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Distribution and Favorable Conditions for Mycelial Growth of Cordyceps pruinosa in Korea (한국에 자생하는 붉은자루동충하초(Cordyceps pruinosa)의 분포와 균사생장에 적합한 조건)

  • Shin, Jae-Chul;Shrestha, Bhushan;Lee, Won-Ho;Park, Young-Jin;Kim, Soo-Young;Jeong, Gwang-Ryel;Kim, Ho-Kyung;Kim, Tae-Woong;Sung, Jae-Mo
    • The Korean Journal of Mycology
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    • v.32 no.2
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    • pp.79-88
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    • 2004
  • Cordyceps pruinosa grows upon dead pupae of Lepidoptera and produces one or $3{\sim}4$ club-shaped stromata per host. The stromata have distinct club-shaped head and long stalk. The length of stromata varies from $1{\sim}3\;cm$. Apical head consists of densely crowded semi-immersed perithecia, which are $360{\sim}400\;{\times}\;180{\sim}200\;{\mu}m$ in size. Asci are $150\;{\mu}m$ in length and $2.8{\sim}3\;{\mu}m$ in diameter. Ascospores, which are $124{\sim}141\;{\mu}m$ in length, have thin thread-like structures in the middle with part-spores attached on both sides. Each ascospore does not separate into part-spores after dispersal, but each part-spore germinates and together develops a colony. The imperfect form produces phialides of $15{\sim}24\;{\times}\;2{\sim}3\;{\mu}m$ size, with spherical or spindle shaped conidia of $4{\sim}6\;{\times}\;1.8{\sim}2.4\;{\mu}m$ size, The anamorph was identified as Mariannaea elegans Samson. YMA and SDAY agar media with pH 7 was produced abundant mycelial growth with high density. Best mycelial growth was observed when dextrin was used as a carbon source. Lactose, saccharose and sucrose also produced high mycelial growth. Peptone, yeast extract and tryptone produced abundant mycelial growth, when used as nitrogen sources. Highest mycelial growth and density was observed when C/N ratio was 1 : 1 at the concentration of 12.5 g/l each. $KH_2PO_4$ was the best mineral source for mycelial growth. Highest mycelial dry wt. was produced in YM and SDAY broths. Optimum inoculum for 100 ml of liquid broth was 6 mycelial discs. Similarly, optimum liquid culture period was 7 days.

Activities of the Hydrolytic Enzymes Produced by Plant Pathogenic Fungi, Sclerotium rolfsii, Sclerotinia Sclerotinia and Sclerotiorum, and Helminthosporium sigmoideum var. irregulare (수종의 식물병원균(흰비단병균$\cdot$균핵병균 및 좀검은 균핵병균)이 생산하는 가수분해효소의 활성)

  • Cho B. H.;Kim K.
    • Korean journal of applied entomology
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    • v.16 no.4 s.33
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    • pp.199-208
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    • 1977
  • Activities of various hydrolytic enzymes produced by three plant pathogenic fungi, Sclerotium rolfsii Sacc., Sclerotinia sclerotiorum (Lieb.) deBary and Helminthosporium sigmoideum var. irregulare Crallery et Tullius, were measured. Activties and amounts of the enzymes in mycelia, cultural filtrates, and sclerotia(except of sclerotia of H. sigmoideum var. irregulare) were estimated at various pH levels in order to find out optimal pH for their enzymatic activities. Enzymes such as cellulase (ex), invertase, xylanase, $\beta-amylase$, polymethylgalacturonase, polygalacturonase, phosphatase and protease were estimated. Culture solution for production of enzymes was prepared by adding of 10g, D-glucose, 1.3g $NH_4NO_3,\; 0.5g\; MgSO_4,\;7H_2O,\; and\; 1.0g\; KH_2PO_4$ into 1 liter of potato decoction plus 2ml of micro element solution consisting of 0.2mg. Fe, 0.2mg Zn, and 0.1mg Mn as the sulphates into 1 liter of distilled water. All tested mycelia and cultural filtrates were obtained from the cultures incubarted in previous solution for ten days at $25^{\circ}C$, and sclerotia were harvested from PDA plates of 3. days old, The crude enzyme solutions were prepared according to the method of Miyazaki etal. Ten days after incubation, activities of Cx produced by Scl. sclerotiorum were higher than those of the other fung and each of Cx from three fungi showed different pH optima, such as S. rolfsii and Scl. schlerotiorum in acid side (around pH 3.0), H. sigmoideum var. irregulare in neutral side (around pH 6.3). Invertase activities of S. rolfsii were 20 times higher than those of the other fungi in all samples. All tested fungi, however, showed no significant difference between the enzymatic activities of their cultural filtrate and mycelia and the activities in sclerotia of S. rolfsii and Scl. sclerotiorum were hardly recognized. There were multiple peaks on the xylanase activity curves of three fungi in terms of pH values. High activities of the xylanase were revealed in sclerotia of S. rolfsii and Scl. sclerotiorum, and in mycelia of H. sigmoideum var. irregulare. The highest activities of $\beta-amylase$ were shown both in mycelia and cultural filtrate of H. sigmoideum var. irregulae among the tested fungi, and their optimal pH was 6.2 in both mycelia and cultural filtrate. In the S. rofsii and Sel. sclerotiorum, however, the activities of cultural filtrates were higher than those of the other fungi, and optimal pH was 3.0 and 6.2 for cultural filtrate and both mycelia and sclerotia, respectively. Activities of PMG were high in cultural filtrates of all tested fungi, especially in Scl. sclerotiorum and H. sigmoideum var. irregulare. Mycelia of themalso showed the considerable activities. Optimal pH for enzymatic activities were variable with thekind of fungi or with the samples measured. The highest activities of PG were presented by mycelia of S. rolfsii and Scl. sclerotiorum. $9.l\mu /min.\; and\; 9.5\mu g/min.$, respectively. Optimal pH for activity of PG in mycelia was around 4.5 in S. rolfsii and around 3.0 in Scl. sclerotiorum. Phosphatase of S. rolfsii and Scl. sclerotiorum was more active in acid side (optimal PH3. 5) and that of H. sigmoideum var. irregulare showed one peak each in acid, neutral and alkaline side. But the highest peak was at pH 9.5. Protease of all tested fungi was more active at pH 10.0, especially that of the cultural filtrate of H. sigmoideum var. irregualre.

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Optimization of Growth Medium and Poly-$\beta$-hydroxybutyric Acid Production from Methanol in Methylobacterium organophilum (메탄올로부터 Methylobacterium organophilum에 의한 Poly-$\beta$-hydroxybutyric Acid의 생산과 배지성분의 최적화)

  • Choi, Joon-H;Kim, Jung H.;M. Daniel;J.M. Lebeault
    • Microbiology and Biotechnology Letters
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    • v.17 no.4
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    • pp.392-396
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    • 1989
  • Methylobacterium organophilum, a facultative methylotroph was cultivated on a methanol as a sole carbon and energy source. The cell growth was affected by the various components of minimal synthetic medium and the medium composition was optimized with 0.5% (v/v) methanol at pH 6.8 and at 3$0^{\circ}C$. The maximum specific growth rate of M. organophilum was achieved to 0.26 hr$^{-1}$ in the optimized medium which has following composition: Methanol, 0.5% (v/v):(NH$_4$)$_2$SO$_4$, 1.0g/l:KH$_2$PO$_4$, 2.13g/l:KH$_2$PO$_4$, 1.305g/ι:MgSO$_4$.7$H_2O$. 45g/l and trace elements (CaCl$_2$.2$H_2O$, 3.3mg:FeSO$_4$.7$H_2O$, 1.3mg:MnSO$_4$.4$H_2O$, 130$\mu\textrm{g}$:ZnSO$_4$.5$H_2O$, 40$\mu\textrm{g}$:Na$_2$MoO$_4$.2$H_2O$, 40$\mu\textrm{g}$:CoCl$_2$.6$H_2O$, 40$\mu\textrm{g}$:H$_3$BO$_3$, 30$\mu\textrm{g}$ per liter). By the limitation of nitrogen and deficiency of Mn$^{+2}$ or Fe$^{+2}$, the cell growth was significantly repressed. Methanol greatly repressed the cell growth and the complete inhibition was observed at concentration above 4% (v/v). In order to overcome the methanol inhibition and to prevent the methanol limitation, intermittent feeding of methanol was conducted by a D.O.-stat technique. PHB production by M. organophilum was stimulated by deficiency of nutrients such as NH$_{4}^{+}$, SO$_{4}^{-2}$, $Mg^{+2}$, $K^{+}$, or PO$_{4}^{-3}$ in the medium. The maximum PHB content was obtained as 58% of dry cell weight under deficiency of potassium ion in the optimized synthetic medium.

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An Comparison of Storability of Several Cultivars Chicon Grown in Different Regions (품종과 재배지역에 따른 치콘의 저장성 비교)

  • Kang, Ho-Min;Kim, Hyuk-Su;Seo, Hyun-Taek;Won, Jae-Hee
    • Journal of Bio-Environment Control
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    • v.18 no.3
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    • pp.291-296
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    • 2009
  • This study was conducted to compare the storability of 6 chicory cultivars for producing chicon; 'Vintor', 'Focus', 'Metafora', 'Kibora', 'Nobus', and 'Redoria Red' grown in 2 regions: Chuncheon (plain region) and Pyeongchang (high land region). Chicons were produced from chicory roots that grown for 120days and then stored for over 120days at $2^{\circ}C$ and 90% of RH conditions. To produce chicon, chicory root was forced at $18^{\circ}C$ for 22days with suppling the nutrient solution ($KNO_30.54g{\cdot}L^{-1}$, $Ca(NO_3)_2\;1.02g{\cdot}L^{-1}$, $MgSO_4\;0.36g{\cdot}L^{-1}$, $KH_2PO_4\;0.21g{\cdot}L^{-1}$, $K_2SO_4\;0.10g{\cdot}L^{-1}$, pH 7.0). Chicons produced from 6 different chicory cultivars packed with $25{\mu}m$ ceramic film and stored for 25days at $8^{\circ}C$. The fresh weight of chicon in MAP was maintained to 99.5% of pre-storage weight. The fresh weight of 'Redoria Red' was lowest in all cultivars, and that of Chuncheon region cultivated treatment was lower than Pyeongchang treatment. The $CO_2$ and $O_2$ concentration in chicon MAP were 2% and $10{\sim}17%$. There were not significantly different among cultivars and between regions, although 'Redoria Red' cultivar showed highest $CO_2$ and lowest $O_2$ concentrations. The ethylene concentration in chicon MAP was $1.0{\mu}{\iota}{\cdot}{\iota}^{-1}$ and also didn't show any significant difference among all treatments. Chicon detoriorated visual quality with appearing russet spotting that result from ethylene gas. The visual quality of 'Redoria Red' cultivar decreased faster than the other cultivars. 'Metafora', 'Focus', and 'Kibora' maintained higher firmness of their leaf than the others, and the firmness was higher grown in Pyeongchang region cultivated treatments than in Chuncheon region.

Studies on the Utilization of Alcohol Distillers' Waste. Part 1. Production of Saccharomyces cerevisiae Cells from Alcohol Distilled Waste of Wheat-flour. (주정폐액의 이용에 관한 연구(제1보) 소맥분 주정폐액을 이용한 Saccharomyces cerevisiae의 배양)

  • Yoo, Ju-Hyun;Oh, Doo-Hwan;Yang, Yung
    • Microbiology and Biotechnology Letters
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    • v.2 no.2
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    • pp.83-88
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    • 1974
  • The water pollution by waste water is one of the important issue and the short of animal feed is too, in Korea. So, this experiment is accomplished to treat alcohol distillers' waste by micro-organisms and planning to produce yeasts, which can be used as animal feed, pharmacy and condiments. 1. The raw material, alcohol distillers' waste, of this experiment consists of insoluble solids (residue) and filterate (supernatant). The residue contains 33.08% of crude protein, 19.96% of total sugar. and 2.06% of ash, respectively. On the other hand the flterate through the Toyo filter paper No. 5C, contains 2.48% of crude protein, 1.54% of reducing sugar, and 0.43% of ash, respectively. 2. Optimum pH of the basal medium for the growth of Saccharomyces cerevisiae YF-1 is 4.0. Optimum culture condition of this is as follows : when 0.43g of urea, 0.43g of potassium phosphate monobasic, and 0.21g of magnesium sulfate are added to the 100m1 of basal medium. Optimum temperature and optimum incubation time are 30$^{\circ}C$ and 24-28 hrs. 3. Under these conditions, the maximum yield of dry yeast is 1.38% to the medium. 4. The composition of dry yeast, produced under these conditions, is as follows: crude protein, 56.96%, lipid, 1.30%. total sugar, 6.53%, and ash 9.62%.

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Optimization of Culture Media for Solid-state Culture of Pleurotus ferulae

  • Cha Wol-Suk;Choi DuBok;Kang Si-Hyung
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.5
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    • pp.369-373
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    • 2004
  • In order to elucidate the possibility of artificial production of p. ferulae by solid-state culture, the optimization of culture conditions was carried out. When $NH_4H_2PO_4$ and $CaCO_3$ were used in the cultures using test tube with 30 g of Populus sawdust at $25^{\circ}C{\pm}1$ in the dark, the favored mycelial growth was observed with $1\%$ of $NH_4H_2PO_4$ and the production of polysaccharide was 7.85 mg/100 mg of mycelium with $1\%$ of $CaCO_3$. The mixtures of $80\%$ of Populus Sawdust and $20\%$ of rice bran at $60\%$ of water content were determined to be optimal for the production of fruiting bodies in the sawdust culture. When three treatments containing various ratios of garlic powder were conducted, yields of fruiting bodies were drasti[ally higher than those of Synthetic mixture without garlic powder The highest yield (143 g/bag) was obtained with $7\%$ garlic powder. The yield of synthetic mixture containing $7\%$ of garlic powder was $83\%$ higher than that of Sawdust culture. The reason why garlic powder did support growth was not clear but it is possible that garlic powder might contain effective components for the formation of fruiting body. The optimal synthetic mixture composition consisted of cotton seed $77\%$, lime $6.4\%,\;K_2HPO_4\;0.2\%,\;KH_2PO_4\;0.2\%,\;CaHPO_4\;0.2\%$, corn flour $4\%$, wheat flour $5\%$, and garlic pow-der $7\%$.

Control Effect of Phosphate Compounds on Phytophthora Blight and Anthracnose of Pepper (고추 역병과 탄저병에 대한 인산염 화합물의 방제 효과)

  • Park, Seojeong;Kim, Heung Tae
    • The Korean Journal of Pesticide Science
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    • v.21 no.1
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    • pp.26-32
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    • 2017
  • Antimicrobial activity of 6 phosphate compounds as $H_3PO_3$, $H_3PO_4$, $K_3PO_4$, $K_2HPO_4$, $KH_2PO_4$ and $NH_4H_2PO_4$ against Phytophthora capsici JHAW 1-2 and Colletotrichum acutatum JC24 was investigated in this study. Inhibitory effect on zoospore release, zoosporangia germination and zoospore germination was superior than mycelial growth. Among 6 compounds, $H_3PO_3$ and $H_3PO_4$ showed the best antimicrobial activity against P. capsici JHAW 1-2. Diseases controlling activity of the phosphate compounds tested on seedling and fruit of pepper against Phytophthora blight was also better than those against anthracnose. When $H_3PO_3$ was applied to the pepper seedlings at a concentration of $100{\mu}g\;mL^{-1}$, severe phytotoxicity was occurred. However, with applying $10{\mu}g\;mL^{-1}$ of $H_3PO_3$ showed 100% the disease control efficacy. In case of $100{\mu}g\;mL^{-1}$ $H_3PO_4$ application showed excellent antimicrobial activity against P. capsici JHAW 1-2, and 56.7% of the disease control efficacy with no phytotoxicity. To investigate the control efficacy against anthracnose, conidia suspension was inoculated with non-wound and wound inoculation method on pepper fruit. Among 6 compounds, only $100{\mu}g\;mL^{-1}$ of $H_3PO_3$ and $H_3PO_4$ had a activity of more than 70%, but the control activity on other treatments was minimal or unacceptable.

Establishment of Optimal Production Conditions of Transglucosidase Produced by Aspergillus niger (Aspergillus niger가 생산하는 transglucosidase의 최적 생산 조건 확립)

  • Lee, Jun-Yeob;Gang, Seongho;Kim, Jong-Sik;Chung, Chungwook
    • Journal of Life Science
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    • v.28 no.8
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    • pp.969-976
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    • 2018
  • In this study, transglucosidase (TG), an enzyme produced by Aspergillus niger, synthesized isomaltooligosaccharide from ${\alpha}-(1{\rightarrow}4)$ linked substrates. The highest TG-producing A. niger KCTC6913 was selected from six kinds of species, and optimized TG producing conditions were established. Five different carbon sources (potato starch, sweet potato starch, corn starch, wheat starch, and dextrin) and three different nitrogen sources (yeast extract, malt extract, and beef extract) were tested to establish the carbon and nitrogen sources favorable for TG production. Measurements of TG activity after an initial culture at pH 5.0 for 15 days revealed that potato starch and yeast extract, which are basic culture media, resulted in the highest TG activity. In addition, A. niger KCTC6913 increased TG production under aerobic conditions and a controlled carbon/nitrogen ratio. In conclusion, to evaluate TG activity in the established optimal medium, it is confirmed that the basal and potato dextrose broth medium were used as a control, and the highest TG production was measured, which was highlighted in the established optimal medium.

Evaluation for the simultaneous Removal of Organic Matters and Nutrients by the RBC and tapered Aeration Processes with Bacillus sp. for the high Strength of Dairy Wastewater (바실러스 미생물을 이용한 고농도 유가공 폐수처리에 있어서 유기물질과 영양염류의 동시제거에 대한 평가)

  • Lee, Sang-Ho
    • Journal of Korean Society of Water and Wastewater
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    • v.24 no.2
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    • pp.195-202
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    • 2010
  • The evaluation of organic matters and nutrients removal was investigated for the synthetic wastewater and the high strength of dairy wastewater. Two different systems were performed for this research. System A composing of a single RBC with tapered aeration was fed with the synthetic wastewater for 74 days with 173L/day of influent, 200% of internal return and 100% of sludge return for the period 1 and 2. The feed conditions were maintained 346L/day of influent, 50% of internal return and sludge return for the period 3. The dairy wastewater was introduced to evaluate treatment efficiency for system B composing of dual RBCs and tapered aeration tanks for 50 days of experimental run time, in which hydraulic rates were maintained at the constant ratios of 346L/day, 50% of internal return and 50% of sludge return. The spiral string media made of nylon fibre was attached by Bacillus sp. in RBC for both systems. The specific area of string media was $1.4m^2$/m and biomass was maintained at the concentrations of 23g/m. The synthetic wastewater was supplied by 1,800mg/L of glucose, 500mg/L of $NH_4Cl$, and 500mg/L of $KH_2PO_4$ to system A. The dairy wastewater was supplied to system B from dairy production plant. The average influent concentrations were 1,334mg/L of BOD, 2,014mg/L of CODcr, 160mg/L of T-N, and 12mg/L of T-P in system A. The average influent concentrations of parameters were 1,006 mg/L for BOD, 1,875mg/L for $COD_{cr}$, 51.6mg/L for T-N and 8.9mg/L for T-P in system B. Results indicated that removal efficiencies of BOD and $COD_{cr}$ were more than 90% however, the removal efficiency of T-N was 87%, and that of T-P was 82% for system A. Removal efficiencies were 98.5% of BOD, 91.3% of nitrogen and 89% of phosphorus for system B. The removal efficiencies of organic matters, T-N and T-P were higher in system B than in system A. The effluent quality issued by the stringent national legislations for the discharge of the high strength of dairy products wastewater can be improved using sequential RBCsand tapered aeration reactors rather than a single RBC and tapered aeration reactors with Bacillus sp.

Optimal Conditions for the Laccase Production from Fomitopsis pinicola Mycelia (Fomitopsis pinicola 균사체로부터 Laccase의 최적생산조건)

  • Park, Naomi;Park, Sang-Shin
    • Microbiology and Biotechnology Letters
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    • v.37 no.1
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    • pp.62-68
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    • 2009
  • The culture conditions to maximize the production of laccase (EC 1.10.3.2) from Fomitopsis pinicola mycelia were investigated. Among the tested media for the enzyme production, mushroom complete medium (MCM ; 2% dextrose, 0.2% peptone, 0.2% yeast extract, 0.05% $KH_2PO_4$, and 0.05% $MgSO_4{\cdot}7H_2O$) showed the highest activity of the enzyme. To optimize the culture condition for the laccase activity, influence of various carbon and nitrogen sources was investigated in MCM. Among various carbon and nitrogen sources, 2% glucose and 0.4% peptone showed the highest production of the enzyme, respectively. For the phosphorus and inorganic source, 0.05% $NaH_2PO_4$ and 0.05% $CaCl_2$ were best for the enzyme activity. The enzyme production was reached to highest level after the cultivation for 8 days at $25^{\circ}C$. Native polyacrylamide gel electrophoresis (PAGE) followed by the laccase activity staining using 2, 2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) as the substrate was performed to identify the laccase under culture conditions studied. Zymogram analysis of the culture supernatant showed a laccase band with molecular mass of 52 kDa. The optimum pH and temperature for the enzyme activity were $80^{\circ}C$ and pH 3.0.