• Title/Summary/Keyword: mycelial cells

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FUNGAL EXTRACELLULAR POLYSACCHARIDES INVOLVED IN RECYCLING OF METABOLITES AND OSMOTOLERANCE OF PENICILLIUM FELLUTANUM : APPLICATION OF $^{13}$ C-NMR SPECTROSCOPY FOR THE STUDY ON FUNGAL PHYSIOLOGY AND METABOLISM

  • Park, Yong-Il;Gander, John.-E.
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2000.04a
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    • pp.208-213
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    • 2000
  • Penicillium fellutanum produces a phosphorylated, choline-containing extracellular peptido-polysaccharide, peptidophosphogalactomannan (pPxGM) (8). The $\^$13/C-methyl labeled pPxGM ([methyl-$\^$13/C]pPxGM) was prepared from the cultures supplemented with L-[methyl-$\^$13/C]methionine or [2-$\^$13/C]glycine and was used as a probe to monitor the fate of phosphocholine in this polymer. Addition of purified [methyl-$\^$l3/C]pPxGM to growing cultures in low phosphate medium resulted in the disappearance of [methyl-$\^$13/C]phosphocholine and -N,N'-dimethyl-phosphoethanolamine from the added [methyl-$\^$13/C]pPxGM. Two $\^$l3/C-methyl-enriched cytoplasmic solutes, choline-O-sulfate and glycine betaine, were found in mycelial extracts, suggesting that phosphocholine-containing extracellular pPxGM of P.fellutanum is a precursor of intracellular choline-O-sulfate and glycine betaine and thus of phosphatydilcholine (l0). $\^$13/C-Methyl-labeled cells grown in 3 M NaCl-containing medium showed 2.6- and 22-fold more accumulation of $\^$13/C-methyl labeled choline-O-sulfate and glycine betaine, respectively, originated from the extracellular [$\^$13/C-methyl]pPxGM than those grown without added NaCl. The results suggest that, in addition to glycerol and erythritol, glycine betaine and choline-O-sulfate and thus choline are also osmoprotectants and hence that pPxGM is involved in osmotolerance of this fungus (11). Taken collectively, the $\^$l3/C- and $\^$31/P-NMR analyses of cytosolic solute pools and structural modulation of extracellular pPxGM corresponding to environmental stimuli in P. fellutanum, provided evidence that pPxGM is involved in cellular choline metabolism, osmotolerance, and recycling of metabolites.

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Antagonistic activity of Streptomyces apecies against Fusarium solani causing ginseng root rot (인삼뿌리 썩음 병균 Fusarium solane에 대한 Streptomyces species의 길한작용)

  • 정영륜;오승환;정후섭
    • Korean Journal of Microbiology
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    • v.27 no.1
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    • pp.56-62
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    • 1989
  • Antagonistic effects of Streptomyces species aganinst Fusarium solani causing ginseng root rot were investigated in terms of chitinase activity and growth inhibition in vitro. Among 131 isolates of streptomycetes obtained from ginseng cultivating soil, 9 isolates producing large clear zone around the colony on a chitin agar medium were selected for further study. All 9 isolates produced chitinase in a range from 0.10 to 0.38 U lysing cells of F. solani and inhibited germination of the conidia. In the ten-fold condentrated culture filtrate of S. alboniger ST59 and S. roseolilacinus ST129, the number of conidia of F. solane was reduced to about 20% of original count within 14 days. When S. alboniger ST59 and F. solani were grown simultaneously in the mineral saly medium, chitinase activity increased with incubation period, whereas mycelial volume of F. solani decreased. In a chitin added mineral salt medium, chitinase activity increased during the first four days and maintained steady level until the 8th day, and increased thereafter. S. alboniger ST59 lysed mycelia, conidia and even chlamydospores of F. solani. It is probable that the antagonistic activity of this streptomycete against F. solani is the lysis of fungal cell wall by streptomycete producing chitinase affected by antifungal substances.

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Pyridoxatin, an Inhibitor of Gelatinase A with Cytotoxic Activity

  • Lee, Ho-Jae;Chung, Myung-Chul;Lee, Choong-Hwan;Chun, Hyo-Kon;Kim, Hwan-Mook;Kho, Yung-Hee
    • Journal of Microbiology and Biotechnology
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    • v.6 no.6
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    • pp.445-450
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    • 1996
  • Gelatinase A is a member of the matrix metalloproteinases that play an important role in cancer invasion and metastasis. In the course of screening gelatinase A inhibitors from microbial sources, a fungal strain PT-262 showed a strong inhibitory activity. The strain was identified as Chaunopycnis alba on the basis of its morphological characteristics. The inhibitor was isolated from acetone extract of mycelial cake by sequential chromatographies on MCI-gel, Sephadex LH-20, and a reverse-phase HPLC column. The purified inhibitor was identified as pyridoxatin by its physico-chemical properties and spectroscopic analysis. Pyridoxatin is not a peptide analog and has cyclic hydroxamic acid moiety. It inhibited activated gelatinase A with an $IC_{50}$ value of 15.2 ${\mu}M$ using fluorescent synthetic peptide. It also had a strong cytotoxicity against human cancer cell lines in vitro. Furthermore, this compound inhibited DNA synthesis with an $IC_{50}$ value of 2.92 ${\mu}M$ in PC-3 prostate cancer cells by [$^3H$]thymidine incorporation assay.

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Evaluation of the Synergistic Effect of Mixed Cultures of White-Rot Fungus Pleurotus ostreatus and Biosurfactant-Producing Bacteria on DDT Biodegradation

  • Purnomo, Adi Setyo;Ashari, Khoirul;Hermansyah, Farizha Triyogi
    • Journal of Microbiology and Biotechnology
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    • v.27 no.7
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    • pp.1306-1315
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    • 2017
  • DDT (1,1,1-trichloro-2,2-bis(4-chlorophenyl) ethane) is one of the organic synthetic pesticides that has many negative effects for human health and the environment. The purpose of this study was to investigate the synergistic effect of mixed cutures of white-rot fungus, Pleurotus ostreatus, and biosurfactant-producing bacteria, Pseudomonas aeruginosa and Bacillus subtilis, on DDT biodegradation. Bacteria were added into the P. ostreatus culture (mycelial wet weight on average by 8.53 g) in concentrations of 1, 3, 5, and 10 ml ($1ml{\approx}1.25{\times}10^9$ bacteria cells/ml culture). DDT was degraded to approximately 19% by P. ostreatus during the 7-day incubation period. The principal result of this study was that the addition of 3 ml of P. aeruginosa into P. ostreatus culture gave the highest DDT degradation rate (approximately 86%) during the 7-day incubation period. This mixed culture combination of the fungus and bacteria also gave the best ratio of optimization of 1.91. DDD (1,1-dichloro-2,2-bis(4-chlorophenyl) ethane), DDE (1,1-dichloro-2,2-bis(4-chlorophenyl) ethylene), and DDMU (1-chloro-2,2-bis(4-chlorophenyl) ethylene) were detected as metabolic products from the DDT degradation by P. ostreatus and P. aeruginosa. The results of this study indicate that P. aeruginosa has a synergistic relationship with P. ostreatus and can be used to optimize the degradation of DDT by P. ostreatus.

Differential Expression of Laccase Genes in Pleurotus ostreatus and Biochemical Characterization of Laccase Isozymes Produced in Pichia pastoris

  • Park, Minsa;Kim, Minseek;Kim, Sinil;Ha, Byeongsuk;Ro, Hyeon-Su
    • Mycobiology
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    • v.43 no.3
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    • pp.280-287
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    • 2015
  • In this study, transcriptome analysis of twelve laccase genes in Pleurotus ostreatus revealed that their expression was differentially regulated at different developmental stages. Lacc5 and Lacc12 were specifically expressed in fruiting bodies and primordia, respectively, whereas Lacc6 was expressed at all developmental stages. Lacc1 and Lacc3 were specific to the mycelial stage in solid medium. In order to investigate their biochemical characteristics, these laccases were heterologously expressed in Pichia pastoris using the pPICHOLI-2 expression vector. Expression of the laccases was facilitated by intermittent addition of methanol as an inducer and sole carbon source, in order to reduce the toxic effects associated with high methanol concentration. The highest expression was observed when the recombinant yeast cells were grown for 5 days at $15^{\circ}C$ with intermittent addition of 1% methanol at a 12-hr interval. Investigation of enzyme kinetics using 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) as a substrate revealed that the primordium-specific laccase Lacc12 was 5.4-fold less active than Lacc6 at low substrate concentration with respect to ABTS oxidation activity. The optimal pH and temperature of Lacc12 were 0.5 pH units and $5^{\circ}C$higher than those of Lacc6. Lacc12 showed maximal activity at pH 3.5 and $50^{\circ}C$, which may reflect the physiological conditions at the primordiation stage.

Studies on Constituents of the Higher Fungi of Korea(XLI) -An Antitumor Fraction from the Culture Filtrate of Lentinus edodes DMC7- (한국산(韓國産) 고등(高等) 균류(菌類)의 성분(成分) 연구(硏究)(제41보)(第41報) -Lentinus edodes DMC7 균주(菌株)의 배양(培養) 여액(濾液)의 항암(抗癌) 성분(成分)-)

  • Chung, Kyeong-Soo;Choi, Eung-Chil;Kim, Byong-Kak
    • The Korean Journal of Mycology
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    • v.12 no.4
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    • pp.129-132
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    • 1984
  • To find antitumor constituents in Korean basidiomycetes, the mycelia of Lentinus edodes DMC7, which had shown a good mycelial growth in shakeflasks, were cultured at $27^{\circ}C$ on an orbital shaking incubator at 180 rev/min for 12 days. The medium was composed of glucose (50g/l), yeast extract (9g/l), peptone (9g/l), and seven inorganic salts. A water soluble macromolecular fraction, LF-3, was obtained from the culure filtrate by fractionation with ethanol and dialysis using a Visking tube. When LF-3 was administered i.p. at 50mg/kg/day once daily for 10 consecutive days to female ICR mice which were implanted s.c. with sarcoma 180 $(10^6\;cells/mouse)$, it exerted a highly significant antitumor activity, with the tumor inhibition ratio of 53. 1%.

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Effect of Ganoderma lucidum Solid-state Fermented Salix gracilistyla Extract on Type I Procollagen Biosynthesis in HDFn Cells (불로초 균사배양 갯버들 추출물이 인간 피부 섬유아세포의 제1형 프로콜라겐 생성에 미치는 영향)

  • Jeong, Yong-Un;Park, Young-Jin
    • The Korean Journal of Mycology
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    • v.47 no.2
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    • pp.153-163
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    • 2019
  • This study was carried out to investigate the feasibility of Salix gracilistyla production for cosmetic use through mycelial fermentation. The efficacy of this method was confirmed by fermentation using the mycelia of Ganoderma lucidum (a representative medicinal mushroom). Total polyphenol and flavonoid content and DPPH radical scavenging activity of S. gracilistyla extract (SGE) were found to be higher than those of G. lucidum fermented S. gracilistyla extract (GLSGE). GLSGE had relatively lower collagenase activity than SGE. However, GLSGE increased HDFn cell viability more potently than SGE, and increased the biosynthesis of type I procollagen. Thus, GLSGE could be used as an anti-aging cosmetic active ingredient. These results indicate that extract fermentation using G. lucidum mycelia can effectively enhance some beneficial effects of functional materials.

Production of Nitric Oxide in Raw 264.7 Macrophages treated with Ganoderan, the ${\beta}-Glucan$ of Ganoderma lucidum (영지의 균사체성 ${\beta}-glucan$에 의한 Raw 264.7 대식세포의 Nitric Oxide생성)

  • Han, Man-Deuk;Lee, Eun-Sook;Kim, Young-Kweon;Lee, June-Woo;Jeong, Hoon;Yoon, Kyung-Ha
    • The Korean Journal of Mycology
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    • v.26 no.2 s.85
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    • pp.246-255
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    • 1998
  • Ganoderan (GAN), an immunomodulating ${\beta}-glucan$ of G. lucidum, induces potent antitumor immunity in tumor-bearing mice. This study was set up to elucidate the ability of macrophage activation of GANs. GAN-treated Raw 264.7 macrophages showed enhanced production of nitric oxide (NO). The ability of GANs to produce NO was based on differences in chemical composition of GANs obtained from the mycelium on various carbon sources and mycelial fractionation. The highest NO production was observed in CW-AS-WS polysaccharide which was extracted from the mycelial wall. GAN-treated Raw 264.7 cells gave a 2-to 5-fold (24 hr) formation of NO levels compared with those treated with medium only. Partial removal of the protein in the extracellular GAN by TCA treatment did appreciably reduce its capacity to secrete NO. The mixture effect of GAN and LPS increased the nitric oxide secretion from RAW 264.7. The cell proliferation of GAN-treated Raw 264.7 cell tines inhibited as compared with its control. Of the culture supernatant of macrophage activated by GAN, the percentage of cytotoxicity against mouse leukemia L1210 cells was slightly dependent on the amount of NO in the culture supernatants of the activated-macrophages. These results indicate that the ${\beta}-glucan-related$ polysaccharides of the higher fungus activate macrophage and release nitric oxide. It also suggests that murine macrophages possess certain receptors for ${\beta}-anomeric$ glucans and play a critical role of ${\beta}-glucan-related$ tumor killing mechanism.

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Enhancement of Anticarcinogenic Potentials of Submerged-Liquid Culture of Agaricus blazei Murill on Mouse Ascites Cancer by Rice Hull (왕겨에 의한 신령버섯균사체 액체배양액의 생쥐 항복수암성 증가)

  • Kim, Young-S.;Jang, Wook-J.;Rakib, A.;Kwon, Jung-M.;Ahn, Chae-R.;Kim, So-Y.;Cho, Yong-U.;Ha, Young-K.;Kim, Jeong-O.;Ha, Yeong-L.
    • Journal of Life Science
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    • v.20 no.9
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    • pp.1402-1408
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    • 2010
  • The effects of rice hull (RH) powder on the anticarcinogenic activity of submerged-liquid cultures of Agaricus blazei Murill (AB) were assessed for mouse ascites cancers induced by mouse Sarcoma S-180 (S-180) cancer cells. Optimal growth of AB mycelia in the basal liquid culture medium, containing soybean meal, was achieved by culturing at $25^{\circ}C$ for 5 days, when evaluated by $\beta$-glucan content, Brix, and mycelial weight, relative to other culture conditions. Hot-water extract (HWE) of the submergedliquid culture of AB mycelia grown at $25^{\circ}C$ for 5 days exhibited a stronger anticarcinogenic activity, relative to HWE from other culture conditions. No such effects were obtained from AB mycelial cultures by alternative temperature-controlling cultures. Both cytotoxicity for S-180 cells and anticarcinogenic potentials for mouse ascites cancer of the HWE from AB mycelia grown in the basal medium containing 1% RH powder for 5 days at $25^{\circ}C$ were significantly (p<0.05) enhanced, relative to HWE from the AB mycelia culture of the basal medium without RH powder. These results indicate that HWE of submerged-liquid culture of AB mycelia, incubated in media containing 1% RH powder at $25^{\circ}C$ for 5 days, enhanced anticarcinogenic activity against S-180 cell-induced mouse ascites cancer, and suggest that RH powder is an excellent ingredient for the improvement of the anticarcinogenic potentials of the submerged-liquid culture of mushroom mycelia.

Occurrence of Powdery Mildew on Mung bean(Vigna radiatus L.) Caused by Sphaerotheca phaseoli (Sphaerotheca phaseoli에 의한 녹두 흰가루병 발생)

  • Lee, Sang-Yeob;Hwang, Soon-Jin;Lee, Snag-Bum
    • Research in Plant Disease
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    • v.8 no.3
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    • pp.166-170
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    • 2002
  • Powdery mildew of mug bean was found in Suwon on September 2000. Many white and powdery mycelial colonies appeared on leaves of the plants. Conidia with fibrosin bodies were ellipsoid to elongate-ellipsoid, 25~38$\times$16~22 ${\mu}{\textrm}{m}$ in size and farmed in chains. Conidiophores were erect on superfical mycelium, 67~100 ${\mu}{\textrm}{m}$ in length. Foot cells were cylindric with 37~65$\times$10~11 ${\mu}{\textrm}{m}$ in size. Appressoria were indisdinct. Germ tubes from conidia were simple, not branched. Dark brown cleistothecia were observed on the leaves of mung bean. Cleisto-thecia were globose or subglobose, 85~1130 ${\mu}{\textrm}{m}$ in size. Wall cells were irregularly polygonal with 15~45$\times$7~26 ${\mu}{\textrm}{m}$. Appendages were mycelioid, 6-11 per ascocarp, coloured, throught when mature or paler upwards, 1~7 septa, 192~223$\times$5~7 ${\mu}{\textrm}{m}$ in size. A single sun had in a cleistothecium and was ovoid or subglobose with 62~95$\times$50~67 ${\mu}{\textrm}{m}$ in size. Ascusspore had 8 spores in an ascus and rarely developed, mostly imatuna ellipsoid-ovoid with 16~24$\times$12~16 ${\mu}{\textrm}{m}$. On the basis of morphological characters of the conidial stage and teleo-morph, the fungus was identified as Sphaerotheca phaseoli.