• Title/Summary/Keyword: cellulase activities

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Development of a Microbial Consortium with High Cellulolytic Enzyme Production (섬유소 분해 효소의 고생산을 위한 복합균주 개발)

  • 오영아;김경철;유승수;김성준
    • KSBB Journal
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    • v.17 no.4
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    • pp.381-387
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    • 2002
  • A filamentous fungus, strain FB01 showing high $\beta$-glucosidase activity was isolated from a compost. This fungus was cocultured with Trichoderma viride to enhance the productivity of $\beta$-glucosidase by changing inoculation time of the fungus. The microbial consortium showed higher cellulolytic enzyme production than T. viride alone. The maximal enzyme production was obtained when the microbial consortium was cultured at 30$\^{C}$ and pH 6.0 for 10 days with the activities of CMCase, $\beta$-glucosidase, and avicelase of 2.0, 0.8, and 0.2 U/mL, respectively. These enzyme activities were 2, 4, and 2 times as high as those of CMCase, p-glucosidase, avicelase from T. viride, respectively, indicating that a synergistic interaction appeared between T. viride and strain FBOI . The serial subcultures with pH control increased $\beta$-glucosidase production about 3.2 times. Enzyme production using ricestraw as a carbon source showed that the activities of CMCase, $\beta$-glucosidase, and avicelase were 3.69, 0.76, 0.17 U/mL, respectively, and $\beta$-glucosidase activity was 1.5 times higher than that of T viride.

Cultural Characteristics of Ectomycorrhizal Mushrooms

  • Jeon, Sung-Min;Ka, Kang-Hyeon
    • 한국균학회소식:학술대회논문집
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    • 2015.11a
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    • pp.16-16
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    • 2015
  • Ectomycorrhizal (ECM) mushrooms play a major role in plant growth promotion through symbiotic association with roots of forest trees. They also provide an economically important food resource to us and therefore they have been studied for their artificial cultivation for decades in Korea. We have secured bio-resources of ECM mushrooms from Korean forests and performed their physiological studies. To investigate the cultural characteristics, the fungi were cultured under different conditions (medium, temperature, pH of the medium, inorganic nitrogen source). More than 90% of total 160 strains grew on three solid media (potato dextrose agar, PDA; sabouraud dextrose agar, SDA; modified Melin-Norkrans medium, MMN). The rate of mycelial growth on malt extract agar (MEA) was lower than those of three media (PDA, SDA, MMN). None of the Tricholomataceae strains grew on MEA. Many strains of ECM mushrooms were able to grow at the temperature range of $15{\sim}25^{\circ}C$ on PDA, while they showed poor growth at $10^{\circ}C$ or $30^{\circ}C$. In particular, the growth rates of both Gomphaceae and Tricholomataceae were significantly lower at $10^{\circ}C$ than at $30^{\circ}C$. The optimal pH of many strains was pH 5.0 when they cultured in potato dextrose broth (PDB). Fifty-seven percent of tested strains grew well on medium containing ammonium source than nitrate source. Many strains of Tricholomataceae showed a notable growth on ammonium medium than nitrate medium. Twenty-three percent of strains preferred nitrate source than ammonium source for their mycelial growth. The production and activity of two enzymes (cellulase and laccase) by ECM fungi were also assayed on the enzyme screening media containing CMC or ABTS. Each strains exhibited different levels of enzymatic activities as well as enzyme production. The number of laccase-producing strains was less than that of cellulase-producing strains. We found that 77% of tested strains produced both cellulase and laccase, whereas 2% of strains did not produce any enzymes. The morphological characteristics of mycelial colony were also examined on four different solid media. Yellow was a dominant color in mycelial colony and followed by white and brown on all culture media. ECM mushrooms formed mycelial colonies with a single or multiple colors within a culture medium depending on the strains and culture media. The most common shape of mycelial colony was a circular form on all media tested. Other families except for Amanitaceae formed an irregular colony on MMN than PDA. All strains of Tricholomataceae did not form a filamentous colony on all media. The pigmentation of culture media by mycelial colonies was observed in more than 50% of strains tested on both PDA and SDA. The degree of pigmentation on PDA or SDA was higher than MMN and brown color was dominant than yellow color. The production of exudates from mycelial colony was higher on PDA than MMN. Brown exudates were mainly produced by many strains on PDA or SDA, whereas transparent exudates were mainly produced by strains on MMN. We observed the mycelial colonies with a single or multiple textures in just one culture plate. Wrinkled or uneven colony surfaces were remarkably observed in many strains on PDA or SDA, while an even colony surface was observed in many strains on MMN. Sixty percent of Tricholomaceae strains formed wrinkled surface on PDA. However, they did not form any wrinkle on MMN plate. Cottony texture was observed in mycelia colonies of many strains. Velvety texture was often observed in the mycelial colonies on SDA than PDA and accounted for 60% of Suillaceae strains on SDA.

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Changes in isoflavone content and quality characteristics of Cheonggukjang prepared with Bacillus subtilis HJ18-3 and KACC 15935 (Bacillus subtilis HJ18-3과 KACC 15935를 이용하여 제조한 청국장의 품질특성과 isoflavone 함량의 변화)

  • Lee, Kyung Ha;Choi, Hye Sun;Choi, Yoon Hee;Park, Shin Young;Song, Jin
    • Food Science and Preservation
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    • v.21 no.1
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    • pp.121-128
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    • 2014
  • This study was conducted in order to investigate the change of isoflavone composition (glycoside and bio-active aglycone), and to evaluate the quality characteristics of Cheonggukjang, which was prepared by different bacillus strains. After the 48-hour fermentation, the contents of daidzein, genistein, and glycitein in the Bacillus subtilis HJ18-3 have significantly increased up to approximately $89.06{\pm}3.59$, $10.36{\pm}0.28$, and $101.37{\pm}3.67ug/g$, respectively. The contents of daidzein, genistein, and glycitein in the Bacillus subtilis KACC 15935 were $38.88{\pm}5.39$, $12.58{\pm}2.14$, and $80.13{\pm}0.71ug/g$, respectively. The original content of daidzein was 3.96 ug/g, while genistein and glycitein were not measured. However, the contents of daidzen and genistein in HJ18-3 and in KACC 15935 were decreased. The ${\alpha}$-Amylase and cellulase activities of Chungkookjang in HJ18-3 were higher than in the KACC 15935. The contents of Chungkookjang in HJ18-3 were $29.70{\pm}11.66$ and $4861.3{\pm}388.07unit/g$, respectively. The amino type nitrogen contents and ammonia type nitrogen contents of Chungkookjang in KACC 15935 were higher than in the HJ18-3. These results suggested that it could be used to increase the bioactivity via fermentation with the Bacillus subtilis possessing a ${\beta}$-glucosidase activity with a view towards the development of functional foods.

Extraction of Polyphenols from Apple Peel Using Cellulase and Pectinase and Estimation of Antioxidant Activity (Cellulase와 Pectinase를 이용한 사과껍질 폴리페놀 추출 및 항산화 활성 평가)

  • Park, Min-Kyung;Kim, Cherl-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.5
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    • pp.535-540
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    • 2009
  • The effect of cellulolytic (Viscozyme) and pectolytic (Pectinex) enzyme treatments on extraction of total polyphenol and antioxidant activity of extract from apple peel have been examined. Extraction was carried out with a dosage of 0.1, 0.25, 0.5, 1 and 2% (v/v) of Viscozyme, Pectinex and Viscozyme+Pectinex at $30{\sim}50^{\circ}C$ for $12{\sim}24$ hours. Total polyphenol contents (mg/mL) of extracts obtained with 2% of Viscozyme, Pectinex or Viscozyme+Pectinex treatment for 12 hours were $0.30{\pm}0.02$, $0.16{\pm}0.01$, and $0.33{\pm}0.02$ at $30^{\circ}C$, $0.34{\pm}0.01$, $0.19{\pm}0.01$, and $0.35{\pm}0.02$ at $40^{\circ}C$ and $0.34{\pm}0.01$, $0.22{\pm}0.01$, and $0.38{\pm}0.02$ at $50^{\circ}C$ respectively. The result shows that Viscozyme was more effective than Pectinex at all experimental temperatures, and Viscozyme+Pectinex resulted in the highest phenolic content at $50^{\circ}C$. Antioxidant activities determined by DPPH, ABTS and FRAP assays were increased with concentrations of extracts produced by 2% of Viscozyme+Pectinex treatment, which ranged from 0.10 to 0.40 vit. C eq mM for $5{\sim}25\;mg$ of dried matters, from 0.09 to 0.28 vit. C eq mM for $1{\sim}5\;mg$ of dried matters, and from 0.06 to 1.85 $FeSO_4$ eq mM for $1{\sim}5\;mg$ of dried matters, respectively.

Biological Pretreatment of Softwood Pinus densiflora by Three White Rot Fungi

  • Lee, Jae-Won;Gwak, Ki-Seob;Park, Jun-Yeong;Park, Mi-Jin;Choi, Don-Ha;Kwon, Mi;Choi, In-Gyu
    • Journal of Microbiology
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    • v.45 no.6
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    • pp.485-491
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    • 2007
  • The effects of biological pretreatment on the Japanese red pine Pinus densiflora, was evaluated after exposure to three white rot fungi Ceriporia lacerata, Stereum hirsutum, and Polyporus brumalis. Change in chemical composition, structural modification, and their susceptibility to enzymatic saccharification in the degraded wood were analyzed. Of the three white rot fungi tested, S. hirsutum selectively degraded the lignin of this sortwood rather than the holocellulose component. After eight weeks of pretreatment with S. hirsutum, total weight loss was 10.7%, while lignin loss was the highest at 14.52% among the tested samples. However, holocellulose loss was lower at 7.81 % compared to those of C. lacerata and P. brumalis. Extracelluar enzymes from S. hirsutum showed higher activity of ligninase and lower activity of cellulase than those from other white rot fungi. Thus, total weight loss and changes in chemical composition of the Japanese red pine was well correlated with the enzyme activities related with lignin- and cellulose degradation in these fungi. Based on the data obtained from analysis of physical characterization of degraded wood by X-ray Diffractometry (XRD) and pore size distribution, S. hirsutum was considered as an effective potential fungus for biological pretreatment. In particular, the increase of available pore size of over 120 nm in pretreated wood powder with S. hirsutum made enzymes accessible for further enzymatic saccharification. When Japanese red pine chips treated with S. hirsutum were enzymatically saccharified using commercial enzymes (Cellulclast 1.5 L and Novozyme 188), sugar yield was greatly increased (21.01 %) compared to non-pre treated control samples, indicating that white rot fungus S. hirsutum provides an effective process in increasing sugar yield from woody biomass.

Basic study on the biological and physicochemical properties of burnt forest soil for the ecological restoration by organic waste (유기성폐자원을 이용한 산불토양의 생태학적 복원을 위한 토양의 생물학적, 물리화학적 기초특성연구)

  • Jung, Young-Ryul;Song, In-Geun;Kim, Young-Jun
    • Journal of the Korea Organic Resources Recycling Association
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    • v.13 no.1
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    • pp.79-89
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    • 2005
  • Forest soils were analyzed on their biological and physicochemical properties for the ecological restoration of burnt forest soil using organic wastes and proper microorganisms. Three kinds of soil samples were collected from undamaged soil(US), naturally restoring soil(NS) and artificially restoring soil(AS). All soil samples were sandy soil and acidic soil, ranged pH 5.34~5.78. Moisture content was higher in the soil of NS region. And the others were similar. Total organic matter and soluble sugar were higher at the surface, generally. Heterotrophic soil microbes were abundant at the surface soil of NS and subsoil of AS. Dehydrogenase, cellulase and phosphatase activities were higher at the NS soil. Especially, Dehydrogenase activity as primary index of soil microbial process showed high correlationship with moisture content(r=0.90, P < 0.05).

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The Effect of Saturated Fatty Acids on Cellulose Digestion by the Rumen Anaerobic Fungus, Neocallimatix frontalis C5-1

  • Ha, J.K.;Lee, S.S.;Gao, Z.;Kim, C.-H.;Kim, S.W.;Ko, Jong Y.;Cheng, K.-J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.7
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    • pp.941-946
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    • 2001
  • The effects of various concentrations of saturated fatty acids (SFA; caprylic, capric and stearic acids) on the growth of the anaerobic fungus, Neocallimastix frontalis C5-1 isolated from the rumen of a Korean native goat were investigated. At higher concentrations of fatty acids (0.1%, w/v), the addition of SFA strongly decreased filter paper (FP) cellulose digestion and polysaccharide-degrading enzyme activity. The sensitivity of the rumen anaerobic fungus to the added fatty acids increased in the following order: caprylic ($C_{8:0}$)>capric($C_{10:0}$)>stearic($C_{18:0}$) acid, although stearic acid had no significant (p<0.05) inhibitory effects at any of the concentrations tested. However, the addition of SFA at lower concentrations (0.01 and 0.001% levels), did not inhibit FP cellulose degradation and enzyme activity. Furthermore, although these parameters were slightly stimulated by the addition of SFA, they were not statistically different from control values. This is the first report examining the effects of fatty acids on anaerobic gut fungi. We found that the lower levels of fatty acids used in this experiment were able to stimulate the growth and specific enzyme activities of rumen anaerobic fungi, whereas the higher levels of fatty acids were inhibitory with respect to fungal cellulolysis.

고온. 호기법에 의한 중화요리잔반의 처리 과정에서의 중.고온균의 분리 및 특성

  • Yang, Jae-Kyung;Seo, Yong-Ki;Choi, Kyung-Min;Park, Eung-Roh;Whang, Key;Lee, Sung-Taik
    • Microbiology and Biotechnology Letters
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    • v.25 no.6
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    • pp.623-629
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    • 1997
  • The studies of isolation and physiological characteristics of mesophilic or thermophilic bacteria from thermophilic oxic process (TOP) treating Chinese restaurant wastes were conducted. Chinese restaurant wastes were consist of moisture; 75.8%, solids; 24.2% and ash; 0.49%. The volatile solid was about 99% of total dry solids. In wastes used in this experiment, there was content of crude protein; 4.47%, crude lipid; 3.56%, free sugar, 0.4% , crude starch; 10.34% and crude fiber 3.14%, respectively. And then it has about 4,970 kcal/kg-dry solid of Chinese restaurant wastes. From TOP treating the chinese restaurant wastes, 37 strains of mesophilic or thermophilic bacteria were primarily isolated using medium used for the isolation and among them 6 strains of thermophilic and 7 strains of mesophilic bacteria were selected by testing the activities of amylase, cellulase, protease and lipase. TB-1, TB-9 as thermophilic bacteria and MB-15-1, 15-2, MB23 as mesophilic bacteria having strong enzyme activity were selected among isolated strains. All selected strains reduced nitrate to nitrite and they utilized glucose, manose, manitol, and maltose as carbon source. From these MB15-2 was identified as Bacillus cereus, TB1, Bacillus licheniformis and TB9; Bacillus schlegelii.

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Characterization of a Phenazine and Hexanoyl Homoserine Lactone Producing Pseudomonas aurantiaca Strain PB-St2, Isolated from Sugarcane Stem

  • Mehnaz, Samina;Baig, Deeba Noreen;Jamil, Farrukh;Weselowski, Brian;Lazarovits, George
    • Journal of Microbiology and Biotechnology
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    • v.19 no.12
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    • pp.1688-1694
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    • 2009
  • A novel strain of fluorescent pseudomonad (PB-St2) was isolated from surface-sterilized stems of sugarcane grown in Pakistan. The bacterium was identified as Pseudomonas aurantiaca on the basis of 16S rRNA gene sequence analysis and results from physiological and biochemical characteristics carried out with API50 CH and QTS 24 bacterial identification kits. Assays using substrate-specific media for enzymes revealed lipase and protease activities but cellulase, chitinase, or pectinase were not detected. The bacterium was unable to solubilize phosphate or produce indole acetic acid. However, it did produce HCN, siderophores, and homoserine lactones. In dual culture assays on agar, the bacterium showed antifungal activity against an important pathogen of sugarcane in Pakistan, namely Colletotrichum falcatum, as well as for pathogenic isolates of Fusarium oxysporium and F. lateritium but not against F. solani. The antifungal metabolites were identified using thin-layer chromatography, UV spectra, and MALDI-TOFF spectra and shown to be phenazine-1-carboxylic acid (PCA), 2-hydroxyphenazine (2-OH-PHZ), and N-hexanoyl homoserine lactone (HHL) (assessed using only TLC data). The capacity of this bacterium to produce HCN and 2-OH-PHZ, as well as to inhibit the growth of C. falcatum, has not been previously reported.

Limits of Exogenous Fibrolytic Enzymes to Improve Digestion and Intake of a Tropical Grass

  • Assoumaya, C.;Boval, M.;Weisbecker, J.L.;Saminadin, G.;Archimede, H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.6
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    • pp.914-919
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    • 2007
  • The effect of the addition of exogenous fibrolytic enzymes (mainly xylanase and cellulase activities, 15 ml/15 kg of fresh forage), on intake, total tract digestibility and nylon bag degradability of a chopped fresh Digitaria decumbens grass was studied at 2 stages of regrowth ( 21 and 56-day old grasses). Moreover, comparisons between ground and chopped grass were done using the nylon bag degradability method. DM intake (g/kg $BW^{0.75}$) and organic matter total tract digestibility for control and enzyme treatments respectively were 69.1 vs. 65.9 (p>0.05) and 0.723 vs. 0.727 (p>0.05) with the 21-day old regrowth. Based on the same parameters, values for the 56-day old grass were 58.1 vs. 52.7 (p>0.05) and 0.621 vs. 0.591 (p>0.05). Nylon bag degradation at 24 h of the dry matter for control versus enzyme treatments were 0.653 vs. 0.70 (p<0.05) and 0.644 vs. 0.733 (p<0.0001) for the 21-day old chopped and ground forage respectively, whereas with the 56-day old grass, corresponding values were 0.321 vs. 0.392 (p<0.0001) and 0.463 vs. 0.481 (p>0.05). The positive impact of exogenous fibrolytic enzymes (EFE) on degradability of the young and ground pangola grass may suggest that in some cases, enzyme accessibility to potentially digestible cell wall is a limiting factor in their digestion.