• The Korean Journal of Mycology
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• v.23 no.4 s.75
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• pp.285-297
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• 1995

Ganoderan, an immunomodulating ${\beta}-glucan$ of G. lucidum, induces potent antitumor immunity in tumor-bearing mice. The present study was set up to elucidate the chemical composition and bioactivities of ganoderan obtained from the mycelial fractionation of G. lucidum IY009. Ganoderan was isolated and purified from its extracellular, cell wall and cytoplasmic sources. These ganoderans were composed mainly of glucose. The cell wall-alkali soluble-water soluble fraction (CW-AS-WS) showed the highest antitumor activity (inhibition rate of 94%) in sarcoma-bearing mice and 37% of anticomplementary activity. The CW-AS-WS fraction was found to be approximately average 20,000 dalton in aq. 0.3N NaOH solution and composed of 88% carbohydrate and 4% protein. The carbohydrate of the CW-AS-WS was composed of 74% glucose. These results indicate that the ganoderans extracted from the mycelial fractionations of G. lucidum IY009 had different chemical characteristics and showed different potentiality in antitumor and anticomplementary activity.

• The Plant Pathology Journal
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• v.23 no.3
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• pp.180-186
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• 2007

The biocontrol agent, Serratia plymuthica A21-4, has been developed for controlling pepper Phytophthora blight. Serratia plymuthica A21-4 strongly inhibits the mycelial growth, zoospore formation, and cyst germination of Phytophthora capsici in vitro. The application of a cell suspension of strain A21-4 to pepper plants in pot experiments and in greenhouse successfully controlled the disease. The bacteria produced a potent antifungal substance which was a key factor in the suppression of Phytophthora capsici. The most active chemical com-pound was isolated and purified by antifungal activity-guided fractionation. The chemical structure was identified as a chlorinated macrolide $(C_{23}H_{31}O_8Cl)$ by spectroscopic (UV, IR, MS, and NMR) data, and was named macrocyclic lactone A21-4. The active compound significantly inhibited the formation of zoosporangia and zoospore and germination of cyst of P. capsici at concentrations lower than $0.0625{\mu}g/ml$. The effective concentrations of the macrocyclic lactone A21-4 for $ED_{50}$ of mycelial growth inhibition were $0.25{\mu}g/ml,\;0.25{\mu}g/ml,\;0.30{\mu}g/ml \;and\;0.75{\mu}g/ml$ against P. capsici, Pythium ultimum, Sclerotinia sclerotiorum and Botrytis cinerea, respectively.

• Microbiology and Biotechnology Letters
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• v.28 no.4
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• pp.209-213
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• 2000

The cultured mycelia of Ganoderma lucidum were extracted and the extract was separated into six fractions by organic solvent fractionation. The antihepatotoxic activity of all the fractions was evaluated by measuring activities of glutamic pyruvic transaminase (GPT) and glutamic oxaloacetic transaminase (GOT). Among the fractions tested, the high-polarity fractions such as aqueous and n-butanol fractions significantly reduced activities of GPT and GOT in CCl4- and galactosamine-intoxicated rat primary hepatocytes. When intracellular synthetic activities were measured by pulsing the rate primary cultured hepatocytes with [3H]-uridine and [3H]-leucine, activities of DNA, RNA and protein. When direct toxicities of the fractions were measured against human hepatoma(SK-Hep-1), the non-polarity fractions such as n-butanol and ethyl acetate fractions showed potent direct cytotoxicities even at the concentration of 1 $\mu\textrm{g}$/ml. These data showed that Ganoderma lucidum has hepatoprotective and hepatotoxic recovery principles in its mycelia.

• Korean Journal of Microbiology
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• v.19 no.3
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• pp.142-152
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• 1981

One hundred and thirteen healed pulmonary tuberculosis patients and 11 patients with other underlying diseases were studied for evidence of pulmonary fungal infection because of persisting hemoptysis or chronic cough. Rediological, mycological and serological investigations revealed that 54 out of 124 patients were evidently infected with one or more species of fungi. A. fumigatus was isolated from 4 out of 70 patients whose sera did not react with antigens from this fungus, while it was isolated from 43 out of 47 serological reactors to this fungus. Chest radiography showed a distinct fungus ball in a cyst of one patient and in a preformed cavity in the lung of 17 healed tuberculosis patients and two other patients. The latter two patients were infected with A.flavus. Two patients, who were under the long period of immunosuppressive therapy, apparently succumbed to invasive aspergillosia due to A.fumigatus. A single or dual infection with A. flavus, A. nidulans, A.nidulans var. latus, C. albicans, and P. boydii were noticed in some patients without mycetomal shadow on chest radiographs. Young mycelial extract (ME) of A.fumigatus detected antibody in 95.8 percent of the sera from patients infected with this fungus, while it was isolated from 43 out of 47 serological reactors to this fungus. Chest radiography showed a distinct fungus ball in a cyst of one patient and in a performed cavity in the lung of 17 healed tuberculosis patients and two other patients. The latter two patients were infected with A. flavus. Two patients, who were under the long period of immunosuppressive therapy, apparently succumbed to invasive aspergillosis due to A.fumigatus. A single or dual infection with A. flavus, A. nidulans, A. niduans var. latus, C. albicans, and P. boydii were noticed in some patients without mycetomal shadow on chest radiographs. Young mycelial extract (ME) of A.fumigatus detected antibody in 95.8 percent of the sera from patients infected with this fungus, while the commercial culture filtrate antigen (GL) yielded 78.7 per cent positive result. Culture filtrate antigen, however, was comparable with ME. There was no single antigen with which all the serum specimens reacted. Fractionation of ME resulted in a loss of some activity although it excluded substances that reacted with C-reactive protein in a loss of some activity although it excluded substances that reacted with C-reactive protein. Most reactive and specific precipitinogens distributed in the fraction (FB) which was precipitable at 75 percent saturation with ammonium sulfate and eluted in a second peak in order from gel-filtration and which contained mostly proteinic components. Glycoproteins or polysaccharides rich fractions (FA and ASI) were relatively less effective in detecting antibody. Demonstration of antibody in the serum from patients using a battery of fungal antigens and of etiologically related fungi from clinical specimens are very useful laboratory procedures for the diagnosis of pulmonary fungal infection which is a common complication of tuberculosis.

• 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%.

• Food Science and Preservation
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• v.6 no.4
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• pp.500-505
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• 1999

The investigation was carried out to identify the active constituent in yeast extract for fruit body formation of Lentinula edodes. The result suggests that free thiamin which is hewn as the active substance for the fruiting of L edodes, was detected but thiamin mono, di, three phosphates were not detected in the yeast extract produced by Difco Co.. Therefore, the thiamin content of the yeast extract was determined, the yeast extract was fractionated to five portion by the post-column fluorescence method. The content of thiamin in yeast extract( 1g) was 0.436mg as thiamin hydrochloride. It was found that 76% of the total thiamin(0.332mg) was contained in fraction II. About 20% of the total thiamin(0.087mg) was present in fraction I, but not in fractions III, IV and V. In accordance with the contents of thiamin in the fractions, the fruit body formation was the highest by the treatment of fraction II(100%) and followed by fraction I (60%), V(50%), III(30%). Thiamin did not influence for the vegetative mycelial growth of L. edodes, but be used for fruit body formation.

• Journal of Korea Foresty Energy
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• v.17 no.1
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• pp.56-70
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• 1998

This study was carried out to investigate the structural characteristics of kraft lignin and the wood degrading characteristics, the productivity of ligninolytic enzymes and the enzymatic degradation of kraft lignin by white-rot fungi. To purify kraft lignin, precipitation of kraft pulping black liquors of pitch pine meal was done by titration with lN $H_{2}SO_{4}$ reaching to pH 2, and isolation of the precipitates done by centrifugation. The isolated precipitates from pitch pine were redissloved in lN NaOH, reprecipitated by titration with lN $H_{2}SO_{4}$, washed with deionized water, and kept ofr analysis after freeze drying. Fractionation of the precipitates in solution by successive extraction with $CH_{2}Cl_{2}$ and MeOH, and the fractionates were named SwKL, SwKL I, SwKL II, and SwKL III for pitch pine kraft lignin. The more molecular weights of kraft lignin increased, the less phenolic hydroxyl groups and the more aliphatic hydroxyl groups. Because as the molecular weights increased, the ratio of etherified guaiayl/syringyl(G/S ratio) and the percentage were increased. The spectra obtained by 13C NMR and FTIR assigned by comparing the chemical shifts of various signals with shifts of signals from autherized ones reported. The optimal growth temperature and pH of white-rot fungi in medium were $28^{\circ}C$ and 4.5-5.0, respectively. Especially, in temperature and pH range, and mycelial growth, the best white-rot fungus selected was Phanerochaete chrysosporium for biodegradation. For the degradation pathways, the ligninolytic fungus jcultivated with stationary culture using medium of 1% kraft lignin as a substrate for 3 weeks at $28^{\circ}C$. The weight loss of pitch pine kraft lignin was 15.8%. The degraded products extracted successively methoanol, 90% dioxane and diethyl ether. The ether solubles were analyzed by HPLC. Kraft lignin degradation was initiated in $\beta$-O-4 bonds of lignin by the laccase from Phanerochaete chrysosporium and the degraded compounds were produced from the cleavage of $C\alpha$-$C\beta$ linkages at the side chains by oxidation process. After $C\alpha$-$C\beta$ cleavage, $C\alpha$-Carbon was oxidized and changed into aldehyde and acidic compounds such as syringic acid, syringic aldehyde and vanilline. And the other compound as quinonemethide, coumarin, was analyzed. The structural characteristics of kraft lignin were composed of guaiacyl group substituted functional OHs, methoxyl, and carbonyl at C-3, -4, and -5 and these groups were combinated with $\alpha$ aryl ether, $\beta$ aryl ether and biphenyl. Kraft lignin degradation pathways by Phanerochaete chrysosporium were initially accomplished cleavage of $C\alpha$-$C\beta$ linkages and $C\alpha$ oxidation at the propyl side chains and finally cleavage of aromatic ring and oxidation of OHs.

• 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.