• Title/Summary/Keyword: mannitol production

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Isolation and Characterization of Dextrans Produced by Leuconostoc sp. strain JYY4 from Fermented Kimchi

  • Gu, Ji-Joong;Ha, Yoo-Jin;Yoo, Sun-Kyun
    • Journal of the Korean Applied Science and Technology
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    • v.32 no.4
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    • pp.758-766
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    • 2015
  • Dextran is a generic term for a bacterial exopolysaccharide synthesized from sucrose and composed of chains of D-glucose units connected by ${\alpha}$-1,6-linkages by using dextransucrases. Dextran could be used as vicosifying, stabilizing, emulsifying, gelling, bulking, dietary fiber, prebiotics, and water holding agents. We isolated new strain capable of producing dextran from Korean traditional kimchi and identified as Leuconostoc sp. strain JYY4. Batch fermentation was conducted in bioreactor with a working volume of 3 L. The media was MMY and 15% (w/v) sucrose. Mineral medium consisted of $3.0g\;KH_2PO_4$, $0.01g\;FeSO_4$, $H_2O$, $0.01g\;MnSO_4$, $4H_2O$, $0.2g\;MgSO_4\;7H_2O$, 0.01 g NaCl, $0.05g\;CaCl_2$ per 1 liter deionized water. The pH of media was initially adjusted to 6.0. The inoculation rate was 1.0% (v/v) of the working volume. Temperature was maintained at $28^{\circ}C$. The agitation rate was 100 rpm. The production pattern of dextran was associated with the cell growth. After 24 hr dextran reached its highest concentration of 59.4 g/L. The sucrose was consumed completely after 40 hr. Growth reached stationery phase when sucrose became limiting, regardless of the presence of fructose or mannitol. When the specific growth rate was 0.54 hr-1, utilization averaged 5.8 g/L-hr. The yield and productivity of dextran were 80% and 2.0 g/L-hr, respectively. Dextrans produced by were separated to two different size by an alcohol fraction method. The size of high molecular weight dextran (45% alcohol, v/v), less soluble dextran, was between MW 500,000 and 2,000,000. Soluble dextran (55% alcohol, v/v) was between 70,000 and 150,000. The molecular weight average of total dextran (70% alcohol, v/v) was between 150,000 to 500,000. The enzymatic hydrolyzates of total dextran of ATCC 13146 showed branched dextrans by Penicillium dextranase contained of glucose, isomaltose, isomaltotriose, and isomaltooligosaccharides greater than DP4 (degree of polymerization) that had branch points. Compounds greater than DP4 were branched isomaltooligosaacharides. Hydrolysates by the Lipomyces dextranase produced the same composition of oligosaccharides as those by Penicillin dextranase.

Antioxidant Favors the Developmental Competence of Porcine Parthenogenotes by Reducing Reactive Oxygen Species

  • Hossein, Mohammad Shamim;Kim, Yeun Wook;Park, Seon Mi;Koo, Ok Jae;Hashem, Md Abul;Bhandari, Dilip P;Jeong, Yeon Woo;Kim, Sue;Kim, Ji Hye;Lee, Eu Gine;Park, Sun Woo;Kang, Sung Keun;Lee, Byeong Chun;Hwang, Woo Suk
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.3
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    • pp.334-339
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    • 2007
  • Reactive oxygen species (ROS) generate during electrical activation of oocytes which has detrimental effects on embryo survival when overwhelmed. The present study was designed to investigate the ability of L-ascorbic acid, a novel water soluble antioxidant, to reduce the ROS level in developing embryos and their subsequent effects on embryo development in vitro. The compact cumulus oocyte complexes (COCs) were cultured in tissue culture medium (TCM)-199 supplemented with 10 ng/ml epidermal growth factor, 4 IU/ml pregnant mare serum gonadotropin (PMSG), and human chorionic gonadotropin (hCG) and 10% (v/v) porcine follicular fluid (pFF) for 44 h. After maturation culture, the denuded oocytes were activated with a single DC pulse of 2.0 kV/cm in 0.3 M mannitol solution containing 0.5 mM of HEPES, 0.1 mM of $CaCl_2$ and 0.1 mM of $MgCl_2$ for $30{\mu}s$ using a BTX Electro-cell Manipulator. The activated oocytes were cultured in modified North Carolina State University-23 (mNSCU-23) medium for 168 h. The level of $H_2O_2$ in each embryo was measured by the dichlorohydrofluorescein diacetate (DCHFDA) method at 48 h after activation. The blastocyst formation rate was significantly higher when culture medium was supplemented with 50 and $100{\mu}M$ L-ascorbic acid (31.2 and 38.7%, respectively) compared to non-supplemented (16.1%) group. Accordingly, significantly more cells in blastocyst were found for 50 and $100{\mu}M$ L-ascorbic acid (50.0 and 56.4, respectively) compared to 0 and $200{\mu}M$ L-ascorbic acid (36.5 and 39.8, respectively). L-ascorbic acid reduces the $H_2O_2$ level in developing embryos in a dose-dependant manner. The $H_2O_2$ level (pixels/ embryos) was 191.5, 141.0, 124.0 and 163.3 for 0, 50, 100 and $200{\mu}M$ L-ascorbic acid, respectively. So, we recommend to supplement 50 or $100{\mu}M$ L-ascorbic acid in porcine in vitro culture medium.

Production of cloned Mice by Nuclear Transplantation and Electrofusion Using 2- or 8-Cell Stage Mouse Embryo as Nuclear Donor (2- 및 8- 세포기 생쥐 수정란의 핵이식 및 전기융합법에 의한 복제산자의 생산)

  • 박준규;조성근;박희성;박충생
    • Journal of Embryo Transfer
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    • v.10 no.3
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    • pp.209-217
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    • 1995
  • The present study was carried out to develop a cloning technology of mouse embryos by nuclear transplantation with electrofusion and to produce cloned offsprings by transfer of reconstituted embryos. A single nucleus from two- and eight-cell embryos was transplanted into the enucleated two-cell embryos by rnicromanipulation. The fusion of nucleus with recipient cytoplasm and the subsequent development of reconstituted embryos in vitro as well as in vivo to term were examined to determine the optimal electrofusion parameters for nuclear transplantation in mouse embryos. The successful enucleation of donor embryos was 84.9 and 83.3% in two- and eight-cell stage, respectively, and the successful injection of nucleus from two- and eight-cell donor embryos into the perivitelline space of enucleated two-cell embryos were 85.1 and 84.7%, respectively. No significant differences were found in enucleation or injection rate between the cell stages of donor embryos. When the blastomeres of intact two-cell mouse embryos were electrofused in 0.3 M mannitol medium(100 $\mu$sec., 3 pulses), the fusion rate was similarly 93.2, 92.2 and 92.0% in 1.0, 1.5 and 2.0 kV /crn, respectively, but in vitro development to blastocyst of the fused two-cell embryos was significantly(P<0.05) lower in 2.0 kV/cm (63.4%) than in 1.0 kV/cm (91.7%) or 1.5 kV/cm (82.4%). The development in vitro to eight-cell stage of the reconstituted embryos with nucleus from two-cell stage(45.5%) was significantly(P<0.05) higher than that from eight-cell stage blastomeres (16.7%). The number of blastomeres of the intact embryos at blastocyst stage was 50i0.6 and 55$\pm$2.4 in in vitro and in vivo cultured mouse embryos, respectively, but significantly(P<0.05) decreased to 35$\pm$0.7 in nuclear transplanted blastocyst embryos. The conception rate of mice following embryo transfer was 32.1% in the reconstituted two-cell embryos using two-cell donor nuclei, which was comparable to the fresh two-cell embryos(40.6%). However, the rate of development in vivo to term following embryo transfer of the reconstituted two-cell embryos using two-cell donor nuclei (23.5%) was significantly(P<0.05) lower compared with the percentage of two-cell fresh embryos(31.5%).

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Quality Characteristics of Kimchi according to Garlic Content during Fermentation (마늘 첨가량에 따른 김치의 발효 중 품질 특성 변화)

  • Choi, Yun-Jeong;Hwang, Ye-Seul;Hong, Sung Wook;Lee, Mi-Ai
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.11
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    • pp.1638-1648
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    • 2016
  • This study was conducted to analyze the quality characteristics of kimchi in terms of garlic content (0~4.5%). Kimchi was made at $4^{\circ}C$ for 8 weeks, and pH, acidity, organic acid content, free sugar content, microbial counts, flavor pattern, and sensory characteristics were measured. The results show that kimchi containing garlic had a higher pH and lower acidity during fermentation than control kimchi without garlic. Principal component analysis enabled differentiation of the flavor pattern of kimchi according to fermentation period and garlic content. Addition of garlic to kimchi significantly decreased the numbers of total bacteria and lactic acid bacteria for 2 weeks after production. The numbers of total bacteria and lactic acid bacteria increased rapidly up to 2 weeks during fermentation and thereafter decreased gradually. Coliform counts were higher in the control than in kimchi containing garlic, whereas there was no detection after 4 weeks. Yeast and mold counts decreased significantly with increasing garlic content during the initial fermentation stage. Counts could not be detected in kimchi containing garlic. After 4 weeks, counts could not be detected in kimchi without garlic. Among kimchi with different garlic contents, fermentation was slower in kimchi with high garlic content; scores for off-odor and off-note taste were lower as well.

The Effect of Oocyte Activation on Development of Porcine Cloned Embryos

  • Kim, Y.S.;Lee, S. L.;Park, G. J.;S. Y. Choe
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.124-124
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    • 2003
  • The successful development of embryos cloned by nuclear transfer (NT)have been dependent on a wide range of known factors including cell cycle of donor and recipient ooplast, oocyte quality, NT procedure and oocyte activation. The present study compared the development of cloned porcine embryos following different activation treatments. Cumulus-oocyte complexes (COCs) were aspirated from 26 mm follicles of slaughterhouse ovaries and cultured for 22 h in NCSU #23 medium supplemented with 10% porcine follicular fluid, 0.57 mM cysteine, 0.5 g/mL LH, 0.5 g/mL FSH and 10 ng/mL EGF. The COCs were further cultured for an additional 22 h in the same medium at $39{\cird}C$ in an atmosphere of 5% $CO_2$ in air, without hormonal supplements. Primary cultures of fibroblasts isolated from a female fetus on day 40 of gestation were established in DMEM + 15% FCS. For nuclear donation, cells at the 5th-6th passage were cultured in DMEM +0.5% FCS for 5 days in order to arrest the cells in G0/Gl. After enucleation, oocytes were reconstructed by transfer of donor cells and fusion with three DC pulses (1.4 KV/cm, 30 sec) in 0.28 M mannitol containing 0.01 mM $CaCl_2$ and 0.01 mM $MgCl_2$. Eggs were then divided into three treatment groups, control (without further treatment, Group 1), eggs cultured in 10 g/ml cycloheximide (CHX) for 5 h (Group 2), and eggs cultured in 1.9 mM 6-dimethylaminopurine (6-DMAP) for 5 h (Group 3). The eggs were then cultured in sets of 30 in 60 I drops of NCSU#23 supplemented with 4mg/ml BSA (essentially fatty acid free) until day 7 at $39{\circ}C$ in a humidified atmosphere of 5% $CO_2$. On day 4 the culture were fed by adding 20 I NCSU #23 supplemented with 10% FBS. Development rates into blastocysts were significantly higher (P<0.05) in Group 3 embryos compared to Group 1 controls ($27.6 \mu 2.7% vs. 20.1 \mu 4.1%$, respectively), but rates did not differ in Group 2 compared to control ($23.8 \mu 5.7%$). Total cell number in Group 3 blastocysts was however significantly higher (P<0.05) than in Groups 1 and 2 ($44.6 \mu 2.4 vs. 19.9 \mu 1.9 and 21.9 \mu 2.1$, respectively). These results suggest that 6-DMAP is more efficient than cycloheximide in the activation of electrically fused NT oocytes during in vitro production of cloned porcine embryos.

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Properties of the Silkworm (Bombyx mori) Dongchunghacho, a Newly Developed Korean Medicinal Insect-borne Mushroom: Mass-production and Pharmacological Actions (한국에서 개발된 곤충유래 약용버섯인 누에동충하초의 생산기술개발 및 약리학적 특성)

  • Lee, Sang Mong;Kim, Yong Gyun;Park, Hyean Cheal;Kim, Keun Ki;Son, Hong Joo;Hong, Chang Oh;Park, Nam Sook
    • Journal of Life Science
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    • v.27 no.2
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    • pp.247-266
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    • 2017
  • Cordyceps is a traditional Chinese medicinal herb well-known in China, Korea and Japan since B.C. 2,000. The original entomopathogenic fungus, Cordyceps sinensis belonging to the genus Cordyceps could not be found inside Korean peninsula due to the absence of the host insect for the corresponding entomogenous fungus. The development of artificial production methods of Korean type Cordyceps using the silkworm Bombyx mori as in vivo culture medium for the the entomopathogenic fungus Paecilomyces tenuipes is the first, and wonderful occasion in the research history of insect industry of this global world. The aim of this article is to review the historical research background, mass-production methods, and pharmacological effects of the silkworm-dongchunghacho (Paecilomyces tenuipes) which is a newly developed Korean medicinal insect-borne mushroom, and another non-insect-borne medicinal mushroom (Cordyceps militaris and Cordyceps pruinosa). Their biological actions include anti-tumor, immunostimulating, anti-fatigue, anti-stress, anti-oxidant, anti-aging, anti-diabetic, anti-inflammatory, anti-thrombosis, hypolipidaemic and insecticidal effects. The bioactive principles are protein-bound polysaccharides (hexose, hexosamin), cordycepin, D-manitol, acidic polysaccharide etc. Protein-bound polysaccharides and n-butanol fractions were demonstrated to show a significant anti-tumor activities but did not show a cytotoxicities. D-mannitol exhibited a significant prolongation of the life span in tumor bearing mice. Ergosterol did not show an efficient anti-tumor activity, but showed a significant phagocytosis enhancing activity. Anti-tumor activity of silkworm-dongchunghacho might be attributed to immuno-stimulating activities rather than cytotoxic effects [164]. Also this review comprises the breeding of Dongchunghacho varieties, optimization of culture conditions, improvement of learning and memory by Dongchunghacho, application of them as foods and chemical constituents.

한국인으로부터 분리한 비피더스균의 특성과 Bifidobacterium longum A-2의 임상실험에 관한 연구

  • Kim, Yeong-Chan
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 1998.10a
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    • pp.83-106
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    • 1998
  • This study was conducted to investigate the probiotics(acid and bile resistance, fermentation properties, viability, cholesterol assimilation, antimicrobial activity, antimutagenicity, and immunoactivation) of the strains of bifidobacteria isolated from healthy Koreans and to investigate the effects of oral administration of Bifidobacterium longum A-2 on the fecal microflora, ${\beta}-glucuronidase$ activity, pH values, Ammonia concentration. The experimental results are summarized as follows: The probiotics were tested for 23 strains including three commer챠al strains as controls. Compared to other strains, strains of A-2 and A-9 showed more acid resistance whereas A-2, A-5, A-13, A-14, A-18 and A-22 showed excellent bile resistances. The properties of bifidobacteria during fermentation were tested. Strains of A-1, A-2, A-3, A-4, A-6, and A-23 resulted in less than pH 4.5 and titratable acidity over 0.90 after 24 hr of fermentation. When the strains of A-2 was grown with glucose, maltose, and fructooligosaccharide, the acetic acid production were higher than with sorbitol and mannitol. The storage stability of the strains of A-2 and A-22 were tesed, indicating the strain A-2 was more stable over 10 days of storage at both $4^{\circ}C$ and $20^{\circ}C$ than A-22. The strains of A-8, A-10, A-11, A-12 and A-20 assimilated more than 30% of cholesterol included in the media. The strains of A-1 and A-2 showed antimicrobial activity against Sta. aureus. The antimutagenicity of the strains were also tested, showing that the mutation was suppressed more by three strains(A-2, A-12, and A-23). In addition, strain A-5 improved immunological activity(phagocytosis, $TNF-{\alpha}$, IL-6) more than other strains. In the effects of oral administration of Bif. longum A-2, the number of fecal bifidobacteria was siginificantly increased(p<0.01) and the level of fecal ${\beta}-glucuronidase$ also was siginificantly reduced(p<0.05). However there were no siginificant differences in the level of Lαctobacilli, Enterobacteriaceae, Clostridium perfringens, pH and ammonia by the administration. The results suggested that Bif. longum A-2 may be met the criteria for probiotics culture.

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Effects of High Glucose and Advanced Glycosylation Endproducts(AGE) on the in vitro Permeability Model (당과 후기당화합물의 생체 외 사구체여과율 모델에 대한 역할)

  • Lee Jun-Ho;Ha Tae-Sun
    • Childhood Kidney Diseases
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    • v.10 no.1
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    • pp.8-17
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    • 2006
  • Purpose : We describe the changes of rat glomerular epithelial cells when exposed to high levels of glucose and advanced glycosylation endproducts(AGE) in the in vitro diabetic condition. We expect morphological alteration of glomerular epithelial cells and permeability changes experimentally and we may correlate the results with a mechanism of proteinuria in DM. Methods : We made 0.2 M glucose-6-phsphate solution mixed with PBS(pH 7.4) containing 50 mg/mL BSA and pretense inhibitor for preparation of AGE. As control, we used BSA. We manufactured and symbolized five culture dishes as follows; B5 - normal glucose(5 mM) + BSA, B30 - high glucose(30 mM) + BSA, A5 - normal glucose(5 mM) + AGE, A30 - high glucose(30 mM) + AGE, A/B 25 - normal glucose(5 mM) + 25 mM of mannitol(osmotic control). After the incubation period of both two days and seven days, we measured the amount of heparan sulfate proteoglycan(HSPG) in each dish by ELISA and compared them with the B5 dish at 2nd and 7th incubation days. We observed the morphological changes of epithelial cells in each culture dish using scanning electron microscopy(SEM). We tried the permeability assay of glomerular epithelial cells using cellulose semi-permeable membrane measuring the amount of filtered BSA through the apical chamber for 2 hours by sandwich ELISA. Results : On the 2nd incubation day, there was no significant difference in the amount of HSPG between the 5 culture dishes. But on the 7th incubation day, the amount of HSPG increased by 10% compared with the B5 dish on the 2nd day except the A30 dish(P<0.05). Compared with the B5 dish on the 7th day the amount of HSPG in A30 and B30 dish decreased to 77.8% and 95.3% of baseline, respectively(P>0.05). In the osmotic control group (A/B 25) no significant correlation was observed. On the SEM, we could see the separated intercellular junction and fused microvilli of glomerular epithelial cells in the culture dishes where AGE was added. The permeability of BSA increased by 19% only in the A30 dish on the 7th day compared with B5 dish on the 7th day in the permeability assay(P<0.05). Conclusion: We observed not only the role of a high level of glucose and AGE in decreasing the production of HSPG of glomerular epithelial cells in vitro, but also their additive effect. However, the role of AGE is greater than that of glucose. These results seems to correlate with the defects in charge selective barrier. Morphological changes of the disruption of intercellular junction and fused microvilli of glomerular epithelial cells seem to correlate with the defects in size-selective barrier. Therefore, we can explain the increased permeability of glomerular epithelial units in the in vitro diabetic condition.

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Biological Activities and Artificial Cultivation of Cordyceps pruinosa Petch (붉은자루동충하초의 자실체 증식 특성)

  • Hong, In-Pyo;Nam, Sung-Hee;Jung, I-Yeon;Sung, Gyoo-Byung;Kim, Hyun-Bok;Jung, In-Mo;Cho, Soo-Muk;Lee, Min-Woong
    • Journal of Mushroom
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    • v.3 no.4
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    • pp.133-139
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    • 2005
  • Fruiting bodies of Cordyceps have been regarded as popular folk and effective medicines to treat human diseases such as asthma, bronchial and lung inflammation, and kidney disease. Cordyceps pruinosa (Clavicipitaceae; Hypocreales; Ascomycotina) has received special attention for medicinal purpose due to its various physiological activites. The nucleoside derivative N6-(2-hydroxyethyl) adenosine (HEA) isolated from it showed a $Ca^{2+}$ antagonistic effect and negative inotropic response. The artificial production of fruiting body of C. pruinosa has not been tried successfully yet by using living silkworm substrate. To develop techniques for the production of C. pruinosa stromata on a large scale, the infection of Bombyx mori with C. pruinosa and the growth characteristics of stroma of C. pruinosa were investigated. Also, studied about biological activities of fruiting body formed on silkworm. Infection rate of the silkworm pupae with C. pruinosa was the highest in injection inoculation. The formation of the fruiting body of C. pruinosa was quite good in the room controlled at $21{\sim}25^{\circ}C$, over 91% of relative humidity and over 1500 lx. Glucose concentration was high in the fruiting bodies of the silkworm pupae infected with C. pruinosa on a dry weight basis. The most abundant amino acid in the fruiting bodies was arginine and phenylalanine. The fruiting bodies of silkworm pupae infected with C. pruinosa was rich in oleic acid. The high amount of citric acid was found in the fruiting bodies of silkworm pupae infected with C. pruinosa.

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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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