• Title/Summary/Keyword: protease activities

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A Study on Apoptotic Signaling Pathway in HL-60 Cells Induced by Radiation (급성전골수성백혈병 HL-60 세포주에서 방사선조사에 의한 세포고사기전)

  • Kim Hye Jung;Moon Sung Keun;Lee Jae Moon;Moon Sun Rock
    • Radiation Oncology Journal
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    • v.19 no.2
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    • pp.153-162
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    • 2001
  • Purpose : The mechanical insights of death of cancer cells by ionizing radiation are not of yet clearly defined. Recent evidences have demonstrated that radiation therapy may induce cell death via activation of signaling pathway for apoptosis in target cells. This study is designed whether ionizing radiation may activate the signaling cascades of apoptosis including caspase family cysteine pretenses, $Bcl_2/Bax$, cytochrome c and Fas/Fas-L in target cells. Materials and Methods : HL-60 cells were irradiated in vitro with 6 MV X-ray at dose ranges from 2 Gy to 32 Gy. The cell viability was tested by M assay and the extent of apoptosis was determined using agarose gel electrophoresis. The activities of caspase proteases were measured by proteolytic cleavages of substrates. Western blot analysis was used to monitor PARP, Caspase-3, Cytochrome-c, Bcl-2, Bax, Fas and Fas-L. Results : Ionizing radiation decreases the viability of HL-60 cells in a time and dose dependent manner. Ionizing radiation-induced death in HL-60 cells is an apoptotic death which is revealed as characteristic ladder-pattern fragmentation of genomic DNA over 16 Gy at 4 hours. ionizing radiation induces the activation of caspase-2, 3, 6, 8 and 9 of HL-60 cells in a time-dependent manner. The activation of caspase-3 pretense is also evidenced by the digestion of poly (ADP-ribose) polymerase and procaspase 3 with 16Gy ionizing irradiation. Anti-apoptotic Bcl2 expression is decreased but apoptotic Bax expression is increased with mitochondrial cytochrome c release in a time- dependent manner. In addiiton, expression of Fas and Fas-L is also increased in a time dependent manner. Conclusion : These data suggest that ionizing radiation-induced apoptosis is mediated by the activation of various signaling pathways including caspase family cysteine proteases, $Bcl_2/Bax$, Fas and Fas-L in a time and dose dependent manner.

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Characteristic Analysis and Production of Short-Ripened Korean Traditional Soy Sauce Added with Rice Bran (미강 첨가량에 따른 단기숙성 간장의 제조 및 특성 분석)

  • Jeong, Su-Ji;Shin, Mee-Jin;Jeong, Seong-Yeop;Yang, Hee-Jong;Jeong, Do-Youn
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.4
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    • pp.550-556
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    • 2014
  • Rice bran contains both excellent nutritional value and functional advantages. Its utilization is limited due to reducing texture and low storage. To satisfy various tastes, Bacillus spp. having high amylase and protease activities were selected. Using the strains, we made whole grain soybean Meju with a reduced manufacturing period by increasing the concentration of total nitrogen. We made soy sauces with mashing ratios of soy bean and rice bran at 10:0, 9:1, 7:3, and 5:5, and then compared their physiochemical properties. After 2 weeks of fermentation, the sugar content increased from 21~22% to 30~32%. However, pH and salinity showed no differences. At a ratio of 9:1, total nitrogen, amino nitrogen content, and total free amino acid contents were the highest at 1.62%, 652.52 mg%, and 8,804.03 mg/kg, respectively, compared to other mashing ratios of soy bean and rice bran. Especially, the contents of aspartic and glutamic acid, which increase delicate flavoring, were higher in our soy sauce compared to those of general traditional soy sauce and brewed soy sauce, which were 504.25 and 1,262.25 mg/kg, respectively. Serine and alanine, which are related to sweet taste, were present at 49.50 and 518.75 mg/kg, respectively, which were the highest among all mixing ratios, at a ratio of 9:1. Compared to general traditional soy sauce and brewed soy sauce, the contents of histamine and tyramine among biogenic amines decreased to 35.85 and 41.04 mg/kg, respectively. Finally, a soy bean and rice bran mixing ratio of 9:1 was determined to be the optimal mixing ratio in the sensory evaluation.

A Study Bioremediation of Tidal Flat by Microorganism in Pilot Scale Test (환경정화 미생물에 의한 갯벌의 생물학적 정화에 대한 파일럿 규모의 연구)

  • Choi, Hye Jin;Han, Young Sun;Park, Doo Hyun;Oh, Bo Young;Hur, Myung Je;Jo, Nam-Gyu;Kim, Young Hee;Kim, Jong-Guk
    • Journal of Life Science
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    • v.24 no.10
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    • pp.1110-1117
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    • 2014
  • Tidal flats are continuously contaminated by human activities. This study assessed the bioremediation efficiency of tidal flat soil using microcosm reactors and microorganisms originating from the tidal area. We screened 135 bacterial strains that produce extracellular enzymes from the tidal area located in the North port of Incheon bay. Two bacterial strains (Pseudoalteromonas sp. and IC35 Halothiobacillus neapolitanus IC_S22) were selected and used in the microcosm reactors, which were specially designed to functionally mimic the ecological conditions of the tidal flats. Pseudoalteromonas sp. IC35 was selected based on its relatively high activity of the enzymes amylase, cellulose, lipase, and protease. Halothiobacillus neapolitanus IC_S22 was selected for oxidation of sulfur. The M1 and M2 microcosm reactors were operated by continuous feeding of seawater under the same conditions, but M2 was first inoculated with Pseudoalteromonas sp. IC35 before the seawater feeding. The initial COD in both the M1 and M2 microcosm reactors was 320 mg/l. The final COD was 21 mg/l (M1) and 7 mg/l (M2). The M3 and M4 microcosm reactors were operated by continuous feeding of seawater under the same conditions, but M4 was first inoculated with H. neapolitanus IC_S22. The initial sulfate concentration in both the M3 and M4 microcosm reactors was 660 mg/l, and the maximum sulfate concentration was 1,360 mg/l (M3) and 1,600 mg/l (M4).

Characterization of Entomopathogenic Fungus from Trialeurodes vaporariorum and Evaluation as Insecticide (온실가루이 병원성 곰팡이의 특성 및 살충제 개발을 위한 평가)

  • Yoon, Hwi Gun;Shin, Tae Young;Yu, Mi Ra;Lee, Won Woo;Ko, Seung Hyun;Bae, Sung Min;Choi, Jae Bang;Woo, Soo Dong
    • Korean Journal of Microbiology
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    • v.49 no.1
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    • pp.64-70
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    • 2013
  • The greenhouse whitefly, Trialeurodes vaporariorum, is an economically important pest for greenhouse crops because they cause direct damage by feeding on plant nutrients and indirect damage as transmits many virus vectors. It has recently become a serious problem because of the continuous use of insecticide resulting in resistance among greenhouse whitefly population. To overcome these problems, in this study, the biological characteristics and virulence of an entomopathogenic fungus isolated from the cadaver of nymph greenhouse whitefly were investigated. Isolated fungus was identified as Isaria fumosorosea by morphological examinations and genetic identification using sequences of the ITS, ${\beta}$-tubulin, and EF1-${\alpha}$ regions. This fungus was named as I. fumosorosea SDTv and tested for the virulence against nymphs T. vaporariorum and the cold activity, the thermotolerance and the stability of UV-B irradiation on conidia. Mortality rate of greenhouse whitefly showed from 84 to 100% and the virulence increased with increasing conidial concentrations, $1{\times}10^5$ to $10^8$ conidia/ml. Conidia were stable at $35^{\circ}C$, 0.1 $J/cm^2$ of UV irradiation and germinated after 8 days at $4^{\circ}C$. Additionally, the activities of chitinases and proteases produced by I. fumosorosea SDTv were varied according to the medium. In conclusion, I. fumosorosea SDTv which showed high mortality rate against greenhouse whitefly will be used effectively in the integrated pest management programs against the greenhouse whitefly.

EFFECT OF CHLORHEXIDINE ON MICROTENSILE BOND STRENGTH OF DENTIN BONDING SYSTEMS (Chlorhexidine 처리가 상아질 접착제의 미세인장결합강도에 미치는 영향)

  • Oh, Eun-Hwa;Choi, Kyoung-Kyu;Kim, Jong-Ryul;Park, Sang-Jin
    • Restorative Dentistry and Endodontics
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    • v.33 no.2
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    • pp.148-161
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    • 2008
  • The purpose of this study was to evaluate the effect of chlorhexidine (CHX) on microtensile bond strength (${\mu}TBS$) of dentin bonding systems. Dentin collagenolytic and gelatinolytic activities can be suppressed by protease inhibitors, indicating that MMPs (Matrix metalloproteinases) inhibition could be beneficial in the preservation of hybrid layers. Chlorhexidine (CHX) is known as an inhibitor of MMPs activity in vitro. The experiment was proceeded as follows: At first, flat occlusal surfaces were prepared on mid-coronal dentin of extracted third molars. GI (Glass Ionomer) group was treated with dentin conditioner, and then, applied with 2 % CHX. Both SM (Scotchbond Multipurpose) and SB (Single Bond) group were applied with CHX after acid-etched with 37% phosphoric acid. TS (Clearfil Tri-S) group was applied with CHX, and then, with adhesives. Hybrid composite Z-250 and resin-modified glass ionomer Fuji-II LC was built up on experimental dentin surfaces. Half of them were subjected to 10,000 thermocycle, while the others were tested immediately. With the resulting data, statistically two-way ANOVA was performed to assess the ${\mu}TBS$ before and after thermo cycling and the effect of CHX. All statistical tests were carried out at the 95 % level of confidence. The failure mode of the testing samples was observed under a scanning electron microscopy (SEM). Within limited results, the results of this study were as follows; 1. In all experimental groups applied with 2 % chlorhexidine, the microtensile bond strength increased, and thermo cycling decreased the micro tensile bond strength (P > 0.05). 2. Compared to the thermocycling groups without chlorhexidine, those with both thermocycling and chlorhexidine showed higher microtensile bond strength, and there was significant difference especially in GI and TS groups. 3. SEM analysis of failure mode distribution revealed the adhesive failure at hybrid layer in most of the specimen. and the shift of the failure site from bottom to top of the hybrid layer with chlorhexidine groups. 2 % chlorhexidine application after acid-etching proved to preserve the durability of the hybrid layer and microtensile bond strength of dentin bonding systems.

Fungal Microflora on Korean Home-made Meju (재래식(在來式) 메주에 분포(分布)하고 있는 진균(眞菌)에 관한 조사(調査) 연구(硏究))

  • Park, Kyoung-Ja;Kim, Young-Mi;Lee, Bae-Ham;Lee, Bok-Kwon
    • The Korean Journal of Mycology
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    • v.5 no.1
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    • pp.7-12
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    • 1977
  • For this study 3 samples of the home-made meju and 3 samples of the improved meju were selected according to their characteristics. A total of 25 strains of true fungi were isolated from the samples of the home-made meju and identified by the Key of Alexopoulos and Raper, using a strain Aspergillus oryzae A-113 as a control. Amylolytic and proteolytic enzyme activities of the isolated strains were investigated ana the results obtained were as follows. 1. The 25 strains from the home-made meju were identified into 2 Aspergillus oryzae, 14 Asp. flavus, 6 Penicillum spp. 1 Candida sp 1 Spicaria sp and 1 Rhizopus sp. 2. The 3 strains from the improved meju were all identified as Aspergillus oryzae. 3. Aspergillus flavus, A-B, from the home-made meju was found to he the strongest strain in ${\alpha}-amylase$ activity and also to be similar to the strains of Aspergillus orzae from improved meju. 4. Aspergillus flavus, A-7, from the home-made meju was found to be the strogest strain in ${\beta}-amylase$ activity and stronger than that from the improved meju. 5. Aspergillus flavus B-3, was found to be the strongest strain in protease activity and stronger than that from the improved meju. 6. Some of the strains from the home-made meju turned out to be harmful strains, such as Penicillium spp. which secrete antibiotics, Asp. flavus which secretes mycotoxin, Candida sp which causes skin diseases, Spicaria sp. which is a insect pathogen. 7. Rhizopus sp was also found but it has not been proved to be harmful.

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Studies on the Mutation of Aspergillus niger (흑국균(黑麴菌)의 인공변이(人工變異)에 관(關)한 연구(硏究))

  • Park, Yoon-Joong;Sohn, Cheon-Bae
    • Korean Journal of Food Science and Technology
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    • v.14 no.1
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    • pp.72-79
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    • 1982
  • Several mutants were isolated from the parent strain of Aspergillus niger CF: the first mutant strain CF-11 was obtained by UV irradiation, and the second mutant strain CF-21 and CF-22 were from NTG (N-methyl-N'-nitroso-N-nitroso-guanidine) treatment on the CF-11. These mutants were characterized, and their enzyme and acid production on wheat bran Koji and wheat flour Koji were studied. Asp. niger CF-22 mutant appeared to be tan type which conidial heads were discolored. It's glucoamylase activity was inreased approximately two times and its ${\alpha}-amylase$ about 50 percent as compared with that of the parent strain of Asp. niger CF, when grown on wheat bran Koji under the optimal conditions. Asp. niger CF-21 mutant showed slower growth rate and poor sporulation than the wild type, although its conidial heads were not discolored. Approximately 4-fold increment in its acid production was observed as compared with the weld type. The activities of glucoamylase and ${\alpha}-amylase$ of the Asp. niger CF-22 and CF-21 mutants were higher than those of the wild type, but their protease activity was rather lower. The maximum production of glucoamylase by the Asp. niger CF-22 mutant was obtained after 2 to 3 days incubation on wheat bran at 30 to $35^{\circ}C;$ ${\alpha}-amylase$2 days incubation at 30 to $35^{\circ}C$. The maximal levels of acid production by the mutant CF-21 was appeared after 2 days incubation on wheat bran Koji, and after 3 days on wheat flour Koji at $30^{\circ}C$. Little differences in the levels of acid production were observed between on wheat bran and flour Koji.

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Characteristics of Ju-Back and Effect of Ju-Back Fertilizer on Growth of Crop Plants (주류생산 부산물인 주박의 특성 규명 및 주박이 작물생육에 미치는 영향)

  • Lee, Jung-Hoon;Park, Sung-Min;Park, Chi-Duck;Jung, Hyuck-Jun;Kim, Hyun-Soo;Yu, Tae-Shick
    • Journal of Life Science
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    • v.17 no.11
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    • pp.1562-1570
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    • 2007
  • This experiment was conducted to develop fertilizer which promotes plant growth as well as suppressing pathogenic fungi. The fertilizer was made from the mixture of Ju-Back (Korean rice wine cake) and indigenous rhizosphere-bacterium. The main ingredients of Ju-Back were investigated as 6.04% total nitrogen, 42.59% total carbohydrate, 1.01% available phosphate, 73.42% organic matter, 7.72% potassium oxide, 1.35% calcium oxide, 0.53% magnesium oxide. The enzyme activities of Ju-Back were estimated to be 980 units/g for ${\alpha}-amylase$, 300 units/g for glucoamylase, and 1800 units/g for acid pretense. Indigenous rhizosphere bacteria which produced antifungal agent were isolated from soil, and was selected KMU-13 strain which can antagonize against various plant pathogenic fungi (Botrytis cinerea KACC 40573, Sclerotinia sclerotiorum KACC 41065, Fusairum oxysporum KACC 40052, Pythium aphanidermatum KACC 40156, Phytophthora capsici KACC 40476 and Glomerella cingulata KACC 40299). KMU-13 strain was identified as Bacillus subtilis KMU-13 by biochemical and 16s rDNA analysis. The organic fertilizer was made as prototype which was composed 20% Ju-Back, 70% carrier, 9.7% microorganism cultivated solution, 0.3% trace-element. We also investigated an application of fertilizer using Ju-Back for cultivating lettuce (Lactuca sativar) which were grown in three soil conditions that had chemical fertilizer, barnyard manure, lime power, urea, potassium chloride and superphosphate as a control, the whole quantity (80 kg/10a) of posted fertilizer with the control and the half quantity (40 kg/10a) with the control. The growth characteristics were examined and analysed with several weeks interval from 3 weeks to 8 weeks on head length (cm), head width (cm/head), number of leaf and fresh weight (g/plant). The results are summarized as follows. The head width and fresh weight of lettuce were the highest at posted fertilizer 1 (whole quantity) was applied chemical, organic matter (Ju-Back) and carrier. The head length was the highest at posted fertilizer 2 (whole quantity) was applied Ju-Back only.

Comparison of Antibacterial and Antioxidant Activities of Mushroom Mycelium Culture Extracts Cultivated in the Citrus Extracts (감귤농축액 첨가배지에서 배양한 버섯균사체 추출물의 항균활성 및 항산화활성 비교)

  • Kim Man-Chul;Kim Min-Joo;Kim Taeg;Park Guen-Tae;Son Hong-Joo;Kim Gi-Young;Choi Woo-Bong;Oh Duck-Chul;Heo Moon-Soo
    • KSBB Journal
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    • v.21 no.1 s.96
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    • pp.72-78
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    • 2006
  • This study was carried out to investigate the antimicrobial and antioxidative effects of mycelium cultural extract from mushroom. Mushroom mycelium was grown in a defined synthetic liquid medium and citrus extracts, and the culture extracts were examined for antioxidant and antibacterial activity. Myceliums of Phellinus linteus, Cordyceps militaris, Coriolus versicolor, Sparassic crispa, Agaricus blazei, lnonotus obliquus, Lentinus edodes, Hericium erinacium, Gonoderma lucidium in 10% citrus extract supplemented medium and synthesis medium were incubated in a shaking incubator (120 rpm, $24{\sim}30^{\circ}C$ ) for $7{\sim}15$ days. The antimicrobial activity of the culture fluid of mushroom mycelium grown in submerged liquid culture was tested against 12 microorganisms which were fish pathogens and common bacterial species. The culture extracts showed high activity against Vibrio sp. and had poor effect on Streptocouus sp., S. parauberis, S. iniae. The culture extracts obtained from the synthetic medium showed $30{\sim}93%$ of the 1,1-diphenyl-2-picrylhydrazyl radical scavenger activity, the culture extracts obtained from the citrus extracts medium exhibited antioxidant activity up to 55%.

Studies on the physio-chemical properties and the cultivation of oyster mushroom(Pleurotus ostreatus) (느타리버섯의 생리화학적성질(生理化學的性質) 및 재배(栽培)에 관(關)한 연구(硏究))

  • Hong, Jai-Sik
    • Applied Biological Chemistry
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    • v.21 no.3
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    • pp.150-184
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    • 1978
  • Nutritional characteristics and physio-chemical properties of mycelial growth and fruitbody formation of oyster mushroom(Pleurotus ostreatus)in synthetic media, the curtural condition for the commerical production in the rice straw and poplar sawdust media, and the changes of the chemical components of the media and mushroom during the cultivation were investigated. The results can be summarized as follows: 1. Among the carbon sources mannitol and sucrose gave rapid mycelial growth and rapid formation of fruit-body with higher yield, while lactose and rhamnose gave no mycelial growth. Also, citric acid, succinic acid, ethyl alcohol and glycerol gave poor fruit-body formation, and acetic acid, formic acid, fumaric acid, n-butyl alcohol, n-propyl alcohol and iso-butyl alcohol inhibited mycelial growth. 2. Among the nitrogen sources peptone gave rapid mycelial growth and rapid formation of fruit-body with higher yield, while D,L-alanine, asparatic acid, glycine and serine gave very poor fruit-body formation, and nitrite nitrogens, L-tryptophan and L-tyrosine inhibited mycelial growth. Inorganic nitrogens and amino acids added to peptone were effective for fruit-body growth, and thus addition of ammonium sulfate, ammonium tartarate, D,L-alanine and L-leucine resulted in about 10% increase fruit-body yield. L-asparic acid about 15%, L-arginine about 20%, L-glutamic acid, and L-lysine about 25%. 3. At C/N ratio of 15.23 fruit-body formation was fast, but the yield decreased, and at C/N ratio of 11.42 fruit-body formation was slow, but the yield increased. Also, at the same C/N ratio the higher the concentration of mannitol and petone, the higher yield was produced. Thus, from the view point of both yield of fruit-body and time required for fruiting the optimum C/N ratio would be 30. 46. 4. Thiamine, potassium dihydrogen phosphate and magnecium sulfate at the concentration of $50{\mu}g%$. 0.2% and 0.02-0.03%, respectively, gave excellent mycelial and fruit-body growth. Among the micronutrients ferrous sulfate, zinc sulfate and manganese sulfate showed synergetic growth promoting effect but lack of manganese resulted in a little reduction in mycelial and fruit-body growth. The optimum concentrati on of each these nutrients was 0.02mg%. 5. Cytosine and indole acetic acid at 0.2-1mg% and 0.01mg%, respectively, increased amount of mycelia, but had no effect on yield of fruit-body. The other purine and pyrimidine bases and plant hormones also had no effect on mycelial and fruit-belly yield. 6. Illumination inhibited mycelial growth, but illumination during the latter part of vegetative growth induced primordia formation. The optimum light intensity and exposure time was 100 to 500 lux and 6-12 hours per day, respectively. Higher intensity of light was injurous, and in darkness only vegetative growth without primordia formation was continued. 7. The optimum temperature for mycelial growth was $25^{\circ}C$ and for fruit-body formation 10 to $15^{\circi}C$. The optimum pH range was from 5.0 to 6.5. The most excellent fry it-body formation were produced from the mycelium grown for 7 to 10 days. The lesser the volume of media, the more rapid the formation of fruit-body; and the lower the yield of fruit-body; and the more the volume of media, the slower the formation of fruit-body, and the higher the yield of fruit-body. The primordia formation was inhibited by $CO_2$. 8. The optimum moisture content for mycelial growth was over 70% in the bottle media of rice straw and poplar sawdust. 10% addition of rice bran to the media exhibited excellent mycelial growth and fruit-body formation, and the addition of calciumcarbonate alone was effective, but the addition of calcium carbonate was ineffective in the presence of rice bran. 9. In the cultivation experiments the total yield of mushroom from the rice straw media was $14.99kg/m^2$, and from the sawdust media $6.52kg/m^2$, 90% of which was produced from the first and second cropping period. The total yield from the rice straw media was about 2.3 times as high as that from the sawdust media. 10. Among the chemical components of the media little change was observed in the content of ash on the dry weight basis, and organic matter content decreased as the cultivation progressed. Moisture content, which was about 79% at the time of spawning, decreased a little during the period of mycelial propagation, after which no change was observed. 11. During the period from spawning to the fourth cropping about 16.7% of the dry matter, about 19.3% of organic matter, and about 40% of nitrogen were lost from the rice straw media; about 7.5% of dry mallet, about 7.6% of organic matter, and about 20% of nitrogen were lost from the sawdust media. For the production of 1kg of mushroom about 232g of organic matter and about 7.0g of nitrogen were consumed from the rice straw media; about 235g of organic matter and about 6.8g of nitrogen were consumed from the sawdust media, 1㎏ of mushroom from either of media contains 82.4 and 82.3g of organic matter and 5.6 and 5.4g of nitrogen, respectively. 12. Total nitrogen content of the two media decreased gradually as the cultivation progressed, and total loss of insoluble nitrogen was greater than that of soluble nitrogen. Content of amino nitrogen continued to increase up to the third cropping time, after which it decreased. 13. In the rice straw media 28.0 and 13.8% of the total pentosan and ${\alpha}$-cellulose, respectively, lost during the whole cultivation period was lost during the period of mycelial growth; in the sawdust media 24.1 and 11.9% of the total pentosan and ${\alpha}$-cellulose, respectively, was lost during the period of mycelial growth. Lignin content in the media began to decrease slightly from the second cropping time, while the content of reduced sugar, trehalose and mannitol continued to increase. C/N ratio of the rice straw media decreased from 33.2 at spawining to 30.0 at ending; that of the sawdust media decreased from 61.3 to 60.0. 14. In both media phosphorus, potassium, manganese and zinc decreased, at magnesium, calcium and copper showed irregular changes, and iron had a tendency to be increased. 15. Enzyme activities are much higher in the rice straw media than in the sawdust media. CMC saccharifying and liquefying activity gradually increased from after mycelial propagation to the second cropping, after which it decreased in both media. Xylanase activity rapidly and greatly increased during the second cropping period rather than the first period. At the start of the third cropping period the activity decreased rapidly in the rice straw media, which was not observed in the sawdust media. Protease activity was highest after mycelial propagation, after which it gradually decreased. The pH of the rice straw media decreased from 6.3 at spawning to 5.0 after fourth cropping; that of the sawdust media decreased from 5.7 to 4.9. 16. The contents of all the components except crude fibre of the mushroom from the rice straw media were higher than those from the sawdust media. Little change was observed in the content of the components of mushroom cropped from the first to the third period, but slight decrease was noticed at the fourth cropping.

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