• KSBB Journal
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• v.25 no.2
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• pp.155-166
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• 2010

Studies on the production of cell-wall bound innerpolysaccharides (IPS) (soluble ${\beta}$-D-glucan) have been performed by use of suspended myelial cultures of Inonotus obliquus. This product has promising potentials as an effective antidiabetic as well as an immunostimulating agents. As a first step to enhanced production of IPS, Intensive strain improvement programs were carried out by obtaining a large amounts of protoplasts for the isolation of single cell colonies. Rapid and large screening of high-yielding producers was possible because about fivefold higher amount of protoplasts ($2.3{\times}10^6$ protoplasts/mL) could be recovered with relatively high regeneration rates of $10^{-2}{\sim}10^{-3}$ by applying a modified filtration method, as compared to the previously used trapping method. A basic protocol necessary for UV-mutation of the protoplasts was also developed, resulting in several overproducing variants with good fermentation properties. Since the amount of IPS extracted from the mycelial cell walls of I. obliquus turned out to be almost constant per g DCW, increase in cell mass was considered the most important factor for the enhancement in IPS production. Therefore, attempts were made to screen mutant cells showing rapid mycelial growth rate in the final suspended cultures. Notably, the mutant strains showing an active cellgrowth in the preceding solid growth cultures were observed to produce higher amount of IPS in the suspended fermentations as well. A striking mutant, OBLQ756-15-5 strain, obtained from the survivors of a harsh UV-treated condition (97% death rate) was found to stably produce as high cell mass as 22 g DCW/L in the final fermentations. Currently, this strain is being tested for development of a scaled-up fermentation process for mass production of IPS.

• Korean journal of applied entomology
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• v.12 no.2
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• pp.71-78
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• 1973

The study was performed to clear the effects of thiamine, biotin, nicotinic acid, pyridoxine, inositol, deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) on the mycelial growth and the sclerotial production of Sclerotium rolfsii Sacc. isolated from Magnolia kobus. The results are abstracted as follows: 1. Tested fungus was thiamine- deficient and required thiamine 20r/l for maximum growth of mycelia. At higher concentrations than thiamine 20r/l, however, mycelial growth was decreased with increasing the concentrations and was inhibited little less than that of thiamine-free control at 150r/l. 2. The effecfivenesses of the nitrogen sources on the mycelial growth under the thiamine presence were recognized in order of $NH_4NO_3>(NH_4)_2SO_4 >asparagine> KNO_3$, and on the sclerotial production were $KNO_3>NH_4NO_3>asparagine>(NH_4)_2SO_4$. The optimum concentrations of thiamine were about 12r/1 in $KNO_3$, about 16r/1 in asparagine on the growth of mycelia, and were about 8r/l in $KNO_3\;and\; NH_4NO_3,\; 16r/1$ in asparagine on the production of sclerotia. 3. After the organism began to grow, the pH value of cultral filtrate was rapidly dropped down to about 3.5. Hereafter it was slowly fallen down as the growth amount was increased, but was not depreciated below pH 2.2. 4. Nicotinic acid was not effective individually on the mycelial growth and the sclerotial formation of tested fungus without thiamine, but slight effect of it was recognized with thiamine 10r/l, even though maximum growth was shown at 7-10mg/1. Beyond that concentration, however, mycelial growth was rather depressed. 5. When ammonium sulphate or asparagine as the nitrogen sources was used, pyridoxine, biotin and inositol had not any effectivenesses on the mycelial growth and the sclerotial production of examined fungus. 6. In the concentrations of thiamine, biotin, pyridoxine and inositol, as long as thiamine was not added in those, their correlating effects on the growth of the organism were not observed at all. Equivalent or more effects on the mycelial growth were recognized in combinations of thiamin + pyridoxine, thiamine + inositol, thiamine + biotin + pyridoxine, and thiamine + biotin + pyridoxine + inositol compared with thiamino alone, and in combinations of thiamine + biotin and thiamine + biotin + inositol, mycelial growth was inhibited rather than that of thiamine alone. Sclerotial production of those combinations was increased more than that of thiamine alone in dry weight. 7, The little effects of DNA and RNA on the mycelial growth of the organism were recognized compared with the control(DNA-and RNA-free), and RNA was more effective than DNA. Maximum growth of mycelia was observed at RNA 2-6mg/1 and DNA 6mg/l. No effectivenesses on the sclerotial production were recognized in the RNA and DNA. 8. Mycelial growth of the organism was increased with increasing the concentrations of the RNA and the thiamine, that is, the effectiveness of RNA was revealed apparently under presence of thiamine, but was not shown in the sclerotial formation.

• The Korean Journal of Food And Nutrition
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• v.17 no.2
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• pp.177-185
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• 2004

The effects of liquid culture conditions and nutrient sources on the formation of protein-binding polysaccharide (PS) in Cordyceps militaris were examined. The formation amount of PS was increased in proportion to the growth rate of mycelium, in case of higher aeration or lower acidity. The optimum growth temperature of the mycelia was 25$^{\circ}C$ for the formation of PS. The optimum carbon source and nitrogen source were glucose and peptone, respectively. The ratio of C/N was optimal with 3％ glucose to 0.5 ％ peptone. The sugar composition in the PS was greatly changed according to the carbon sources. The mycelium of Cordyceps militaris by liquid culture showed a higher electron donating ability than that by solid culture.

• Journal of the Korean Society of Food Culture
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• v.34 no.6
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• pp.785-792
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• 2019

This study examined the changes in the protein and mineral composition of Gryllus bimaculatus fermented with Bacillus substilis and the mycelia of Basidiomycetes. Normal Gryllus bimaculatus (S) and experimental group data obtained after an inoculation of Bacillus substilis (SC) (KACC 19623), Pleurotus eryngii (SP) and Cordyceps millitaris (SC) were compared. The crude protein content of the Gryllus bimaculatus (control) was 75.48%, but it decreased to 64.55, 54.32, and 63.53% after fermentation with SB, SP and SC, respectively (p<0.05). An analysis of the organic elements showed that the contents of the carbon and nitrogen sources were also reduced after fermentation, and the most significant decrease was observed after fermentation with SP. In SDS-PAGE, a 120 kDa and a 48 kDa protein of Gryllus bimaculatus were found. On the other hand, protein bands faded after fermentation with SP and SC, respectively. Moreover, no visible band was observed after fermentation with SB. According to amino acid analysis, the total free amino acid content increased 3.84 and 1.74 times after fermentation with SB and SP, respectively, compared to the corresponding baseline data. In contrast, it decreased by 0.52 times after fermentation with SC. Among the essential amino acids found in crickets fermented with SB, the valine and isoleucine content was 3.57 and 2.64 times higher, respectively, than the recommended daily amount of essential amino acids.

• 한국균학회소식:학술대회논문집
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• 2014.10a
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• pp.19-19
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• 2014

Pleurotus ostreatus, P. eryngii, and Flammulina velutipes are major edible mushrooms that account for over 89% of total mushroom production in Korea. Recently, Agrocybe cylindracea, Hypsizygus marmoreus, and Hericium erinaceu are increasingly being cultivated in mushroom farms. In Korea, the production of edible mushrooms was estimated to be 614,224 ton in 2013. Generally, about 5 kg of mushroom substrate is needed to produce 1 kg of mushroom, and consequently about 25 million tons of spent mushroom substrate (SMS) is produced each year in Korea. Because this massive amount of SMC is unsuitable for reuse in mushroom production, it is either used as garden fertilizer or deposited in landfills, which pollutes the environment. It is reasonably assumed that SMS includes different secondary metabolites and extracellular enzymes produced from mycelia on substrate. Three major groups of enzymes such as cellulases, xylanases, and lignin degrading enzymes are involved in breaking down mushroom substrates. Cellulase and xylanase have been used as the industrial enzymes involving the saccharification of biomass to produce biofuel. In addition, lignin degrading enzymes such as laccases have been used to decolorize the industrial synthetic dyes and remove environmental pollutions such as phenolic compounds. Basidiomycetes produce a large number of biologically active compounds that show antibacterial, antifungal, antiviral, cytotoxic or hallucinogenic activities. However, most previous researches have focused on therapeutics and less on the control of plant diseases. SMS can be considered as an easily available source of active compounds to protect plants from fungal and bacterial infections, helping alleviate the waste disposal problem in the mushroom industry and creating an environmentally friendly method to reduce plant pathogens. We describe extraction of lignocellulytic enzymes and antimicrobial substance from SMSs of different edible mushrooms and their potential applications.

• Korean Journal of Microbiology
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• v.36 no.2
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• pp.97-102
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• 2000

In order to use Metarhizizmn~ anisopliae as a biological pesticide, effect of envrionmental factors on nlycelial growth, spore formation, and extracellular enzyme activity in culture broth of M. anisopliae DGUM 35001 was determined. Optimal temperature was 26^{\circ}C.$ and optimal pH ranged from 5 to 9. Among the complex media tested, MCM and SDPY media were the most favorable for mycelial growth. When Czapek-Dox agar was used as a mnimal medium, glucose and sucrose among the saccharides were very excellent source of carbohydrate. Among the biopolyners tested. chitin was the most favorable source for mycelial growth and produced high aerial inycelia. Urea and ammonium phosphate as an inorganic nitrogen source and bacto-peptone and soytone as an organic nitrogen source enhanced the mycelial growth When serine as a source of amino acid was supplemented, excellent mycelial growth was shown. Large amount of spores could be obtained from the aerial mycelia of starch medium. When the culture broth was filtrated and then the concentrate with ammonium sulfate was used as a crnde enzyme solution, high enzyme activities of amylase and protease were shown. However, lipase and chitinase activities were comparatively low.

• Korean journal of applied entomology
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• v.17 no.1 s.34
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• pp.49-51
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• 1978

The 'take-all' disease of wheat was first reported in Korea in 1947. Ophiobolnis graunis was isolated again from wheat and barley in 1976 and 1977. In 1977, perithecia of Ophiobolus were found on stems of rice, wheat and corn. Perithecia were formed on recommend varieties of wheat in the greenhouse as a result of artificial inoculation of mycelia into the soil. During 1977, Take-all of wheat and barley was quite severe. The result of survey made during this showed about 5 percent that the disease was present in Suweon, Daejeon, Yesan and Yongin. The Take-all infection causes remarkable decrease in the average weight of the grain, followed by a reduction in the number of tillers per plant and amount of gram per ear.

• Food Science and Preservation
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• v.13 no.3
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• pp.397-403
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• 2006

This experiment was conducted to examine the changes in polysaccharide concentration and morphological variation of Fomitopsis pinicola mycelium during submerged-culture in the citrus peel medium (CP). On the 12 days culture, the yields of mycelium and alcohol insoluble substance were 40.21%(w/v) and 6.94%(w/w), respectively, which were much higher than 11.29%(w/v, wet basis) and 3.17%(w/w, wet basis) obtained from YM medium. A large amount of acid soluble polysaccharides was derived from YM medium while a larger amount of alkali soluble polysaccharide was produced from CP medium. Yields of the mycelium were higher when cultured in CP medium However, there was no significant difference in formation of membranous vesicle between mycelia cultured in CP medium and YM medium. It was also observed that the formation of vacuole was closely related to the activation of the multivescular body known as cytolysome. As a result activation of mycelium and cell wall biosynthesis were more accelerated in CP medium.

• Korean journal of applied entomology
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• v.13 no.3 s.20
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• pp.105-133
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• 1974

The present study is an attempt to solve the basic problems involved in the control of the Sclerotium disease. The biologic stranis of Sclerotium rolfsii Sacc., pathogen of Sclerotium disease of Magnolia kobus, were differentiated, and the effects of vitamins, various nitrogen and carbon sources on its mycelial growth and sclerotial production have been investigated. In addition the relationship between the cultural filtrate of Penicillium sp. and the growth of Sclerotium rolfsii, the tolerance of its mycelia or sclerotia to moist heat or drought and to Benlate (methyl-(butylcarbamoy 1)-2-benzimidazole carbamate), Tachigaren (3-hydroxy-5-methylisoxazole) and other chemicals were also clarified. The results are summarizee as follows: 1. There were two biologic strains, Type-l and Type-2 among isolates. They differed from each other in the mode of growth and colonial appearance on the media, aversion phenomenon and in their pathogenicity. These two types had similar pathogenicity to the Magnolia kobus and Robinia pseudoacasia, but behaved somewhat differently to the soybaen and cucumber, the Type-l being more virulent. 2. Except potassium nitrite, sodium nitrite and glycine, all of the 12 nitrogen sources tested were utilized for the mycelial growth and sclerotial production of this fungus when 10r/l of thiamine hydrochloride was added in the culture solution. Considering the forms of nitrogen, ammonium nitrogen was more available than nitrate nitrogen for the growth of mycelia, but nitrate nitrogen was better for sclerotia formation. Organic nitrogen showed different availabilities according to compounds used. While nitrite nitrogen was unavailable for both mycelial growth and sclerotial formation whether thiamine hydrochlioride was added or not. 3. Seven kinds of carbon sources examined were not effective in general, as long as thiamine hydrochloride was not added. When thiamine hydrochloride was added, glucose and saccharose exhibited mycelial growth, while rnaltose and soluble starch gave lesser, and xylose, lactose, and glycine showed no effect at all,. In the sclerotial production, all the tested carbon sources, except lactose, were effective, and glucose, maltose, saccharose, and soluble starch gave better results. 4. At the same level of nitrogen, the amount of mycelial growth increased as more carbon Sources were applied but decreased with the increase of nitrogen above 0.5g/1. The amount of sclerotial production decreased wi th the increase of carbon sources. 5. Sclerotium rolfsii was thiamine-defficient and required thiamine 20r/l for maximun growth of mycelia. At a higher concentration of more than 20r/l, however, mycelial growth decreased as the concentration increased, and was inhibited at l50r/l to such a degree of thiamine-free. 6. The effect of the nitrogen sources on the mycelial growth under the presence of thiamine were recognized in the decreasing order of $NH_4NO_3,\;(NH_4)_2SO_4,\;asparagine,\;KNO_3$, and their effects on the sclerotial production in the order of $KNO_3,\;NH_4NO_3,\;asparagine,\;(NH_4)_2SO_4$. The optimum concentration of thiamine was about 12r/l in $KNO_3$ and about 16r/l in asparagine for the growth of mycelia; about 8r/l in $KNO_3$ and $NH_4NO_3$, and 16r/l in asparagine for the production of sclerotia. 7. After the fungus started to grow, the pH value of cultural filtrate rapidly dropped to about 3.5. Hereafter, its rate slowed down as the growth amount increased and did not depreciated below pH2.2. 8. The role of thiamine in the growth of the organism was vital. If thiamine was not added, the combination of biotin, pyridoxine, and inositol did not show any effects on the growth of the organism at all. Equivalent or better mycelial growth was recognized in the combination of thiamine+pyridoxine, thiamine+inositol, thiamine+biotin+pyridoxine, and thiamine+biotin+pyridoxine+inositol, as compared with thiamine alone. In the combinations of thiamine+biotin and thiamine+biotin+inositol, mycelial growth was inhibited. Sclerotial production in dry weight increased more in these combinations than in the medium of thiamine alone. 9. The stimulating effects of the Penicillium cultural filtrate on the mycelial growth was noticed. It increased linearly with the increase of filtrate concentration up to 6-15 ml/50ml basal medium solution. 10. $NH_4NO_3$. as a nitrogen source for mycelial growth was more effective than asparasine regardless of the concentration of cultural filtrate. 11. In the series of fractionations of the cultural filtrate, mycelial growth occured in unvolatile, ether insoluble cation-adsorbed or anion-unadsorbed substance fractions among the fractions of volatile, unvolatile acids, ether soluble organic acids, ether insoluble, cation-adsorbed, cation-unadsorbed, anion-adsorbed and anion-unadsorbed. and anion-un-adsorbed substance tested. Sclerotia were produced only in cation-adsorbed fraction. 12. According to the above results, it was assumed that substances for the mycelial growth and sclerotial formation and inhibitor of sclerotial formation were include::! in cultural filtrate and they were quite different from each other. I was further assumed that the former two substances are un volatile, ether insotuble, and adsorbed to cation-exchange resin, but not adsorbed to anion, whereas the latter is unvolatile, ether insoluble, and not adsorbed to cation or anion-exchange resin. 13. Seven amino acids-aspartic acid, cystine, glysine, histidine, Iycine, tyrosine and dinitroaniline-were detected in the fractions adsorbed to cation-exchange resin by applying the paper chromatography improved with DNP-amino acids. 14. Mycelial growth or sclerotial production was not stimulated significantly by separate or combined application of glutamic acid, aspartic acid, cystine, histidine, and glysine. Tyrosine gave the stimulating effect when applied .alone and when combined with other amino acids in some cases. 15. The tolerance of sclerotia to moist heat varied according to their water content, that was, the dried sclerotia are more tolerant than wet ones. The sclerotia harvested directly from the media, both Type-1 and Type-2, lost viability within 5 minutes at $52^{\circ}C$. Sclerotia dried for 155 days at$26^{\circ}C$ had more tolerance: sclerotia of Type-l were killed in 15 mins. at $52^{\circ}C$ and in 5 mins. at $57^{\circ}C$, and sclerotia of Type-2 were killed in 10 mins. both at $52^{\circ}C$ or $57^{\circ}C$. 16. Cultural sclerotia of both strains maintained good germinability for 132 days at$26^{\circ}C$. Natural sclerotia of them stored for 283 days under air dry condition still had good germinability, even for 443 days: type-l and type-2 maintained $20\%$ and $26.9\%$ germinability, respectively. 17. The tolerance to low temperature increased in the order of mycelia, felts and sclerotia. Mycelia completely lost the ability to grow within 1 week at $7-8^{\circ}C$> below zero, while mycelial felts still maintained the viability after .3 weeks at $7-20^{\circ}C$ below zero, and sclerotia were even more tolerant. 18. Sclerotia of type-l and type-2 were killed when dipped into the $0.05\%$ solution of mercury chloride for 180 mins. and 240 mins. respectively: and in the $0.1\%$ solution, Type-l for 60 mins. and Type-2 for 30 mins. In the $0.125\%$ uspulun solution, Type-l sclerotia were killed in 180 mins., and those of Type-2 were killed for 90 mins. in the$0.125\%$solution. Dipping into the $5\%$ copper sulphate solution or $0.2\%$ solution of Ceresan lime or Mercron for 240 mins. failed to kill sclerotia of either Type-l or Type-2. 19. Inhibitory effect on mycelial growth of Benlate or Tachi-garen in the liquid culture increased as the concentration increased. 6 days after application, obvious inhibitory effects were found in all treatments except Benlate 0.5ppm; but after 12 days, distingushed diflerences were shown among the different concentrations. As compared with the control, mycelial growth was inhibited by $66\%$ at 0.5ppm and by $92\%$ at 2.0ppm of Benlate, and by$54\%$ at 1ppm and about $77\%$ at 1.5ppm or 2.0ppm of Tachigaren. The mycelial growth was inhibited completely at 500ppm of both fungicides, and the formation of sclerotia was checked at 1,000ppm of Benlate ant at 500ppm or 1,000ppm of Tachigaren. 20. Consumptions of glucose or ammonium nitrogen in the culture solution usually increased with the increment of mycelial growth, but when Benlate or Tachigaren were applied, consumptions of glucose or ammonium nitrogen were inhibited with the increment of concentration of the fungicides. At the low concentrations of Benlate (0.5ppm or 1ppm), however, ammonium nitrogen consumption was higher than that of the ontrol. 21. The amount of mycelia produced by consuming 1mg of glucose or ammonium nitrogen in the culture solution was lowered markedly by Benlate or Tachigaren. Such effects were the severest on the third day after their treatment in all concentrations, and then gradually recovered with the progress of time. 22. In the sand culture, mycelial growth was not inhibited. It was indirectly estimated by the amount of $CO_2$ evolved at any concentrations, except in the Tachigaren 100mg/g sand in which mycelial growth was inhibited significantly. Sclerotial production was completely depressed in the 10mg/g sand of Benlate or Tachigaren. 23. There was no visible inhibitory effect on the germination of sclerotia when the sclerotia were dipped in the solution 0.1, 1.0, 100, 1.000ppm of Benlate or Tachigaren for 10 minutes or even 20 minutes.

• KSBB Journal
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• v.21 no.5
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• pp.384-388
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• 2006

The production stability of high-yielding mutants of Streptomyces lincolnensis immobilized on celite beads was examined in repeated batch cultures. We also explored the feasibility of immobilization of vegetative mycelial cells on pre-wetted celite beads, which is practical method for cell immobilization. Repeated transfer of immobilized cells into fresh medium every 10 days increased productivity of immobilized cells and maximum concentration of lincomycin, 1007 $({\pm}256)$ mg/L, was obtained at the end of the ninth cycle. A 1.4-fold higher productivity was obtained in immobilized-cell culture than that obtained by suspended-cell culture. When pre-wetted beads were inoculated with vegetative mycelia and cultured a slightly higher amount of immobilized cells and lincomycin was obtained more than those obtained by culture of spores immobilized on dry beads. This result indicates that immobilization of mycelial cells on pre-wetted beads was readily available. This technique is simple and no additional facilities are required for cell immobilization.