• Title/Summary/Keyword: Enzyme properties

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Enzymatic Characteristics of Water-Insoluble ${\alpha}-Amylase$ Immobilized on Dithiocarbamate Wool (Dithiocarbamate Wool에 고정(固定)된 불용성(不溶性) ${\alpha}-Amylase$의 특성(特性)에 대하여)

  • Lee, Kyung-Hee;Kim, Jong-Deog;Lee, Kang-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.14 no.2
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    • pp.164-170
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    • 1985
  • Yellowish modified wool, dithiocarbamate(DTC) wool, was synthesized by partial hydrolysis in 0.2 N-NaOH reacting with carbon disulfide to use as ${\alpha}-amylase$ immobilization matrix. ${\alpha}-amylase$ was immobilized reacting with sulfide group of DTC-wool by covalent binding within 1 hour. 0.5 gram of this preparation, $DTC-wool-{\alpha}-amylase$, contained 150 ug of enzyme protein and its specific activity was about 90% of the native one. General properties of $DTC-wool-{\alpha}-amylase$ were a little different from optimum temperature, optimum pH, heat stability, kinetic constants and activation energy. An apparent Michaelis constant and maximum velocity of $DTC-wool-{\alpha}-amylase$ were 5.56 mg/ml and 0.37 mg/ml. $min^{-1}$ respectively, while activation energy was 16.6 kcal/mole.

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Immunohistochemical Study of NSE in Small Cell Lung Cancer (SCLC) Combined with Serum Assay (소세포폐암에서 Neuron Specific Enolase의 면역조직 화학염색과 혈청농도에 관한 연구)

  • Kwak, Seung-Min;Kim, Hyung-Jung;Shin, Dong-Hwan;Jang, Joong-Hyun;Lee, Hong-Lyeol;Kim, Se-Kyu;Ahn, Chul-Min;Kim, Sung-Kyu;Lee, Won-Young;Lee, Kyi-Beom
    • Tuberculosis and Respiratory Diseases
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    • v.39 no.6
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    • pp.502-510
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    • 1992
  • Background: Neuron specific enolase (NSE) is a neuronal form of the glycolytic enzyme enolase which was first found in extracts of brain tissue, and later in a variety of APUD cells and neurons of the diffuse endocrine system. SCLC shares many APUD properties with normal neuroendocrine cells. NSE immunostaining and serum NSE measurement may be a useful marker of neuroendocrine differentiation in lung tumors and diagnosis of small cell carcinoma. Methods: NSE immunohistochemical staining was done and at the same time serum NSE levels were measured in 22 small cell lung cancer and 21 non small cell lung cancer which were confirmed histologically. Results: 1) NSE immunoreactivity was detected in 9 of the 18 (50%) small cell lung cancer, in 5 of the 16 non small cell lung cancer. 2) Whereas the mean value in non-small cell lung cancer group was $11.79{\pm}4.47\;ng/ml$, the mean level of serum NSE in small cell lung cancer increased up to $59.3{\pm}77.8\;ng/ml$. In small cell lung cancer patients, mean value of limited disease group was $20.19{\pm}12.91\;ng/ml$, while mean value of extended disease group was $91.9{\pm}94.2\;ng/ml$ showing statistically significant difference. If serum levels above 20 ng/ml were tentatively defined as positive, 16 of 22 (73%) patients with SCLC had positive serum NSE level, but only one patient with NSCLC did. There was no correlation between serum NSE level and immunoreactivity of NSE. Conclusion: These studies indicate that serum NSE measurement may be a useful marker for the diagnosis and disease extent and NSE immunostaining can be used to demonstrate the neuroendocrine components of lung tumor.

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Fermentation Characteristics of Cheese Slurry prepared from Caseinates (카세인을 이용한 치즈곤죽의 발효특성)

  • Jang, Hae-Dong;Lee, Hyong-Joo
    • Korean Journal of Food Science and Technology
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    • v.17 no.5
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    • pp.389-398
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    • 1985
  • To shorten the processing of cheese slurry, four different slurries, ie, Control, Cheddar 1 and 2, and Italian-type that were made of Na-caseinates, cream, trace elements, lactic culture, and enzymes were fermented at $30^{\circ}C$ for 7days with daily stirring. PH, titratable acidity, soluble nitrogen, viable cell count, active SH groups, total volatile fatty acid, free fatty acid, electrophoretic patterns of degraded caseins, and viscosity were analyzed to investigate physicochemical properties of fermented slurries. Acid production was accelerated in the cheese slurries with protease than that without the enzyme and PH of the former was decreased after three days of fermentation to 4.90. The Change of titratable acidity agreed to PH patterns. Soluble nitrogen of the Control slurry was increased slowly for four days and then rapidly to 40% of total nitrogen while those containing protease to 70%. The protease of lactic cultures used (Streptococcus lactis and Streptococcus cremoris) broke down as-casein more rapidly than $\beta$-casein and most proteins were degraded to peptides and amino acids after three days of fermentation. Total volatile fatty acids were increased by added lipase and free fatty acids composition analyzed by GLC in cheddar slurry with 0.00001% lipase was similar to that of commercial cheddar cheese, while that in Italian-type slurry was a half of that in commercial Italian cheese. Active SH groups were increased in the cheese slurries with glutathione from fourth day of fermentation. The viscosity of slurries decreased very rapidly by addition of protease.

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Studies on Characteristics of the Cellulolytic Enzymes Produced by Pleurotus sajor-caju (Pleurotus sajor-caju가 생산(生産)하는 섬유소(纖維素) 분해(分解) 효소(酵素)의 성질(性質)에 관한 연구(硏究))

  • Hong, Jai-Sik;Lee, Ji-Yul;Kim, Dong-Han;Lyu, Gun-Sok
    • The Korean Journal of Mycology
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    • v.12 no.4
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    • pp.133-140
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    • 1984
  • Some properties of cellulolytic enzymes produced by Pleurotus sajor-caju JAFM 1017 during its growth in synthetic medium were investigated. The optimum pH of avicelase, CMCase, and ${\beta}-glucosidase$ was pH 5.5, pH 4.5 and pH 6.0, respectively. Avicelase and CMCase were stable within pH 5.0 to 6.0 and 4.0 to 6.0, respectively, and ,${\beta}-glucosidase$ was within pH 5.5 to 6.5. The optimum temperature of avicelase, CMCase and ${\beta}-glucosidase$ was the same of $40^{\circ}C$. The enzymes were stable below the optimum temperature, but the enzymes were unstable over the temperature of $50^{\circ}C$, and avicelase was losing about 91.7% of activity at $70^{\circ}C$ for 10 min. The enzyme activity of avicelase and CMCase was increased in proportion to the substrate concentration within 1% and 0.7%, respectively, and ${\beta}-glucosidase$ was within 0.1%. The Michaelis constants (Km) of avicelase and CMCase were 30.77mg avicel/ml and 14.64m Na-CMC/ml, respectively and ${\beta}-glucosidase$ was 5. 13mg salicin/ml. The reducing sugar production of avicelase was proportionaly increased until 120 min. and CMCase and ${\beta}-glucosidase$ were until 60min. The activity of three cellulolytic enzymes were increased by $Ca^{2+}$ at the concentration of $10^{-2}M$, but were inhibited by $Hg^{2+}$, $Ag^+$.

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The Radioprotective Effect of Ginseng Extracts on the liver in Mice that was irradiated by radiation (방사선이 조사된 생쥐 간에서 인삼추출물이 방사선 방어효과에 미치는 영향)

  • Ko, In-Ho;Chang, Chae-Chul;Koh, Jeong-Sam
    • Journal of radiological science and technology
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    • v.27 no.2
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    • pp.35-43
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    • 2004
  • Radioprotective effects of ginseng extracts on liver damage induced by high energy x-ray were studied. To one group of ICR male mice were given white(50 mg/kg/day for 7days, orally) and fermenta ginseng extracts(500 mg/kg/day for 7days, orally) before irrdiation. To another group were irradiated by 5 Gy dose of high energy x-ray. Contrast group were given with saline(0.1 ml). This study also investigated the radioprotective effect between SOD, CAT, hydrogen peroxide and ginseng extracts on hepatic damage. This study measured the level of superoxide dismutase(SOD), catalase(CAT), hydrogen peroxide($H_2O_2$) in liver tissue. Administrating orally white (50 mg/kg/day for 7days, orally) and fermenta ginseng extracts(500 mg/kg/day), the activity of SOD, CAT were generally increased and the hydrogen peroxide($H_2O_2$) was decreased. After irradiation, the activity of SOD, CAT were generally decreased and the hydrogen peroxide($H_2O_2$) was increased. Therefore, ginseng extracts increased antioxidative enzyme activity. And We know that the antioxidatant effect of extracts from white and fermenta ginseng protect radiation damage by direct antioxidant effect involving SOD, CAT. It was included that ginseng can protect against radiation damage through its antioxidatant properties.

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Selectivity of Bleaching Herbicides Caused by Physiological Differences between Rice and Barnyardgrass (벼와 피의 생리적(生理的) 차이(差異)에 의한 백화형(白化型) 제초제(除草劑)의 선택성(選擇性))

  • Na, J.Y.;Kim, J.S.;Kim, T.J.;Cho, K.Y.;Pyon, J.Y.
    • Korean Journal of Weed Science
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    • v.12 no.2
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    • pp.89-101
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    • 1992
  • In this study, various physiological and biochemical experiments were conducted to know whether the selectivity between rice and barnyardgrass treated with bleaching herbicides containing diphenyl ether compounds was also partly based on their basic physiological proterties such as peroxidation ability, membrane stability or antioxidant system. It seemed to be partly based on the differences of their physiological characteristics that barnyardgrass was commonly more susceptible to most of the bleaching herbicides than rice. The scenescence of intact leaf segment was more rapid in barnyardgrass than in rice, indicating that barnyardgrass is weak at early stage. Also pigment metabolic ability, antioxidant enzyme activities(peroxidase, catalase, superoxide dismutase, glutathione reductase) and antioxidant content (tocopherol, ascorbic acid, glutathione, carotenoids) were lower in barnyardgrass on the basic of fresh weight. However, lipoxygenase activity and the content of unsaturated fatty acid which is vulnerable to oxygen radicals were higher in barnyardgrass, suggesting that barnyardgrass seedling bave a properties easy to be peroxidized. The differences of PPIX (protoporphyrin IX) or carotenoid content, which are the primary substances inducing herbicide activity, were not related to the selectivity between rice and barnyardgrass.

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Peptide Properties of Rapid Salted and Fermented Anchovy Sauce Using Various Pretenses 1. Hydrolysis of Anchovy Sauce and Actomyosin by Various Pretenses (단백질 분해효소를 이용하여 제조한 속성 멸치 액젓의 펩티드 특성 1. 단백질 분해효소에 의한 멸치 액젓 및 Actomyosin의 가수분해)

  • KIM In-Soo;CHOI Young-Joon;HEU Min-Soo;CHO Young-Je;IM Yeong-Sun;GU Yeun-Suk;YEO Saeng-Gyu;PARK Jae-Woon
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.4
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    • pp.481-487
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    • 1999
  • The optimal conditions of enzymatic hydrolysis for preparation of rapid salted and fermented anchovy sauce (SFAS) using various pretenses such as trypsin, chymotrypsin, crude enzyme from squid liver and viscera, Alcalase, Neutrase and Protamex were studied. SFAS prepared with squid viscera had higher level of VBN (173.6 mg/100 g) when stored for 70 days than other samples, and peroxide values were almost equal among all samples during fermentation period. Total amino acids and nonprotein nitrogenous compounds remarkably increased as SFAS treated with Alcalase or Protamex which exhibited higher the hydrolysis rate of $57\%$ at 60 day than others. The optimal pHs of trypsin, chymotryosin, Alcalase, Neutrase and Protamex on anchovy actomyosin were 7.5, 6.5, 6.5, 7.0 and 5.0, respectively. Optimal temperatures of trypsin, chymotryosin, Alcalase and Neutrase were 55, 45, 60 and $55^{\circ}C$, respectively. Otherwise, Protamex activity increased as temperature increased from 20 to $70^{\circ}C$. Protamex had higher $K_m$ (3.545) and $V_{max}$ value (2.688) than others. Protamex affected less by NaCl had $52.5\%$ activity at the fermentation condition of $20^{\circ}C$ and $25\%$ NaCl. Protamex appeared to be very effective for the hydrolysis of crude actomyosin from ancnovy.

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Effects of Added WPC and WP on the Quality and Shelf Life of Tofu (WPC 및 WP 첨가가 두부 품질 및 저장성에 미치는 영향)

  • Kim, Jong-Un;Song, Kwang-Young;Seo, Kun-Ho;Yoon, Yoh-Chang
    • Journal of Dairy Science and Biotechnology
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    • v.30 no.2
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    • pp.93-109
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    • 2012
  • This study was performed to investigate the effects of added whey protein concentrates (WPC) and whey powder (WP) on the quality and shelf life of Tofu, a traditional food in Korea. Combined whey powder and whey protein concentrates were obtained at drainage after the casein was separated by using rennet enzyme or acidification of milk. We manufactured whey Tofu and evaluated its nutritional quality by testing, the general composition for yield, moisture, pH, crude protein, crude fat, carbohydrate, rheology, sensory properties, and change during storage. 1. The general compositions of WPC and WP were as follows: (a) WPC: moisture, 5.9%; crude protein, 56.2%; crude fat, 0.1%; carbohydrate, 32.6%; ash, 5.2%; and pH 5.93 and (b) WP: moisture, 3.7%; crude protein, 13.2%; crude fat, 1.6%; carbohydrate, 74.4%; ash, 7.1%; and pH, 6.65. 2. The yield of Tofu was as follows: (a) in WPC, the content was $CaCl_2$:GDL=6:4 > $CaCl_2$:GDL=9:1 > $CaCl_2$:GDL=7:3 > $CaCl_2$:GDL=8:2 and (b) in WP, 2% addition was the highest (265%) at $13.3g/cm^2$, but with 4% addition WP was the lowest (184%) at $22.2g/cm^2$. 3. The moisture content of Tofu was as follows: (a) in WPC, the content was $CaCl_2$:GDL = 6:4 > $CaCl_2$:GDL=9:1 > $CaCl_2$:GDL=7:3 > $CaCl_2$:GDL=8:2 and (b) in WP, 2% addition was the highest at 79.82% ($13.3g/cm^2$), but 4% was the lowest at 75.18% ($22.2g/cm^2$). 4. The pH of Tofu was as follows: (a) in WPC, the value was WPC 6% > WPC 4% > WPC 2% > control and $CaCl_2$:GDL=6:4 > $CaCl_2$:GDL=8:2 > $CaCl_2$:GDL=9:1 > $CaCl_2$:GDL=7:3 and (b) in WP, WP 4% > WP 2% > control. 5. The ash content of Tofu was as follows: (a) in WPC, the content was $CaCl_2$:GDL=8:2 > $CaCl_2$:GDL=7:3 > $CaCl_2$:GDL=6:4 > $CaCl_2$:GDL=9:1 and (b) in WP, there was no difference between 2% and 4% addition. 6. The crude protein content of Tofu was as follows: (a) in WPC, the content was $CaCl_2$:GDL=8:2 > $CaCl_2$:GDL=7:3 > $CaCl_2$:GDL=9:1 > $CaCl_2$:GDL=6:4 and (b) in WP, there was no difference between 2% and 4% addition. 7. The crude fat content of Tofu was as follows: (a) in WPC, the content was $CaCl_2$:GDL=8:2 > $CaCl_2$:GDL=7:3 > $CaCl_2$:GDL=9:1 > $CaCl_2$:GDL=6:4 and (b) in WP, values decreased with increasing pressed weight. 8. The carbohydrate content of Tofu was as follows: (a) in WPC, the content was $CaCl_2$:GDL=8:2 > $CaCl_2$:GDL=7:3 > $CaCl_2$:GDL=6:4 > $CaCl_2$:GDL=9:1 and (b) in WP, values increased with increasing pressed weight. 9. The rheology test results of Tofu were as follows: (a) in WPC, hardness and brittleness was highest with $CaCl_2$:GDL=8:2 and 6% added WPC. Cohesiveness was highest with $CaCl_2$:GDL=6:4 and 2% added WPC. Elasticity was the highest with $CaCl_2$:GDL=7:3 and the added WPC control. (b) in WP, hardness was the highest with $22.2g/cm^2$ and added WP control. Cohesiveness was the highest with $17.8g/cm^2$ and added WP 2%. Elasticity was the highest with $17.8g/cm^2$ and added WP 4%. Brittleness was the highest with $17.8g/cm^2$ and added WP control. 10. The sensory test results of Tofu were as follows: (a) in WPC, the texture, flavor, color, and smell were the highest with $CaCl_2$:GDL=6:4 and 6% added WPC. (b) in WP, the texture was the highest in the control with $22.2g/cm^2$. Flavor and smell were the highest in WP 2% and $22.2g/cm^2$. Color was the highest in WP 2% and $17.8g/cm^2$. 11. The quality change of Tofu during storage was as follows: (a) in WPC, after 60 h, all samples began to get spoiled and their color changed, and mold began to germinate. (b) in WP, the result was similar, but the rate of spoilage was more rapid than that in the control.

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Sesquiterpenoids Bioconversion Analysis by Wood Rot Fungi

  • Lee, Su-Yeon;Ryu, Sun-Hwa;Choi, In-Gyu;Kim, Myungkil
    • 한국균학회소식:학술대회논문집
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    • 2016.05a
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    • pp.19-20
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    • 2016
  • Sesquiterpenoids are defined as $C_{15}$ compounds derived from farnesyl pyrophosphate (FPP), and their complex structures are found in the tissue of many diverse plants (Degenhardt et al. 2009). FPP's long chain length and additional double bond enables its conversion to a huge range of mono-, di-, and tri-cyclic structures. A number of cyclic sesquiterpenes with alcohol, aldehyde, and ketone derivatives have key biological and medicinal properties (Fraga 1999). Fungi, such as the wood-rotting Polyporus brumalis, are excellent sources of pharmaceutically interesting natural products such as sesquiterpenoids. In this study, we investigated the biosynthesis of P. brumalis sesquiterpenoids on modified medium. Fungal suspensions of 11 white rot species were inoculated in modified medium containing $C_6H_{12}O_6$, $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ for 20 days. Cultivation was stopped by solvent extraction via separation of the mycelium. The metabolites were identified as follows: propionic acid (1), mevalonic acid lactone (2), ${\beta}$-eudesmane (3), and ${\beta}$-eudesmol (4), respectively (Figure 1). The main peaks of ${\beta}$-eudesmane and ${\beta}$-eudesmol, which were indicative of sesquiterpene structures, were consistently detected for 5, 7, 12, and 15 days These results demonstrated the existence of terpene metabolism in the mycelium of P. brumalis. Polyporus spp. are known to generate flavor components such as methyl 2,4-dihydroxy-3,6-dimethyl benzoate; 2-hydroxy-4-methoxy-6-methyl benzoic acid; 3-hydroxy-5-methyl phenol; and 3-methoxy-2,5-dimethyl phenol in submerged cultures (Hoffmann and Esser 1978). Drimanes of sesquiterpenes were reported as metabolites from P. arcularius and shown to exhibit antimicrobial activity against Gram-positive bacteria such as Staphylococcus aureus (Fleck et al. 1996). The main metabolites of P. brumalis, ${\beta}$-Eudesmol and ${\beta}$-eudesmane, were categorized as eudesmane-type sesquiterpene structures. The eudesmane skeleton could be biosynthesized from FPP-derived IPP, and approximately 1,000 structures have been identified in plants as essential oils. The biosynthesis of eudesmol from P. brumalis may thus be an important tool for the production of useful natural compounds as presumed from its identified potent bioactivity in plants. Essential oils comprising eudesmane-type sesquiterpenoids have been previously and extensively researched (Wu et al. 2006). ${\beta}$-Eudesmol is a well-known and important eudesmane alcohol with an anticholinergic effect in the vascular endothelium (Tsuneki et al. 2005). Additionally, recent studies demonstrated that ${\beta}$-eudesmol acts as a channel blocker for nicotinic acetylcholine receptors at the neuromuscular junction, and it can inhibit angiogenesis in vitro and in vivo by blocking the mitogen-activated protein kinase (MAPK) signaling pathway (Seo et al. 2011). Variation of nutrients was conducted to determine an optimum condition for the biosynthesis of sesquiterpenes by P. brumalis. Genes encoding terpene synthases, which are crucial to the terpene synthesis pathway, generally respond to environmental factors such as pH, temperature, and available nutrients (Hoffmeister and Keller 2007, Yu and Keller 2005). Calvo et al. described the effect of major nutrients, carbon and nitrogen, on the synthesis of secondary metabolites (Calvo et al. 2002). P. brumalis did not prefer to synthesize sesquiterpenes under all growth conditions. Results of differences in metabolites observed in P. brumalis grown in PDB and modified medium highlighted the potential effect inorganic sources such as $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ on sesquiterpene synthesis. ${\beta}$-eudesmol was apparent during cultivation except for when P. brumalis was grown on $MgSO_4$-free medium. These results demonstrated that $MgSO_4$ can specifically control the biosynthesis of ${\beta}$-eudesmol. Magnesium has been reported as a cofactor that binds to sesquiterpene synthase (Agger et al. 2008). Specifically, the $Mg^{2+}$ ions bind to two conserved metal-binding motifs. These metal ions complex to the substrate pyrophosphate, thereby promoting the ionization of the leaving groups of FPP and resulting in the generation of a highly reactive allylic cation. Effect of magnesium source on the sesquiterpene biosynthesis was also identified via analysis of the concentration of total carbohydrates. Our current study offered further insight that fungal sesquiterpene biosynthesis can be controlled by nutrients. To profile the metabolites of P. brumalis, the cultures were extracted based on the growth curve. Despite metabolites produced during mycelia growth, there was difficulty in detecting significant changes in metabolite production, especially those at low concentrations. These compounds may be of interest in understanding their synthetic mechanisms in P. brumalis. The synthesis of terpene compounds began during the growth phase at day 9. Sesquiterpene synthesis occurred after growth was complete. At day 9, drimenol, farnesol, and mevalonic lactone (or mevalonic acid lactone) were identified. Mevalonic acid lactone is the precursor of the mevalonic pathway, and particularly, it is a precursor for a number of biologically important lipids, including cholesterol hormones (Buckley et al. 2002). Farnesol is the precursor of sesquiterpenoids. Drimenol compounds, bi-cyclic-sesquiterpene alcohols, can be synthesized from trans-trans farnesol via cyclization and rearrangement (Polovinka et al. 1994). They have also been identified in the basidiomycota Lentinus lepideus as secondary metabolites. After 12 days in the growth phase, ${\beta}$-elemene caryophyllene, ${\delta}$-cadiene, and eudesmane were detected with ${\beta}$-eudesmol. The data showed the synthesis of sesquiterpene hydrocarbons with bi-cyclic structures. These compounds can be synthesized from FPP by cyclization. Cyclic terpenoids are synthesized through the formation of a carbon skeleton from linear precursors by terpene cyclase, which is followed by chemical modification by oxidation, reduction, methylation, etc. Sesquiterpene cyclase is a key branch-point enzyme that catalyzes the complex intermolecular cyclization of the linear prenyl diphosphate into cyclic hydrocarbons (Toyomasu et al. 2007). After 20 days in stationary phase, the oxygenated structures eudesmol, elemol, and caryophyllene oxide were detected. Thus, after growth, sesquiterpenes were identified. Per these results, we showed that terpene metabolism in wood-rotting fungi occurs in the stationary phase. We also showed that such metabolism can be controlled by magnesium supplementation in the growth medium. In conclusion, we identified P. brumalis as a wood-rotting fungus that can produce sesquiterpenes. To mechanistically understand eudesmane-type sesquiterpene biosynthesis in P. brumalis, further research into the genes regulating the dynamics of such biosynthesis is warranted.

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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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