of this disease. 2. Great variations in cultural characteristics-such as mycelial growth rate, color of the medium, amount of the aerial mycelium, shape and color of the sclerotia- and in the pathogenicity of isolates of the pathogen, Thanatephorus cucumeris Dank were observed. The optimum temperature for mycelial growth also varied with isolates, from $25^{\circ}C$ to $30^{\circ}C$. There were not necessarily any correlation between curtural characteristics and pathogenicity of isolates of Thanatephorus cucumens. 3. Mycelial grow th of isolates of Thanatephorus cucumens on the PDA medium were correlated with the air temperatures of the region where the isolates were collected. The isolates from the regions with high temperature grew well on PDA medium at $35^{\circ}C$ than those from the region with low temperature, on the other hand, the isolates from the regions with the low temperature grew well on the same medium at $12^{\circ}C$ than those from the regions with high temperature. 4. Pectin polygalacturonase (PG) and cellulase (Cx) were most active on the 3rd day after inoculation on the leaves of rice plant with Thanatephorus cucumeris, whereas pectin methylestrase (PE) was most active on the 4th day after inoculation. Relationship between the activities of PE of isolates and the strength of pathogenicity of isolates was obtained, but PG and cellulase activities were not correlated with pathogenicity of isolates. 5. The tolerence of sclerotia from in-vitro culture to low temperature varied with their water content, the dried cultural sclerotia were more tolerent than wet ones, Dried cultural sclerotia maintained almost 100% germinability for 45 days at $-20^{\circ}C$, whereas wet sclerotia lost viability at $-5^{\circ}C$. The germination ratio of the sclerotia after overwintering changed from 18% to 70% according to the water content of the test paddy fields and the ratio was low in wet paddy condition. 6. To investigate the host range of this fungi in and near paddy field, 17 weeds were inoculated with fungi. The lesions of sheath blight disease was obserbed on Sagittaria trifolia L., Echinochloa crusgalli P. Beauv., Monochoria vaginal is Presl, Polygonum Hydropiper L., Eclipta prostrata L., Digitaria sanguinalis Scapoli. 7. When the level of nitrogen applied was doubled over standard level, total nitrogen content in rice sheath increased, ami when silicate was applied, starch content in rice sheath decreased, inducing the rice plants more susceptible to sheath blight disease. Increased dressing of potash ferilizer reduced the incidence of sheat blight disease. 8. The percentage of infected stems in the early period increased more in the narrow hill plot than in the wide hill plot, but in the late period this tendency was inversed; the percentage of infected stems as well as severity in the wide hill plot increased more compared to the narrow hill plot, and the disease severity in the one plant per hill plot was also low. The number of stems in the wide hill plot was more than the number of stems in the narrow hill plot. This indicates that the microclimate, such as the relative humidity, in the narrow hill plot was more favorable for the development of this disease. 9. There was a high negative correlation between the disease severity of varieties to the sheath blight and the maturity of the varieties, that is, the early varieties were more susceptible than the late ones, and much-tillering varieties usually showed more infection than less tillering varieties. 10. No relationship was obtained between the percentage of infected stems in the early period and the severity after heading, whereas a distinct relationship was obtained between former and latter after Aug. 10.

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Effect of Additives, Storage Temperature and Regional Difference of Ensiling on the Fermentation Quality of Napier Grass (Pennisetum purpureum Schum.) Silage

  • Tamada, J.;Yokota, H.;Ohshima, M.;Tamaki, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.1
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    • pp.28-35
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    • 1999
  • The effects of addition of celulases (A cremonium cellulolyticus and Trichoderma viride, CE), a commercial inoculum containing lactic acid bacteria (Lactobacillus casei, LAB), fermented green juice (macerated napier grass with water was incubated anaerobically with 2% glucose for 1 day, FGJ) and glucose (G), and regional difference of ensiling on napier grass (Pennisetum purpureum Schum.) silage were studied by using 900 ml laboratory glass bottle silos under 30 and $40^{\circ}C$ storage conditions in 1995 and 1996. Experiment 1 was carried out to compare the addition of CE, LAB, FGJ and the combinations. Silages were stored for 45 days after ensiling. Experiment 2 studied the effects of applications of CE, LAB, FGJ and G. Experiment 3 was carried out using the similar additives as experiment 2 except for LAB. Silages were stored for 60 days in the experiments 2 and 3. Experiments 1 and 2 were done in Nagoya, and experiment 3 in Okinawa. Sugar addition through CE or G improved the fermentation quality in all the experiments, which resulted in a greater decrease in the pH value and an increased level of lactic acid, while butyric acid contents increased under $30^{\circ}C$ storage condition in CE addition. LAB and FGJ additions hardly affected the silage fermentation quality without additional fermentable carbohydrate. But the combination of LAB, FGJ and glucidic addition (CE and G) improved the fermentation quality. The effect of the regional difference of ensiling between temperate (Nagoya; $35^{\circ}$ N) and subtropical (Okinawa; $26.5^{\circ}$ N) zones on silage fermentation quality was not shown in the present study.

Characteristics of Mycelial Growth and Enzyme Activities of Mattirolomyces terfezioides Collected from Robinia pseudoacacia Forest in Korea (국내 아까시나무 숲에서 수집한 감자덩이버섯(Mattirolomyces terfezioides)의 균사 생장 특성과 효소 활성)

  • Jeon, Sung-Min;Wang, Eun-Jin;Ka, Kang-Hyeon
    • The Korean Journal of Mycology
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    • v.43 no.3
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    • pp.165-173
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    • 2015
  • Mattirolomyces terfezioides is a type of sweet truffles that predominantly form ectomycorrhizae with Robinia pseudoacacia. It is also worthy of artificial cultivation. This is the first report on characteristics of mycelial growth and enzyme activities of M. terfezioides collected from R. pseudoacacia forest in Korea. M. terfezioides showed the highest mycelial growth when cultured on potato dextrose agar (PDA) at $30^{\circ}C$ or in modified Melin-Norkran's liquid medium (pH 8.0). The biomass of M. terfezioides was higher in liquid medium containing nitrate-nitrogen than ammonium-nitrogen by 1.8 fold. The mycelia of M. terfezioides showed both carboxymethylcellulase and laccase activities on solid media for enzyme screening.