• Applied Biological Chemistry
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• v.19 no.2
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• pp.82-92
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• 1976

The study was carried out to investigate the changes of the various chemical components and the microflora during the aging period of Korean navive Kochuzang. (Red pepper soybean paste) Korean native maeju loaves were separated into surface and inner parts. Three kinds of Korean native Kochuzang were prepared from surface part, inner part, and ordinary of maeju. The selection and the indentification of the high enzyme producing strains from the microflora and characteristics of their enzymes were studied. I. The changes of the various chemical components during the aging period of Kochuzang. 1) The changes of pH in the 3 kinds of Kochuzang displayed rapid decrease for the first 10 days after preparing and gradual curve of decrease until 60 days, but slight increase for the next 30 days. The pH of the surface part Kochuzang was lower than that of inner part or ordinary Kochuzang. 2) The total acid contents in the 3 kinds of Kochuzang showed gradual increase until the 60 days but it slowly reduced after this time. 3) The total nitrogen contents in the 3 kind of Kochuzang showed gradual inerease up to the 60 days, but slight decrease after this time. 4) The changes of trichloroacetic acid soluble nitrogen in the 3 kinds of Kochuzang showed a remarkable increase for the first 10 days, however gradual increase after this time. 5) The increase of amino nitrogen contents in the 3 kinds of Kochuzang seemed to be remarkable until the first 30 days, however to be less remarkable after this time. 6) The contents of reducing sugar in the 3 kinds of Kochuzang showed remarkable increase until the first 50 days and it slowly reduced after this time. II. The changes of microflora during the aging period of Kochuzang. 1) Aerobic, anaerobic bacteria and mold in the 3 kinds of Kochuzang were increased until the first 30 to 40 days, but they were reduced after this time. 2) No yeast in the three kinds of Kochuzang appeared until the first 20 days. Yeast were proved to grow, when the pH value was decreased below 5.4 after the 30 days. Yeasts in the surface part and ordinary Kochuzang were gradually increased and those in the inner part Kochuzang were decreased as aging. III. The selection and identification of high amylase and protease producing strains from the microflora during the aging period of Kochuzang. 1) The amylase and protease highly producing strains from microflora were identified as Bacillus subtilis-P, Bacillus subtilis-G, Bacillus licheniformis-K, Aspergillus oryzae-B. 2) Amylase activity of Aspergillus oryzae-B was highest among the strains and the strains in order of the higher activity to the lower one were Bacillus subtilis-P Bacillus licheniformis-K, Bacillus subtilis-G. Protease activities of Aspergillus oryzae-B and Bacillus subtilis-P were about the same and the strains in order of the higher activity to the lower one were Bacillus licheniformis-K, Bacillus subtilis-G. 3) Amylase activity was inhibited more than protease activity was with NaCl concentration. Amylase activity was inhibited by 45 to 65 percent and protease activity by 40 to 46 percent at the concentration of 15 percent NaCl, which was the average concentration of NaCl in Kochuzang.

• Korean Journal of Veterinary Research
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• v.35 no.3
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• pp.543-552
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• 1995

This study was undertaken to investigate the eftects of Cd administered ad libitum for 6 weeks on the cellular immune responses of Balb/c mice. The results were summarized as follows; 1. The mice fed 25, 50 and 100ppm Cd drank as much as control, but the mice fed 200ppm Cd drank significantly less water after Cd exposure than did control. Increasing rates of body weight of Cd-fed mice for 6 weeks were as this, control group 27.0%, Cd administered groups(25, 50, 100 and 200ppm) 28.54%, 28.31%, 20.49% and 18.04%, respectively. 2. Absolute spleen to body weight(mg/g) of control, 25, 50, 100 and 200ppm Cd administered groups were $4.34{\pm}0.23$, $4.20{\pm}0.54$, $4.80{\pm}0.87$, $4.25{\pm}0.32$ and $4.40{\pm}0.32$, respectively. Splenic cellularity(${\times}10^7$) of control was $24.29{\pm}5.98$ but increased to $27.72{\pm}5.48$, $32.96{\pm}8.44$, $28.32{\pm}8.76$ and $29.64{\pm}4.08$ in 25, 50, 100 and 200ppm Cd-fed groups, respectively. 3. Total $CD_4{^+}$ cells(${\times}10^7$) of control, 25, 50, 100 and 200ppm Cd-fed groups were $9.15{\pm}2.24$, $10.40{\pm}2.04$, $12.04{\pm}3.08$, $10.20{\pm}3.16$ and $10.80{\pm}1.48$, respectively and total $CD_8{^+}$ cells(${\times}10^7$) of these groups were $2.32{\pm}0.56$, $2.54{\pm}0.27$, $3.12{\pm}0.80$, $2.25{\pm}0.70$ and $2.24{\pm}0.28$, in order. On the other hand, $CD_4{^+}/CD_8{^+}$ ratios in total cells were increased significantly except for 50ppm Cd-fed group($3.88{\pm}0.01$). And that of control was $3.97{\pm}0.02$, but those of 25, 100 and 200ppm were $4.35{\pm}0.01$, $4.54{\pm}0.03$ and $4.81{\pm}0.03$. 4. Phagocytosis rates of peritoneal macrophages were increased significantly in 25 and 50ppm Cd groups($36.34{\pm}9.45$ and $37.15{\pm}9.22$, respectively), but 100 and 200ppm groups showed similar rates($18.20{\pm}3.04$ and $19.48{\pm}3.22$ respectively) to that of control($21.43{\pm}3.62$). 5. In mitogen-induced splenocyte proliferation, various concentraions of $CdCl_2(10^{-4}-10^{-7}M)$ were added to mitogen-stimulated culture in vitro. Splenocyte proliferation induced by LPS was decreased dose dependently, but proliferation by Con-A was increased slightly in concentrations of $10^{-7}-10^{-6}M$. 6. Significant cytotoxicity of splenocytes with $CdCl_2$ were shown at $10^{-4}M$ treated group, especially at 24 hrs. From these results, it could be concluded that Cd might modulate the immune responses by modifying a distribution of T cell subpopulations.

• Applied Biological Chemistry
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• v.10
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• pp.69-100
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• 1968

1. In order to investigate on the microflora and enzyme activity of mold wheat 'Nuruk' , the major source of microorganisms for the brewing of Takju (a Korean Sake), two samples of Nuruk, one prepared at the College of Agriculture, Chung Nam University (S) and the other perchased at a market (T), were taken for the study. The molds, aerobic bacteria, lactic acid bacteria, and yeasts were examined and counted. The yeasts were classified by the treatment with TTC (2, 3, 5 triphenyltetrazolium chloride) agar that yields a varied shade of color. The amylase and protease activities of Nuruk were measured. The results were as the followings. a) In the Nuruk S found were: Aspergillus oryzae group, $204{\times}10^5$; Black Aspergilli, $163{\times}10^5$; Rhizogus, $20{\times}10^5$; Penicillia, $134{\times}10^5$; Areobic bacteria, $9{\times}10^6-2{\times}10^7$; Lactic acid bacteria, $3{\times}10^4$ In the Nuruk T found were: Aspergillus oryzae group, $836{\times}10^5$; Black Aspergilli, $286{\times}10^5$; Rhizopus, $623{\times}10^5$; Penicillia, $264{\times}10^5$; Aerobic bacteria, $5{\times}10^6-9{\times}10^6$; Lactic acid bacteria, $3{\times}10^4$ b) Eighty to ninety percent of the aerobic bacteria in Nuruk S appeared to belong to Bacillus subtilis while about 70% of those in Nuruk T seemed to be spherical bacteria. In both Nuruks about 80% of lactic acid bacteria were observed as spherical ones. c) The population of yeasts in 1g. of Nuruk S was about $6{\times}10^5$, 56.5% of which were TTC pink yeasts, 16% of which were TTC red pink yeasts, 8% of which were TTC red yeasts, 19.5% of which were TTC white yeasts. In Nuruk T(1g) the number of yeasts accounted for $14{\times}10^4$ and constituted of 42% TTC pink. 21% TTC red pink 28% TTC red and 9% TTC white. d) The enzyme activity of 1g Nuruk S was: Liquefying type Amylase, $D^{40}/_{30},=256$ W.V. Saccharifying type Amylase, 43.32 A.U. Acid protease, 181 C.F.U. Alkaline protease, 240C.F.U. The enzyme activity of 1g Nuruk T was: Liquefying type Amylase $D^{40}/_{30},=32$ W.V. Saccharifying type amylase $^{30}34.92$ A.U. Acid protease, 138 C.F.U. Alkaline protease 31 C.F.U. 2. During the fermentation of 'Takju' employing the Nuruks S and T the microflora and enzyme activity throughout the brewing were observed in 12 hour intervals. TTC pink and red yeasts considered to be the major yeasts were isolated and cultured. The strains ($1{\times}10^6/ml$) were added to the mashes S and T in which pH was adjusted to 4.2 and the change of microflora was examined during the fermentation. The results were: a) The molds disappeared from each sample plot since 2 to 3 days after mashing while the population of aerobic bacteria was found to be $10{\times}10^7-35{\times}10^7/ml$ inS plots and $8.2{\times}10^7-12{\times}10^7$ in plots. Among them the coccus propagated substantially until some 30 hours elasped in the S and T plots treated with lactic acid but decreased abruptly thereafter. In the plots of SP. SR. TP. and TR the coccus had not appeared from the beginning while the bacillus showed up and down changes in number and diminished by 1/5-1/10 the original at the end stage. b) The lactic acid bacteria observed in the S plot were about $7.4{\times}10^7$ in number per ml of the mash in 24 hours and increased up to around $2{\times}10^8$ until 3-4 days since. After this period the population decreased rapidly and reached about $4{\times}10^5$ at the end, In the plot T the lactic acid becteria found were about $3{\times}10^8$ at the period of 24 fours, about $3{\times}10$ in 3 days and about $2{\times}10^5$ at the end in number. In the plots SP. SR. TP, and TR the lactic acid bacteria observed were as less as $4{\times}10^5$ at the stage of 24 hours and after this period the organisms either remained unchanged in population or ceased to exist. c) The maiority of lactic acid bacteria found in each mash were spherical and the change in number displayed a tendency in accordance with the amount of lactic acid and alcohol produced in the mash. d) The yeasts had showed a marked propagation since the period of 24 hours when the number was about $2{\times}10^8$ ㎖ mash in the plot S. $4{\times}10^8$ in 48 hours and $5-7{\times}10^8$ in the end period were observed. In the plot T the number was $4{\times}10^8$ in 24 hours and thereafter changed up and down maintaining $2-5{\times}10^8$ in the range. e) Over 90% of the yeasts found in the mashes of S and T plots were TTC pink type while both TTC red pink and TTC red types held range of $2{\times}10-3{\times}10^7$ throughout the entire fermentation. f) The population of TTC pink yeasts in the plot SP was as $5{\times}10^8$ much as that is, twice of that of S plot at the period of 24 hours. The predominance in number continued until the middle and later stages but the order of number became about the same at the end. g) Total number of the yeasts observed in the plot SR showed little difference from that of the plot SP. The TTC red yeasts added appeared considerably in the early stage but days after the change in number was about the same as that of the plot S. In the plot TR the population of TTC red yeasts was predominant over the T plot in the early stage which there was no difference between two plots there after. For this reason even in the plot w hers TTC red yeasts were added TTC pink yeasts were predominant. TTC red yeasts observed in the present experiment showed continuing growth until the later stage but the rate was low. h) In the plot TP TTC pink yeasts were found to be about $5{\times}10^8$ in number at the period of 2 days and inclined to decrease thereafter. Compared with the plot T the number of TTC pink yeasts in the plot TP was predominant until the middle stage but became at the later stage. i) The productivity of alcohol in the mash was measured. The plot where TTC pink yeasts were added showed somewhat better yield in the earely stage but at and after the middle stage the difference between the yeast-added and the intact mashes was not recognizable. And the production of alcohol was not proportional to the total number of yeasts present. j) Activity of the liquefying amylase was the highest until 12 hours after mashing, somewhat lowered once after that, and again increased around 36-48 hours after mashing. Then the activity had decreased continuously. Activity of saccharifying amylase also decreased at the period of 24 hours and then increased until 48 hours when it reached the maximum. Since, the activity had gradually decreased until 72 hours and rapidly so did thereafter. k) Activity of alkaline protease during the fermentation of mash showed a tendency to decrease continusously although somewhat irregular. Activity of acid protease increased until hours at the maximum, then decreased rapidly, and again increased, the vigor of acid protease showed better shape than that of alkaline protease throughout. 3. TTC pink yeasts that were predominant in number, two strains of TTC red pink yeasts that appeared throughout the brewing, and TTC red yeasts were identified and the physiological characters examined. The results were as described below. a) TTC pinkyeasts (B-50P) and two strains of TTC red pink yeasts (B-54 RP & B-60 RP) w ere identified as the type of Saccharomyces cerevisiae and TTC pink red yeasts CB-53 R) were as the type of Hansenula subpelliculosa. b) The fermentability of four strains above mentioned were measured as follows. Two strains of TTC red pink yeasts were the highest, TTC pink yeasts were the lowest in the fermantability. The former three strains were active in the early stage of fermentation and found to be suitable for manufacturing 'Takju' TTC red yeasts were found to play an important role in Takju brewing due to its strong ability to produce esters although its fermentability was low. c) The tolerance against nitrous acid of strains of yeast was marked. That against lactic acid was only 3% in Koji extract, and TTC red yeasts showed somewhat stronger resistance. The tolerance against alcohol of TTC pink and red pink yeasts in the Hayduck solution was 7% while that in the malt extract was 13%. However, that of TTC red yeasts was much weaker than others. Liguefying activity of gelatin by those four strains of yeast was not recognized even in 40 days. 4. Fermentability during Takju brewing was shown in the first two days as much as 70-80% of total fermentation and around 90% of fermentation proceeded in 3-4 days. The main fermentation appeared to be completed during :his period. Productivity of alcohol during Takju brewing was found to be apporximately 65% of the total amount of starch put in mashing. 5. The reason that Saccharomyces coreanuss found be Saito in the mash of Takju was not detected in the present experiment is considered due to the facts that Aspergillus oryzae has been inoculated in the mold wheat (Nuruk) since around 1930 and also that Koji has been used in Takju brewing, consequently causing they complete change in microflora in the Takju brewing. This consideration will be supported by the fact that the original flavor and taste have now been remarkably changed.

• Applied Biological Chemistry
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• v.17 no.2
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• pp.81-92
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• 1974

In order to prepare the mashing materials for 'Takju', Korean wine, with potatoes, theywere steamed, dryed, and pulverized, and their chemical components were analyzed. As a brewing method of Takju with potatoes, general 2nd stage process with Ipkuk and Bunkuk (enzyme sources), commonly used now, was carried out and the effects of preparing conditions of Ipkuk(koji) with potato flour, mashing materials and brewing conditions on the contents of Takju mash, and of storing time on the contents of Takju, were investigated and the results obtained were summarized as follows, 1. Chemical components of steamed potatoes and potato flour were Moisture; 76.2, 10.8%, Total sugar; 16.1, 69.8%, Reducing sugar; 3.45, 13.4%, Crude protein; 2.1, 11.3%, Total acid; 0.012, 0.023% and Volatile acid; 0.0012, 0.0025% respectively. 2. The most effective preparing conditions of Ipkuk with potato flour were to incubate the potato flour added 40-50% of water for 48 hours by general preparing process of Koji, and liquefying and saccharogenic amylase activities of Ipkuk incubated at above conditions were $D40^{\circ}$ 30' 128 W.V. and 13.2 A.U.. 3. The effects of various brewing conditions on the contents of Takju mashes were as follows; 1) Optimum ratio of mashing water and materials for Takju brewing with potato flour was 140ml of water to 60g of flour in 1st stage and 260ml to 140g in 2nd stage. 2) Optimum fermentating times and temperatures for Takju brewing were at $25^{\circ}C$ for 48 hours in 1st stage and at $30^{\circ}C$ for 48 hours in 2nd stage. 3) Optimum amounts of enzyme sources for Takju brewing were 20-30% of Ipkuk and 0.5% of Bunkuk in 1st stage and 1.0% of Bunkuk in 2nd stage. 4) Methanol content of the Takju mash brewed with raw potato flour was much more than that with steamed potato flour. 5) Alcohol, fusel oil and Formol nitrogen contents of the Takju mash brewed with potato flour were less than that with wheat flour, on the contrary, methanol contents and total acidifies of them were showed conversely above. 4. The changes of chemical components and microflora in the mashes during the brewing potato flour Takju were as follows; 1) The accumulation of ethanol followed rapidly in early stage, being the highest at 72 hours (11.9%). 2) Total sugar content of the mash was decreased considerably within 48-72 hours, being 2.62% at 72 hours, and thereafter slowly. 3) Reducing sugar of the mash had a tendency of decreasing, being 0.29% at 48 hours. 4) Total acidity, volatile acidity and Formol nitrogen content of the mash were increased slowly, being 7.30, 0.20, 2.55 at 48 hours. 5) Total cells of yeast appeared the highest in 72 hours ($2.1{\times}10^8$) and thereafter decreased slowly. 6) Total cells of bacteria appeared the highest in 48 hours ($2.4{\times}10^8$) and thereafter decreased or increased slightly. 5. Takju was made from the fermented mash mixed with water to be 6% of alcohol content, and the change of alcohol content, total acidity, total cells of yeast and bacteria during the storing at $30^{\circ}C$ were as follows; 1) Alcohol content of Takju was increased slightly at 24 hours (6.2%), and thereafter decreased slowly. 2) Total acidity of Takju was increased gradually, being 6.1 at 72 hours 3) Total cells of yeast and bacteria appeared the highest at 48 hours ($2.3{\times}10^8,\;1.5{\times}10^8$), and thereafter decreased slowly. 6. Alcohol content, total acidity and Formol nitrogen content of the Takju brewed with potato flour Ipkuk or wheat flour Ipkuk and steamed potatoes(1:5) were 9.8-11.3%, 5.8-7.4, 2.5-3.3 respectively, and the color of the Takju. was similar to commercial Takju. 7. The results of sensory test for various experimental Takju, showed that the Takjues brewed with the materials combined with wheat flour and steamed potatoes(4:5 or 3.5:7.5) were not significantly different in color, taste and flavor from commercial Takju, However, those with potato flour and wheat flour (1:1 or 7:3) were significantly different from commercial Takju.

• Korean Journal of Agricultural Science
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• v.1 no.1
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• pp.67-81
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• 1974

In order to prepare the mashing materials for "Takju", Korean wine, with potatoes they were steamed, dryed, and pulverized, and their chemical components were analyzed. As a brewing method of Takju with potatoes, general 2nd stage process with Ipkuk and Bunkuk (enzyme sources), commonly used now, was carried out and the effects of preparing conditions of Ipkuk(koji) with potato flour, mashing materials and brewing conditions on the contents of Takju mash and of storing time on the contents of Takju, were investigated and the results obtained were summarized as follows. 1. Chemical components of steamed potatoes and potato flour were Moisture; 76.2, 10.8%, Total sugar; 16.1, 69.8%, Reducing sugar; 3.45, 13.4%, Crude protein; 2.1, 11.3%, Total acid; 0.012, 0.023% and Volatile acid; 0.0012, 0.0025% respectively 2. The most effective preparing conditions of Ipkuk with potato flour were to incubate the potato flour added 40-50% of water for 48 hours by general preparing process of Koji, and liquefying and saccharogenic amylase activities of Ipkuk incubated at above conditions were $D_{40^{\circ}}{^{30{\prime}}}$ 128 W.V. and 13.2 A. U. 3. The effects of various brewing conditions on the contents of Takju mashes wereas follows; 1) Optimum ratio of mashing water and materials for Takju brewing with potato flour was 140ml of water to 60g of flour in 1st stage and 260ml to 140g in 2nd stage. 2) Optimum fermentating times and temperatures for Takju brewing were at $25^{\circ}C$ for 48 hours in 1st stage and at $30^{\circ}C$ for 48 hours in 2nd stage. 3) Optimum amounts of enzyme sources for Takju brewing 20-30% of Ipkuk and 0.5% of Bunkuk in 1st stage and 1.0% of Bunkuk in 2nd stage. 4) Methanol content of the Takju mash brewed with raw potato flour was much more than that with steamed potato flour. 5) Alcohol fusel oil and Formal nitrogen contents of the Takju mash brewed with potato flour were less than that with wheat flour, on the contrary, methanol contents and total acidities of them were showed conversely above. 4. The changes of chemical components and microflora in the mashes during the brewing potato flour Takju were as follows; 1) The accumulation of ethanol followed rapidly in early stage, being the highest at 72 hours (11.9%) 2) Total sugar content of the mash was decreased considerably within 48-72 hours, being 2.62% at 72 hours, and thereafter slowly. 3) Reducing sugar of the mash had a tendency of decreasing, being 0.29% at 48 hours. 4) Total acidity, volatile acidity and Formal nitrogen content of the mash were increased slowly, being 7.30, 0.20, 2.55 at 48 hours. 5) Total cells of yeast appeared the highest in 72 hours ($2.1{\times}10^8$) and thereafter decreased slowly. 6) Total cells of bacteria appeared the highest in 48 hours ($2.4{\times}10^8$) and thereafter decreased or increased slightly. 5. Takju was made from the fermented mash mixed with water to be 6% of alcohol content, and the change of alcohol content, total acidity, total cells of yeast and bateria during the storing at $30^{\circ}C$ were as follows; 1) Alcohol content of Takju was increased slightly at 24 hours (6.2%), and thereafter decreased slowly. 2) Total acidity of Takju was increased gradually, being 6.1 at 72 hours 3) Total cells of yeast and bacteria appeared the highest at 48 hours ($2.3{\times}10^8$, $1.5{\times}10^8$) and thereafter decreased slowly. 6. Alcohol content, total acidity and Formol nitrogen content of the Takju brewed with potato flour Ipkuk or wheat flour Ipkuk and steamed potatoes(1:5) were 9.8-11.3%, 5.8-7.4, 2.5-3.3 respectively, and the color of the Takju was similar to commercial Takju. 7. The results of sensory test for various experimental Takju, showed that the Takjues brewed with the materials combined with wheat flour and steamed potatoes (4:5 or 3.5:7.5) were not significantly different in color, taste and flavor from commercial Takju, However, those with potato flour and wheat flour (1:1 or 7:3) were significantly different from commercial Takju.

• 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.