• The Korean Journal of Mycology
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• v.27 no.6 s.93
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• pp.373-377
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• 1999

This study was designed to develop functional drink from jujube extract through a simple submerged culture of three basidiomycetes species. The optimum Brix and pH of the jujube extract for culturing the Ganoderma lucidum appeared to be 5 Brix and pH 4. Ten days of culture period produced maximum mycelium. The optimum Brix and pH of the jujube extract for culturing the Coriolus versicolor appeared to be 5 Brix and pH 5. Ten days of culture period produced maximum mycelium. The optimum Brix and pH of the jujube extract for culturing the Phellinus igniarius appeared to be 3 Brix and pH 5. For the maximum mycelial production eighteen days of culture period was required for Phellinus igniarius. The antitumor activity of the polysaccharides extracted from the fermented drinks was demonstrated through the tumor cell line experiments. The $IC_{50}$ values of the jujube drinks fermented with Ganoderma lucidum and Phellinus igniarius against stomach cancer cell line appeared to be one fourth that of the jujube drink which was not fermented with basidiomycetes.

• Archives of Pharmacal Research
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• v.10 no.3
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• pp.158-164
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• 1987

To obtain a new strain of Ganoderma lucidum by protoplast fusion technique, its protoplast formation and regeneration were studied. Several factors affecting the protoplast formation and regeneration were investigated to find their optimum conditions. The mycelium was grown for four days on the cellophane membrane placed on G. Incidum complete medium (GCM). When various commercial lytic enzymes were examined for protoplast isolation, the combination of Novozym 234 and $\beta$glucuronidase was found to be effective. An osmotic stabilizer, 0.6 M sucrose in 20 mM phosphate buffer pH 5.8, gave the highest yield of protoplasts. Three-hour incubation in shaking incubator was most suitable for releasing protoplasts. To increase the protoplast yield, pretreatment with 2-mercaptoethanol was carried out. The regeneration frequency in GCM containing 0.6M MgSO$_4$ 7$H_2O$ was shown to be 0.66%.

• Microbiology and Biotechnology Letters
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• v.24 no.4
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• pp.459-464
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• 1996

Exo-polysaccharide (BWS) obtained from submerged cultivation of Ganoderma lucidum mycelium was fractionated. Antitumor activity of their fractions was investigated in comparison with the mycelial polysaccharide fractions. Eight kinds (BWS-DN, BWS-DA, BWS-DN-GI, BWS- DA-GI, MWS-DN, MWS-DA, MWS-DN-GI and MWS-DA-GI) of polysaccharide fractions were obtained by DEAE-cellulose ion exchange chromatography and Sepharose CL-4B gel chromatography from BWS and MWS, which were isolated from culture fiuid and mycelial cell, respectively. The anticomplementary activities (ITCH$_{50}$%) of the exo-polysaccharide fractions showing 15% to 30% were lower than those of mycelial polysaccharide fractions showing 15% to 70%. The acidic fractions of BWS-DA and BWS-DA-GI fractionated from BWS, showed the highest activity of 30%. In the MTT assay, BWS-DN and MWS against mouse leukemia L1210 exhibited high inhibition ratio of 86 and 89%, respectively at the concentration of 600 $\mu$g/ml. High inhibition ratio of 50% (IC$_{50}$) was achieved for BWS, BWS-DA and MWS-DA fractions against human colon adenocarcinoma COLO-205 and for BWS-DA, BWS-DN and MWS-DN fractions against human leukemia HL-60 at the concentra- tion of 300 $\mu$g/ml among the six polysaccharide fractions, respectively.

• Journal of Mushroom
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• v.2 no.2
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• pp.49-59
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• 2004

To establish a genetic relationships of collected Ganoderma strains, mycelium growth according to media and temperature, colony morphology, chlamydospore formation and fruitbody morphology were investigated. For the identification of optimal growth conditions of the strains, five different growth media and four different temperature were tested. GCM (Ganoderma complete medium) at $30^{\circ}C$ was the most effective for mycelial growth of 68 strains with more or less variation. The strains were divided into 28 groups based on their colony shapes, and most of them belong to CM3 or CM8 group. Chlamydospores were observed in the mycelia of 16 strains including ASI 7022 on microscope, but not in most G. lucidum domestic strains, which showed relatively lagging growth on $35^{\circ}C$ in mycelial growth experiment. These results were not similar to those of G. lucidum but those of G. tsugae imported from USA. The strains were cultivated on oak sawdust media to see their fruit body formation. Ninety-seven among 115 strains formed fruitbodies in sawdust cultivation. They showed two forms of fruitbodies, 89.7% of flat type or 10.3% of antler type, although these shapes can be affected by $CO_2$ concentrations. These results suggest that the native strains formerly considered to belong to G. lucidum have to be re-classified with further study.

• Journal of Applied Tourism Food and Beverage Management and Research
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• v.14 no.1
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• pp.47-58
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• 2003

Brown rice was used as material for solid-substrate cultivation of Ganoderma lucidum. The hydration time with cold water appeared to be 10 hours for brown rice, but the final water content was much less than optimum water content(65%). Hot water reduced the hydration time of brown rice, and the water content reached to 65% within 40 mins. From this result, hot water was better than cold water for the hydration of brown rice. We attempted to develop a practically applicable process by combining the soaking and sterilization. The water content of 65% appeared to be the best for the growth of the fungi and production of glucosamine related to the amount of mycelium. The content of free sugar increased far more in brown rice fermented with mycelium than in brown rice which was not fermented. Addition was most suitable 20% when add mushroom-rice to brown rice.

• Journal of Industrial Technology
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• v.25 no.B
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• pp.241-251
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• 2005

Exopolysaccharide (CBP) from submerged culture broth of Ganoderma lucidum mycelium and the water soluble (BWS) and water insoluble (BWI) fractions of CBP were prepared by gel filtration. Antitumor activity and effects on proliferation and differenciation of human cancer cells and mouse NIH 3T3 cells were studied. Cytotoxicity test of CBP, BWS and BWI fractions on human cancer cell lines was performed by using sulforhodamine B (SRB) assay. A549 (lung carcinoma), Colo320 DM and HSR (colon carcinoma), and NIH 3T3 cells were used. BWI fraction showed the strongest cytotoxicity (maximum 20% survival) to all human cells tested. However it did not induced apoptosis. Interestingly BWI fraction did not exert cytotoxic effect on NIH 3T3 cells at low concentration of cells ($5{\times}10^4$) but strong toxic effect at high concentration of cells($5{\times}10^5$) which showed transformed morphology. These results suggest that BWI may have cancer cell specific anticancer activity. However, BWI fraction did not effect the amount of pRb and c-myc protein, which implied that BWI fraction did not act at the early stage of signal transduction pathway. CBP fraction induced differenciation of human leukemic cell line, HL-60 cells suggesting the carcinogenesis prevention of normal cell and possible induction of normalization for cancer cell.

• Journal of Microbiology and Biotechnology
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• v.20 no.4
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• pp.844-851
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• 2010

Controlling the dissolved oxygen (DO) in the fed-batch culture of the medicinal mushroom Ganoderma lucidum led to a 2-fold increase of the maximum biomass productivity compared with uncontrolled DO conditions. By contrast, extracellular polysaccharide (EPS) production was two times higher under oxygen limitation (uncontrolled DO) than under increased oxygen availability (controlled DO). Morphologically, dispersed mycelium was predominant under controlled DO conditions, with highly branched hyphae, consistent with the enhanced culture growth noted under these conditions, whereas in the uncontrolled DO process mycelial clumps were the most common morphology throughout the culture. However, in both cultures, clamp connections were found. This is an exciting new finding, which widens the applicability of this basidiomycete in submerged fermentation. In rheological terms, broths demonstrated shear-thinning behavior with a yield stress under both DO conditions. The flow curves were best described by the Herschel-Bulkley model: flow index down to 0.6 and consistency coefficient up to 0.2 and 0.6 Pa $s^n$ in uncontrolled and controlled cultures DO, respectively. The pseudoplastic behavior was entirely due to the fungal biomass, and not to the presence of EPS (rheological analysis of the filtered broth showed Newtonian behavior). It is clear from this study that dissolved oxygen tension is a critical process parameter that distinctly influences G. lucidum morphology and rheology, affecting the overall performance of the process. This study contributes to an improved understanding of the process physiology of submerged fermentation of G. lucidum.

• The Korean Journal of Mycology
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• v.27 no.1 s.88
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• pp.76-81
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• 1999

Exo-polysaccharide obtained from the submerged cultivation of Ganoderma lucidum mycelium was fractionated. The structural analysis of the acidic exo-polysaccharide fraction (BWS-DA-GI), showing high antitumor activity, was carried out and compared to the mycelial acidic fraction (MWS-DA-GI). The major sugar constituents of the fraction of BWS-DA-GI were glucose, galactose and mannose in the molar ratio of 2.5 : 2.1 : 2.5. The minor components in this fraction were xylose and fucose. While the major sugar constituents of the mycelial acidic fraction of MWS-DA-GI were galactose, fucose, mannose and glucose. The trace components in this fraction was xylose. From the results of periodate oxidation, Smith degradation, affinity chromatography and methylation analysis, the chemical structures of the two fractions, BWS-DA-GI and MWS-DA-GI were both determined as ${\beta}-1,3$ glucans. It was also estimated that BWS-DA-GI had a $1{\rightarrow}6$ glucosidic linkage and MWS DA-GI had $1{\rightarrow}4$ and $1{\rightarrow}6$ glucosidic linkages. The molecular weights of these fractions, MWS-DA-GI and MWSDA-GI were estimated as $1.2{\times}10^6\;and\;1.0{\times}10^6$ dalton, respectively.

• Korean Journal of Food Science and Technology
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• v.43 no.5
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• pp.633-640
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• 2011

After Ganoderma lucidum was cultured in mushroom complete medium (MCM) supplemented with ginseng extract (GE), crude polysaccharide (GL-GE-CP) was fractionated from mycelium. Among GL-GE-CP from mycelium in MCM supplemented with 5, 10, and 15% GE (v/v ratio of MCM to GE), GL-GE-15-CP (15% GE) most significantly enhanced macrophage stimulation and intestinal immune system modulating activity compared with GL-CP in MCM without GE. When GL-GE-15-CP was further fractionated on DEAE-Sepharose CL-6B, GL-GE-15-CP-II displayed more potent activity than subfractions from GL-CP on macrophage stimulation, interleukin-12 production, and intestinal immune system modulation (1.75-, 5.68-, and 1.76-fold, respectively). Anti-metastasis effect against colon 26-M3.1 carcinoma cells was also enhanced by GL-GE-15-CP-II (72.8% inhibition). In addition, GL-GE-15-CP-II contained neutral sugar (83.00%) and uronic acid (9.11%), and consisted of Ara, Man, Gal and Glc (molar ratio of 0.39:0.50:0.75:1.00). Furthermore, GE supplementation helped to enhance the immunomodulation in G. lucidum, and it is assumed that neutral polysaccharides play an important role.

• The Korean Journal of Mycology
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• v.18 no.3
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• pp.137-144
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• 1990

To examine effects on complement and reticuloendothelial system, alkali extract was isolated from cultured mycelium of Ganoderma lucidum IY107. It was shown to strongly activate both classical and alternative pathways of complement as compared with krestin. Activated complement C3, 3rd peak, was observed by crossed immunoelectrophoresis. It was also shown to activate reticuloendotherial system of ICR mice in the carbon clearance test and to increase hemolytic plaque forming cells of the spleen. Carbohydrate and protein contents of the alkali extract were 10% and 49%, respectively. The carbohydrate consisted of four monosaccharides and the protein contained 16 amino acids.