• Title/Summary/Keyword: 9K 배지

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Haemophilus K-12균주로부터 황산전이효소의 분리정제

  • 김동현;김병택;이남수
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.04a
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    • pp.257-257
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    • 1994
  • 생쥐의 장내세균으로 부터 황산전이 반응을 촉매하는 효소인 sulfotransferase를 생산하는 균주를 분리하였으며 동정결과 Haemophilus속 균주로 확인되어 Haemophilus K-12라 명명하였다. 균의 성장과 효소활성과의 관계를 보면 균은 10시간에서 완전히 성장하였으며 효소활성도 이와 비례하였다. Haemophilus K-12의 배지조성에 따른 sulfotransferase의 생산성을 Brain Heart Infusion 배지에서의 생산성과 비교해보면 탄소원으로는 sucrose가 0.2%농도에서 584%로 가장 좋았으며 질소원으로는 yeast extract가 266%로 가장 좋았다. 공여체로 PNS를 최종농도 1mM로 하여 배지에 첨가하였을때 212%로 가장 높은 효소증가를 보였다. 2가금속이온에 의한 효소증가는 현저하지 않았으며 $Mn^{2+}$이 107%로 가장 높았고$Zn^{2+}$와 EDTA에 의해서는 저해를 받았다. 이상의 결과를 종합하여 균배양을 위한 이상적인 배지조성을 sucrose 0.2%, yeast extract 1%, $Na_2$HPO$_4$ 0.25%, NaCl 0.5%로 결정하였다. 결정된 최적배지에 균을 10L 배양하여 초음파 처리, 원심분리한 것을 70 % ammonium sulfate fractionation, DEAE-cellulose column chromatography를 2회, Hydroxyapatite column chromatography, chromatdfocusing column chromatography, Silica PAE column chromatography, Sephacryl S-300 superfine column chromatography를 행한결과 specific activity가 6.76 umoie/min/mg protein인 효소액을 얻었으며 homogeneous enzyme였다. 이렇게 해서 얻은 효소를 이용하여 수용체 기질 특이성을 측정한 결과 1-naphthol이 phenol을 100%로 하였을 때 233%로 가장 좋았으며 Eubacterium A-44 sulfotransferase의 좋은 기질이었던 p-acetaminophenol, tyramine, 9-phenanthrol은 좋은 기질이 되지 못했다. 이상의 결과로 미루어 보아Haemophilus K-12 sulfotransferase는 지금까지 보고된 bacterial sulfotrasferase와는 다른 효소로 사려되며 효소반응기전의 규명이 이루어지면 산업적 응용이 가능할것으로 사료된다.

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Development of New Mushroom Substrate using Kapok Seedcake for Bottle Culture of Oyster Mushroom(Pleurotus ostreatus). (케이폭박을 이용한 병재배 느타리버섯의 대체배지 개발)

  • Won, Seon-Yi;Lee, Yun-Hae;Jeon, Dae-Hoon;Ju, Young-Cheoul;Lee, Yong-Beom
    • The Korean Journal of Mycology
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    • v.38 no.2
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    • pp.130-135
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    • 2010
  • To select the viable alternative substrates among the variable organic substrates for productivity enhancement and production cost-reduction of oyster mushroom in bottle culture, this study was carried out at mushroom research institute of GGRDA in 2007. In bottle culture of oyster mushroom (Plerutus ostreatus), the seedcakes of rape (RS), soybean (SS), coconut (CCS), and kapok (KS) were examined as substitute of cotton seedcake which was primary nutritive material of mushroom growing substrate. The chemical properties of substrate mixed with kapok seedcake is similar to the mixture with cotton seedcake in T-C, T-N, C/N ratio, and other nutrients. Mixed growing substrate containing cotton seedcake and kapok seedcake was superior to other mixtures 99.2% and 99.5%, respectively in spawning ratio and was faster mycellium growth in column test than that of soybeen seedcake, cotton + soybeen seedcake, and coconut seedcake. The period required in first pin-heading was 1-2 days longer in rape and soybeen seedcake mixture. Also there wad no primodia and fruitbody formation at soybeen seedcake mixture which had highest T-N content among the other mixed substrates. Yield per bottle and biological efficiency were highest of 144.6 g and 75.4%, respectively at kapok seedcake mixture. As a result, this study found that cotton seedcake can be replaced with kapok seedcake in bottle culture of oyster mushroom.

Cultivation of Oyster Mushroom[Pleurotus ostreatus (Jacq. ex Fr.) Kummer] at Medium with Ca(OH)2 Treatment (수산화칼슘[Ca(OH)2] 처리 배지를 이용한 느타리버섯의 재배)

  • Baek, Seung-Hwa;Lee, Yeong-Il;Yoo, Ki-Yong;Han, Seong-Soo
    • Korean Journal of Environmental Agriculture
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    • v.27 no.4
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    • pp.303-313
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    • 2008
  • This research is aimed to investigate the cultivation method of oyster mushroom[Pleurotus ostreatus (Jacq. ex Fr.) Kummer] at the non-sterilized medium with $Ca(OH)_2$ treatment. Therefore, experiments were carried out to develop non-sterilization method of medium by addition of $Ca(OH)_2$ for omission of heat sterilization progress of medium. General components, minerals and amino acid in Jiri wild type No. 1 (Pleurotus ostreatus) and production cost were analyzed. For the purpose of omission of heat sterilization progress, treatment ratio of $Ca(OH)_2$ (purity 95%) was 5%(w/w) of dry medium. Initial pH of this medium was 11 and then the pH was changed by 9 after the uniform mixing of the medium for half an hour. The various germs occurred 50% and 100% at pH 8 and pH 7 of the non-sterilized medium, respectively. Production of oyster mushroom increased by $2,030\;ton\;ha^{-1}$ when the main raw material used corn pith instead of waste cotton. The time required of mycelium culture was 30 days when hypha was cultured at the non-sterilized medium, and pinhead occurred when 2 or 3 days was passed after the time required of mycelium culture. Occurrence of pinhead was most rapid at the condition of $22{\sim}26^{\circ}C$, 65% humidity and pH $6.5{\sim}7.0$ and required of $22{\sim}28$ days at $70{\sim}80\;mm$ thickness of non-sterilized medium. Ca content in 1st harvest oyster mushroom was higher than that in 2nd harvest one, and its difference was $30.3\;mg\;kg^{-1}$. Amino acid content by stipe thickness of oyster mushroom was ranged from 411.2 to $343.9\;mg\;kg^{-1}$ both in a pileus and a stipe of 1st harvest mushroom, and from 402.4 to $498.2\;mg\;kg^{-1}$ and from 442.6 to $470.4\;mg\;kg^{-1}$ in those of 2nd harvest one, respectively. The results of the present study suggest that the non-sterilization medium by addition of $Ca(OH)_2$ is usable with the cultivation of oyster mushroom.

In Vitro Studies on Pinus koraiensis (I) - Establishment and Growth of Callus - (잣나무의 기내배양(器內培養)에 관(關)한 연구(硏究) (I) - Callus 의 유발(誘發)과 생장(生長) -)

  • Park, Young Goo;Kim, Oue Ryong
    • Journal of Korean Society of Forest Science
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    • v.59 no.1
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    • pp.51-56
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    • 1983
  • Embryo tissue from Pinus koraiensis was established in culture on G.D. and W.S. basal medium supplemented with the auxin (NAA, 2,4-D, IBA) and kinetin. The various combinations of growth regulators were studied in order to determine the specific requirements of the callus tissue in vitro. The inorganic nutrient combination of G.D. medium was found to be better than that of W,S. medium. G,D. basal medium supplemented with 0.1 ppm NAA and 0.1 ppm kinetin was the most successful nutrient combination for the growth of callus induced from the embryo of P. koraiensis.

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Higher Biomass Production of Lactobacillus bulgaricus NLS-4 by Improvement of Cultural Conditions (배양조건 개선에 의한 Lactobacillus bulgaricus NLS-4의 균체 생산성 향상)

  • Rhee, Sang-Ki;Pack, Moo-Young
    • Microbiology and Biotechnology Letters
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    • v.8 no.1
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    • pp.55-60
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    • 1980
  • Some of the cultural conditions were improved in order to obtain the higher biomass of Lactobacillus bulgaricus NLS-4 which has the higher lactic acid producing activity as well. Among eight media including 11% non-fat milk medium as a control, the TIP medium was selected. By a batch experiment, the maximum cell concentration could be increased to 1.0$\times$10$^{9}$ cells per $m\ell$ when the organism was grown at 38$^{\circ}C$ for 18 hours with agitation speea of 200 rpm and under the constant level of pH 6.5 con-trolled with 1 N KOH solution in the selected medium. The cell concentration was further increased to 2.3$\times$10$^{9}$ cells per me in the steady state of continuous culture at the dilution rate of 0.17 hr$^{-1}$ for 18 hours.

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Improvement effect of total nitrogen and amino acid content in spent mushroom substrates by bacterial treatment (세균을 이용한 수확후배지의 총질소 및 아미노산 증진 효과)

  • Baek, Il-Sun;Kim, Jeong-Han;Lee, Yong-Seon;Shin, Bok-Eum;Lee, Young-Soon
    • Journal of Mushroom
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    • v.16 no.3
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    • pp.225-230
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    • 2018
  • This study was conducted to reuse spent mushroom substrates (SMS) of Pleurotus ostreatus and improve their nitrogen content by bacterial treatment. Two kinds of bacteria were used to investigate the increase in total nitrogen (T-N) content. Bacillus sp. (GM20-4) was isolated from SMS of oyster mushroom, and Rhodobacter sphaeroides (RS) was obtained from Gwangju Si Agricultural Technology Center. SMS samples were collected from three oyster mushroom cultivation farms located in Gyeonggi-do province, Korea. When dried SMS was inoculated with 30% culture broth of GM20-4 and RS and incubated at room temperature ($25{\pm}2^{\circ}C$) for 5 days, T-N content increased. To investigate the T-N content of other SMS, three dried SMS samples (A, B, and C) were treated by the same method using GM20-4 and RS. As a result, the T-N content of sample B was 20% higher than that of the control, whereas the T-N content of samples A and C increased to 17% and 12%, respectively. The change in T-N content by bacterial treatment of wet SMS was slightly higher than that of the control. The changes in amino acid content were also found to be higher than those in the control in all SMS samples by GM20-4 and RS treatment. Aspartic acid and glutamic acid contents were the highest among all amino acid compositions. Especially, the aspartic and glutamic acid contents of sample B increased by 2.9 folds higher than the control.

Effect of Growth Medium and Extract Sterilization on a Seedling Bioassay of Alfalfa Autotoxicity (생장배지 및 추출액의 멸균이 알팔파 Autotoxicity 의 유묘 생물검정에 미치는 영향)

  • Chon, Sang-Uk;Nelson, C.Jerry;Han, Sung-Uk;Guh, Ja-Ock
    • Korean Journal of Environmental Agriculture
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    • v.18 no.2
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    • pp.185-190
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    • 1999
  • Autotoxic substance(s) from alfalfa(Medicago sativa L.) plants reduces germination and growth of adjacent new alfalfa after alfalfa. The autotoxic chemical(s) in alfalfa are clearly unknown. Our objective was to improve the sensitivity of an alfalfa seedling bioassay for evaluating autotoxic leaf extracts. We determined critical extract concentrations that inhibit seed germination and seedling growth, compared two different culture media, and evaluated the effects of extract sterilization on the sensitivity of the assay, by using streptomycin and autoclaving method. An agar medium in petri plate gave better responses of germination and seedling growth to the extracts than using filter paper in the plate. On agar medium, the concentration of extract required to reach 50% inhibition of root length was 2.7 g $kg^{-1}$, and of germination and hypocotyl length were 3.8 and 9.9 g $kg^{-1}$, respectively. Leaf extracts with 100 ppm streptomycin stimulated germination significantly compared to Leaf extract alone but reduced root length of control by 43%. Root length was more sensitive to the autotoxin(s) than was germination or hypocotyl length. These results suggest that agar medium mixed with extract and sterilization by autoclaving could be improved the consistency and precision of bioassay, and that root length was the best parameter of autotoxic effect of alfalfa leaf extract.

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Multiple Shoot Induction and Bulb Mass Proliferation System by in Vitro Immature Spathe Culture of Elephant Garlic (Allium ampeloprasum L.) (코끼리마늘(Allium ampeloprasum L.)의 기내 미숙총포 배양을 통한 다신초유도와 종구대량증식 시스템)

  • Kwon, Young Hee;Jeong, Jae Hyun;Lee, Jae Sun;Jeon, Jong Ok;Park, Young Uk;Min, Ji Hyun;Chang, Who Bong;Lee, Sang Young;Youn, Cheol Ku;Kim, Ki Hyun
    • Korean Journal of Plant Resources
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    • v.31 no.4
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    • pp.355-362
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    • 2018
  • This study was performed to develop the mass propagation system using tissue culture technique to supply the seeds of Elephant garlic (Allium ampeloprasum L.) which has difficulty in propagation. Immature spathe of Elephant garlic was cultured on Murashige & Skoog (MS) medium supplemented with two plant growth regulators, naphthaleneacetic acid (NAA) and kinetin. After 6 weeks of culture, the highest number of shoot (14.9/explant) was obtained when the immature spathe with 10 cm length was cultured right after harvesting. In MS medium supplemented with 2 mg/L kinetin and 0.5 mg/L NAA, the most vigorous growth characteristics was observed, the shoot number was 14.9/explant, its length was 11.3 cm, and its fresh weight was 2.5 g. When the bulblets were cultured in MS medium with 2 mg/L kinetin and 0.5 mg/L NAA, the addition of 30 mg/L adenine improved their proliferation and growth significantly, the highest bulblet formation rate (48%) was obtained. The addition of 7% sucrose also increased the bulblet formation rate at the highest frequency of 98.2%. The shoots were shown be more vigorously proliferated at the secondary subculture stage rather than primary culture stage, their propagation rate was 80% after subculture.

Effects on yield of Pleurotus ostreatus(Suhanneutari 1ho) according to afterripening conditions with bottle cultivation (후숙 조건에 따른 느타리(수한느타리버섯 1호)의 생육특성)

  • Chang, Who-Bong;Kim, Min-Ja;Park, Jae-Seong;Choi, Jae-Sun;Song, In-Gyu;Jeong, Min-Jung;Cha, Jae-Soon
    • Journal of Mushroom
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    • v.10 no.2
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    • pp.63-67
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    • 2012
  • This study of afterripening conditions during oyster mushroom (Suhanneutari 1ho) cultivation in bottles was investigated. Medium materials were used poplar-sawdust (68%), cotton seed peel (16%), Beet pulp (8%) and cotton seed cake (8%). Mix of materials was used as a percentage of the volume and to adjust moisture content (67%). Autoclaved mediums were placed in low temperature storage ($20^{\circ}C$) and then moved in inoculation room and conducted mechanical inoculation. Mycelial culture temperature was maintained at $20{\sim}21^{\circ}C$ and cultured during 18 days. The afterripening period were 6days, 9days, 12days and 15 days at $25{\sim}27^{\circ}C$. The yield of fruit body was higher for 9 days (163.1g/bottle) and 12 days (160.7g/bottle) than that of other afterripening period. Second, in the changes in moisture content of the medium according to the afterripening period, no significant changes were observed during mycelial grwoth. The longer afterripening period was showed slightly lower weight of media. The moisture content of media after harvest at afterripening for 9 days had the biggest reduction than any other treatments. In addition, weight of media and yield of afterripening for 9 days were the lowest and highest, respectively.

Effects of Supplemental Levels of Spent Mushroom (Flammulina velutipes) Substrates on Chemical Composition and Quality of Whole Crop Sorghum Silage (팽이버섯 수확 후 배지의 첨가수준이 수수 사일리지의 화학적 조성과 품질에 미치는 영향)

  • Moon, Yea-Hwang;Lee, Sung-Sill;Kang, Tae-Won;Cho, Soo-Jeong
    • Journal of Mushroom
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    • v.10 no.3
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    • pp.136-142
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    • 2012
  • This study was carried out to determine the supplemental level of spent mushroom (Flammulina velutipes) substrates as an energy source in manufacturing of high moisture sorghum whole crop silage. Whole crop sorghum was harvested at heading stage and ensiled with spent mushroom substrates of 20% (S-20), 40% (S-40) and 60% (S-60) as fresh matter basis. Each silage was manufactured in plastic buckets included vinyl bag by three replications and stored for 0, 3, 6 and 9 weeks, respectively. Fermentation characteristics and quality of sorghum silages manufactured by supplemental level of spent mushroom substrates were as follows. Moisture contents of whole crop sorghum and spent mushroom substrates were 83.85% and 54.3%, respectively, and that of silages was 78% for S-20, 71% for S-40 and 68% for S-60. Ether extracts content of silages was significantly (P<0.05) increased during the fermentation periods. The pH in silages fermented for 3 weeks and above ranged from 4.24 to 4.42, and the decrease of pH by fermentation was relatively greater in S-40 compare to the other treatments. The lactic acid content of silage inclined that the S-40 was higher compared to the other treatments and decreased with elongation of fermentation period of silage. The contents of acetic acid and propionic acid of silages were not influenced by treatments and fermentation period. Flieg's score for estimation of silage quality ranged from 60 to 83, and was relatively high quality in the S-40 fermented for 9 week, and was relatively low quality in the S-60 fermented for 9 week. From above results, we suggest that 40% supplementation of spent Flammulina velutipes mushroom substrates as an energy source is resonable level in manufacturing of high moisture sorghum whole crop silage.