• Title/Summary/Keyword: Cultural mycelia

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A Study on the Purification and Characteristics of Branched-Chain Amino Acid Aminotransferase in Cultural Mycelia of Cordyceps militaris (번데기동충하초 균사 중의 Branched-Chain Amino Acid Aminotransferase의 분리정제 및 그 특성에 관한 연구)

  • Kim, Sung-Tae;Park, Chung-Oh
    • Korean Journal of Clinical Laboratory Science
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    • v.37 no.2
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    • pp.78-83
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    • 2005
  • The optimum conditions of Cordyceps militaris mycelial growth, purification and characteristics of branched-chain amino acid aminotransferase [BCAT(EC 2.6.1.42)] in this mycelium were studied. Optimum pH, temperature and medium of culture of mycelia were 5.5, $22.5^{\circ}C$ and Hamada medium (HM), respectively. BCAT in homogenate of this mycelia was precipitated by 20-40% saturated solution of ammonium sulfate and then purified by DEAE (diethylaminoethyl)-Sephadex A-50 column chromatography with linear concentration gradient and Sephadex G-200 gel filtration. A single band of purified enzyme was detected on SDS-PAGE (sodium dodecylsulfate-polyacrylamide gel electrophoresis). Optimum pH and temperature of BCAT were found to be 7.8 and $29^{\circ}C$, respectively. It showed activity toward L-leucine, L-isoleucine and L-valine as a substrate. The Km values of this enzyme for L-leucine were determined to be 5.88 mM for L-leucine.

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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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A STUDY ON THE MYCELIA CUL TURE AND GENETIC CHARACTERISTICS IN Lepista nuda (민자주방망이버섯(Lepista nuda)의 균사배양 및 유전적 특성에 관한 연구)

  • 김종봉;황성구
    • Journal of Life Science
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    • v.11 no.5
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    • pp.496-501
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    • 2001
  • This study was carried out to investigate the cultural characteristics and the genetic characteristics in Lepists nuda. The mycelia cultural characteristics which include the specificity for pH, carbon source, nitrogen source and medium were studied using petridish culture and liquid media culture respectively. The genetic characteristics were also investigated using random amplified polymorphic DNA (RAPD) analysis. To find out the mycelial growth rate by various medium, mycelia was cultured for 25 days in PDA, YM, MEA, GPB and Yamanaka. The Yamanaka was superior to the other media in supporting the mycelial growth. pH6 produced the best result in the test of an optimal pH. In experiment for optimal carbon source, starch showa 46.8$\pm$1.7mm of diameter of mycelial colony as the best and for nitrogen source, yeast extract showed a good effect as well. In investigation for genetic characteristics, Lepists nuda was amplified by 7 primers among 10 primers and Pleurotus ostreatus was also amplified by 7 primers. From the RAPD analysis between species, the band patterns were different.

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Reutilization of Enokitake Cultural Waste as Lentinus edodes Cultivation Substrate

  • Chai, Jung-Ki;Lee, Sung-Jin;Kim, Young-Ju;Wi, Kye-Moon
    • Plant Resources
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    • v.3 no.3
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    • pp.226-232
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    • 2000
  • The availability of enokitake cultural waste for Lentinus edodes cultivation was investigated, although hardwood sawdust has traditionally been used as a substrate for this fungus. Firstly, physiochemical characteristics of cultural waste were analysed. Secondly, mycelial growth characteristics and fruiting yields of L. edodes on waste treated in some methods were determined. Physiochemical characteristics of enokitake cultural waste showed that the millwaste complex was a little degraded by enokitake fungus and suggested the probability that most component lost by enokitake could be rice bran. Mycelia of L. edodes grew and fruited well on waste supplemented by fresh rice bran and Quercus sawdust although didn't on waste only. Mycelial growths of these fungi on waste were accelerated when supplemented by rice bran to the percent of 40(w/w) but decreased or suppressed at above ratios(30, 40%, w/w). Supplementations of oak sawdust at above 40%(w/w) of the waste and rice bran at 20%(w/w) of the sawdust allowed such a good mycelial growth as to be selected as a pertinent mixing ratio for fruiting medium. A fruiting yield on enokitake cultural waste supplemented by oak sawdust (at 40% of the waste, w/w) and rice bran (at 20% of the sawdust, w/w) was not inferior to that on oak sawdust supplemented by rice bran only (at 20% of the sawdust, w/w). These results indicated strongly the potentiality of enokitake cultural waste as raw materials for shiitake cultivating substrates.

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Studies on Submerged Culture and Mycelial Components of Naematoloma sublateritium Mycelia (개암버섯균의 액체배양과 균사체의 성분에 관한 연구)

  • Kang, An-Seok;Kang, Tae-Su;Cho, Soo-Muk;Yu, Seung-Hun
    • The Korean Journal of Mycology
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    • v.29 no.1
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    • pp.22-27
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    • 2001
  • This study was carried out to get the basic information for the submerged culture and analyze the biochemical components of Naematoloma sublateritium mycelia. The optimal temperature, pH, agitation speed and cultural time for the mycelial growth of Naematoloma sublateritium were $25^{\circ}C$, 5.5, 150rpm and 20 days, respectively. The proximate composition of mycelia was as follows; carbohydrate 55.8% (total sugar 48.7%), crude protein 22.4%, fat 4.1 % and ash 4.7% respectively. Among the free amino acid contents, phenylalanine, alanine and lysine were predominant component. The linoleic acid and palmitic acid were found to be the highest among the free fatty acids. The biopolymer extracts of mycelia was identified to be protein-bounded polysaccharide by color reaction and sepharose CL-4B gel chromatography.

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Fusarium Fruit Rot of Posthavest Oriental Melon (Cucumis melo L. var. makuwa Mak.) Caused by Fusarium spp. (Fusarium spp.에 의한 수확 후 참외 열매썩음병)

  • Kim, Jin-Won;Kim, Hyun-Jin
    • Research in Plant Disease
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    • v.10 no.4
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    • pp.260-267
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    • 2004
  • Fusarium spp. were isolated from the postharvest fruit rot of oriental melon fruits at commercial fruit markets in Korea during 2001 to 2003. The decayed fruits were covered with the fungal mycelia and eventually soft rotted. The disease started at the fruit stalk area, the calyx end of the fruit and skin of fruit. As the disease advanced, white to pinkish mycelia covered with the surface of decayed fruit. The cultural and morphological characteristic of Fusarium spp. were compared with descriptions of those reported previously, and identified as Fusarium equiseti, F. graminearum, F. moniliforme, F. proliferatum, F. sambucinum, and F. semitectum. Pathogenicity of the isolates was proved by artificial wound and unwound inoculation onto the healthy fruits. Two days after inoculation, aerial mycelia were noticed on the wound inocultion region of the fruit and developed soft rot symptoms. Although Fusarium spp. causing fruit rot disease in oriental melon have been reported in Korea, identification of the those species was not described. Therefore, this is the first report of Fusarium spp. causing postharvest fruit rot on oriental melon in Korea.

Characterization of D-Xylose Isomerase from Streptomyces albus (Stleptomyces albus의 D-Xylose Isomerase의 성질에 관하여)

  • 김영호;하영칠
    • Korean Journal of Microbiology
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    • v.16 no.2
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    • pp.47-61
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    • 1978
  • Strptomyces albus T-12 which ahd been isolated and identified in the laboratory, was selected for the studies on the cultural conditions on the production of D-xylose iosmerase and the enzymological characteristics using the partially purified enzyme. The best results in the enzyme production came from D-xylose medium than wheat bran. The divalent metla ions as $Co^{2+},\;Fe^{2+},\;Zn^{2+}\;and\;Cu^{2+}$ retard or inhibit the cell-growth at the early stages of mycelia propagations, and T-12 strain is especially sensitive to $Co^{2+}$. After 60 hours of shaking cultivation at $30^{\circ}C$ and 200 rpm, a maximum enzyme activitz, 0.49 enzyme units, was obtained. Cell-free enzyme obtained from mycelia heat-treated in the prescence of 0.5mM $Co^{2+}$, showed a 2.4-fold increase in specific than the enzyme from untreated mycelia. The specific activity of the purified enzyme through Sephadex G-150 columm showed 180 fold to the crude enzyme. The effective activators of the enzyme appeared to be $Mg^{2+}\;and\;Co^{2+}$ ions, and it exhibited the maximal enzyme activity showed at pH 7.0 and at tempersture around $80^{\circ}C$ when $Mg^{2+}\;and\;Co^{2+}$ ions were added. The enzyme isomerized D-glucose, D-xylose, D-ribose, L-arabinose, D-mannose, and L-rhamnose in the present of $Mg^{2+}\;and\;Co^{2+}$ ions as an activatiors. $Mg^{2+}\;and\;Co^{2+}$ ions were non-competitively bound at different allosterix sites of enzyme molecule. $Mg^{2+}(5mM)\;or\;Co^{2+}(1.0mM)$ protected against the thermal denaturations of the enzyme activities. The michelis constant(Km) and $V_{max}$ values of the emzyme for D-glucose and D-xylose were 0.52M, $2.12{\mu}moles/ml{\cdot}min.\;and\;0.28M,\;0.65moles/ml{\cdot}min.$, respectively.

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Occurrence Dollar Spot Caused by Sclerotinia homoeocarpa in Turfgrass of Golf Course in Korea (한국 골프장에서 Sclerotinia homoeocarpa에 의한 잔디동전마름병의 발생)

  • 심규열;민규영;신현동;이현주
    • Asian Journal of Turfgrass Science
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    • v.14 no.1
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    • pp.241-250
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    • 2000
  • In 1997, a new disease on creeping bentgrass and Kentucky bluegrass occurred in the green and fairway of a few golf courses in Korea. The disease spread gradually throughout the country and became a threat to turfgrass management. Symptoms of the disease consisted of small, circular, and sunken patches sized 3~5cm in diameter. The disease named as a dollar spot following its characteristic symptoms of circular blight and bleach on the green. The disease peaked two times in a year from April to June and from late August to October and white cottony mycelia of the causal fungus developed on diseased turfs in the early morning when the conditions were favored. A causal fungus was consistently isolated from the infected tufgrass and seven isolates originated from seven golf courses located in six provinces were selected for further study. The fungus produced abundant white aerial mycelia on PDA and turned to dark gray or light brown as it aged. Width of the mycelium was ca. $5~8\mu\textrm{m}$. While sclerotia were not readily formed on the medium, scattered small and dark colored stromata were developed on the surface. The fungus grew well on PDA between 5 to $30^{\circ}C$ and maximally around $25^{\circ}C$. Based on investigated mycological and cultural characteristics, the causal agent of dollar spot was identified as Sclerotinia homoeocarpa. The fungus showed strong pathogenicity to several turfs as creeping bentgrass, Kentucky bluegrass, perennial ryegrass, tall fescues, and zoysiagrass.

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Breadcrumb as a New Substrate for Trametes versicolor and Schizophyllum commune Submerged Cultivation

  • Ivanova, Tetiana S.;Bisko, Nina A.;Krupodorova, Tetiana A.;Barshteyn, Victor Yu.
    • Microbiology and Biotechnology Letters
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    • v.42 no.1
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    • pp.67-72
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    • 2014
  • A new substrate, breadcrumbs, was investigated for biomass accumulation, the pH of the cultural broth, the formation of primary metabolites such as the proteins and endopolysaccharides of Schizophyllum commune 1768 and Trametes versicolor 353, as well as its bioconversion efficiency. The results showed that S. commune gives more mycelial mass ($23.96{\pm}0.8g/l$) and in a shorter period (4 days) than T. versicolor ($15.76{\pm}0.5g/l$ in 5 days). The pH values changed from the initial 6.1 to 3.6 in S. commune cultural broth and to 4.4 in T. versicolor cultural broth. Maximal endopolysaccharide content in the mycelia of S. commune and T. versicolor were 7.13% and 6.42%, correspondingly. Crude protein content in S. commune mycelium was 18.83 % on the 4th day of cultivation, and 20.03%, in the mycelium of T. versicolor, on the 6th day of cultivation. Kinetic parameters for the quantitative estimation of cultivation efficiency were calculated for biomass, endopolysaccharide, and crude protein concentrations.

Cultural Characteristics for the Enhanced Mycelial Growth of Ramaria botrytis

  • Lee, Tae-Hee;Han, Yeong-Hwan
    • Mycobiology
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    • v.33 no.1
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    • pp.12-14
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    • 2005
  • The culture conditions for the enhanced mycelial growth of Ramaria botrytis was investigated. The optimal temperature and pH for the mycelial growth were $24^{\circ}C$ and 5.0, respectively. It was shown that starch was best of several carbon sources in Czapek-Dox medium as a minimal medium for the enhanced mycelial growth. Organic nitrogen sources were better than inorganic ones for mycelial growth. The appropriate vitamin and mineral salt were biotin and FeCl3, respectively. When this strain was cultured with $FeCl_3$ for 30 days, 19.23 g/l of dry mycelium of R. botrytis was obtained.