• Title/Summary/Keyword: Broth media

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Characteristics of Biosurfactant Producing Pseudomonas sp. HN37 (생물계면활성제 생성 세균 Pseudomonas sp. HN37의 특성)

  • Jung, Da Hee;Chang, Dong Ho;Kim, Yeong Eun;Jeong, Mi Rang;Hahn, Kyu Woong;Kim, Hyong Bai;Park, Kyeong Ryang
    • Korean Journal of Microbiology
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    • v.50 no.1
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    • pp.33-39
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    • 2014
  • One hundred forty four bacterial colonies which were able to degrade crude oil were isolated from soil samples that were contaminated with oil in Daejeon area. Among them, one bacterial strain was selected for this study based on its emulsification activity, growth rate and surface tension activity, and this selected bacterial strain was identified as Pseudomonas sp. HN37 through physiological- biochemical tests and analysis of its 16S rRNA sequence. Pseudomonas sp. HN37 utilize the several aliphatic hydrocarbons, 3,5-dinitrosalicylic acid and 2,4-dichlorophooxyacetic acid as a sole carbon source. And this bacterial strain showed a high resistance to antibiotics such as ampicillin and chloramphenicol, as well as heavy metals such as Ba, Cr, Li and Mn. Also, it was found that the optimal pH and temperature for the cell growth, surface tension, and emulsification activity of Pseudomonas sp. HN37 were pH 6.0-9.0 and $30^{\circ}C$, respectively. The emulsification and surface tension activity was reached the maximum to 1% (V/V) crude oil and 1% (W/V) NaCl concentration. The surface tension of the culture broth was decreased from 62 to 27 dyne/cm after fifteen hours of inoculation in LB media.

Studies on the EPA(eicosapentaenioc acid) production by marine bacteria (해양세균(海洋細菌)을 이용(利用)한 EPA(eicosapentaenoic acid) 생성(生成)에 관한 연구(硏究))

  • Joh, Yong-Goe;Kim, Seong-Jin;Kim, Ji-Soo;Lee, Min-Kyeng
    • Journal of the Korean Applied Science and Technology
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    • v.12 no.2
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    • pp.121-130
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    • 1995
  • This project has been worked out for isolation of EPA-producing bacteria from marine source of sea water, sea sediment and intestinal contents eviscerated from some red-muscle fish such as mackerel, horse-mackerel and spike fish. The samples were precultured on the media of PPES-II glucose broth and then pure-cultured on Nutrient agar and P-Y-M glucose. Lipids extracted from those bacterial mass collected by centrifugation were analysed in terms of lipid class and fatty acid composition. The results are resumed as follows : 1. 112 strains from sea water and 76 strains from sea sediment were tested for their EPA producing capability, but both strains of (SA-67 and SA-91) from the former and four strains(SS-35, 37, 51 and 71) from the latter have been proved to produce EPA above the level of 2% of total fatty acids. The strains such as GS-11, 29, 31, HM-9, 29, B-18, 33, 107, YL-129, 156, 203, 77, 104 and 256 which were isolated from fish intestinal contents, have also produced EPA at higher level than 2% of total fatty acids. 2. Contents of total lipids extracted from the cultures of these strains grown at $25^{\circ}C$, range from 2.8% to 6.9% (on dry weight %), and they are mainly composed of polar lipids($40.9{\sim}52.9%$) such as phosphatidyl glycerol($^{+}cardiolipin$)(?) and phosphatidyl ethanolamine ($33.8{\sim}40.0%$), with smaller amount of free fatty acid ($11.2{\sim}20.2%$). 3. EPA was isolated from a mixture of fatty acid methyl esters obtained from the lipid of each strain by HPLC in silver-ion mode and was identified by GC-Mass spectrometry. 4. The strains of SW-91, GS-11, GS-29, HM-9, B-18 and YL-203 grown at $25^{\circ}C$ have a level of 5% EPA in their total fatty acids, and the GS-11 and HM-9 strains show a tendency of increase in the EPA level with an increase of growth temperature.

The Frequency of Detecting Prevotella intermedia and Prevotella nigrescens in Korean Adult Periodontitis Patients (한국인 치주 감염 환자에서의 Prevotella intermedia와 Prevotella nigrescens의 발현빈도)

  • Peck, Seung-Yup;Ku, Young;Rhyu, In-Cheol;Hahm, Byung-Do;Han, Soo-Boo;Choi, Sang-Mook;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.30 no.2
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    • pp.419-429
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    • 2000
  • Prevotella intermedia has been implicated as a potent pathogen in many kinds of periodontal, pulpal and periapical diseases. However, it has been isolated from periodontally healthy adults and from edentulous children as well. The intraspecies heterogeneity of Prevotella intermedia has been demonstrated in early studies and finally Shah & Gharbia confirmed the existence of 2 DNA homology groups and proposed dividing Prevotella intermedia into 2 species, Prevotella intermedia and Prevotella nigrescens. This study was designed to examine the frequency of Prevotella intermedia and Prevotella nigrescens in diseased periodontal pockets and healthy gingival sulcus of Korean people by PCR based on 16s ribosomal DNA sequence. One hundred adults who had adult periodontitis but not taken any periodontal treatment or antibiotics during previous 6 months and 50 adults who had healthy periodontal tissue were selected for this study. The sulcular fluid was collected into VMGA by sterilized paper point and diluted to 1,000 times in anaerobic chamber. $100{\mu}{\ell}$ of sample was cultured in $37^{\circ}C$ for 10 days. Among the bacterial colonies, BPB were selected and cultured in BHI broth and then Prevotella intermedia was identified through Gram staining and biochemical test. Identified Prevotella intermedia was cultured again and centrifuged. DNA was extracted from the pellet using several reagents. PCR was performed by previously designed primer. The results were followed. 1. BPB were isolated from 39 of 100 samples of diseased periodontal pockets(39%). 2. Prevotella intermedia was identified from 24 of 39 BPB samples. 3. Among 24 Prevotella intermedia, 21 were confirmed as Prevotella inter - media(87.5) and 2 were confirmed as Prevotella nigrescens(8.33%). 4. BPB were isolated from 9 of 50 samples of periodontally healthy patients. Among them only two were identified as Prevotella intermedia, that is, one was confirmed as Prevotella intermedia and the other was Prevotella nigrescens.

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Studies on Cultural Characteristics Pestalotiopsis theae causing Leaf Blight on Oriental Persimmon Tree (단감나무 둥근갈색무뉘병원균 Pestalotiopsis theae의 배양적 특성)

  • Chang, Tae-Hyun;Lim, Tae-Heon;Chung, Bong-Koo;Kim, Byung-Sup
    • Korean Journal Plant Pathology
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    • v.13 no.4
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    • pp.232-238
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    • 1997
  • Culture conditions affecting mycelial growth and sporulation of P. theae, SP2, SP3 and P. longiseta which causing leaf blight on oriental persimmon leaf blight have been investigated. The optimum temperature for the mycelial growth and sporulation on potato dextrose agar was $25{\sim}30^{\circ}C$ in all the fungi, but was inhibited and finally arrested at 10 and $30^{\circ}C$. The optimum pH for mycelial growth and sporulation were ranged at pH 4.5~5.0 and 5.0~6.0 in all the fungi. Lenonian agar, potato sucrose agar and oatmeal medium were good culture media for the mycelial growth and sporulation of all the fungi. The effective sources of nitrogen and carbon for the mycelial growth were tryptone, glycine, starch, dextrose, galactose and lactose. Glutamic acid, peptone and tryptone were good nitrogen sources for sporulation of the fungi. Sucrose, starch and galactose were also good carbon sources for sporulation. Generally, vitamins had no effect on mycelial growth and sporulation. The pH of the potato dextrose broth inoculated with SP2, SP3 and P. theae and P. longiseta was changed from 7.0 to 4.5~4.7 and 5.0~5.4 after incubating for 10 days, respectively. But, the initial pH of the medium adjusted to 5.0 was lowered to 4.5~4.7 after incubating for 10 days.

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Selection and Enumeration of Bifidobacteria in Fermented Foods with Various Lactic Bacteria (다양한 유산균이 존재하는 발효식품으로 부터 Bifidobacteria의 선별 및 계수)

  • Lee, Si-Kyung;Park, Dong-Ki;Oh, Hoon-Il;Park, Jong-Hyun
    • Korean Journal of Food Science and Technology
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    • v.31 no.3
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    • pp.751-756
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    • 1999
  • The inhibitory effect of antibiotics on growth of lactobacillus, streptococcus and bifidobacteria was examined to develop the selective media to isolate and enumerate bifidobacteria from the fermented foods containing various lactic bacteria. The growth of lactic bacteria was inhibited seriously but that of bifidobacteria was not inhibited by gentamycin or ripampicin at the concentration of more than $100\;{\mu}g/mL$. However lactic bacteria did not grow in MRS broth containing $50\;{\mu}g/mL$ of ampicillin and the growth inhibition of bifidobacteria occurred. The growth inhibition of bifidobacteria was more severe than lactic bacteria in $100\;{\mu}g/mL$ of fosfomycin. Therefore, the MRS medium containing $80\;{\mu}g/mL$ of neomycin sulfate, $50\;{\mu}g/mL$ of gentamycin, $50\;{\mu}g/mL$ of rifampicin, $15\;{\mu}g/mL$ of nalidixic acid and $3\;{\mu}g/mL$ of lithium chloride was concluded selective for bifidobacteria, but restrictive for the other lactic bacteria present in Kimchi and cheese.

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Combination Effects of Potassium Sorbate and Sodium Benzoate with sodium Chloride on the Growth Inhibition of Escherichia coli and Salmonella typhimurium (Escherichia coli 와 Salmonella typhimurium 의 생육억제에 미치는 식염과 Potassium Sorbate, Sodium Benzoate의 병용효과)

  • Cho, Nam-Sook;Yang, Yeo-Young;Choi, Eon-Ho
    • Korean Journal of Food Science and Technology
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    • v.18 no.4
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    • pp.249-254
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    • 1986
  • An experiment was performed to investigate the combined effect of preservatives and the synergistic effect of sodium chloride to them on the inhibition of bacterial growth. Escherichia coli and Salmonella typhimurium were cultured with or without shaking in liquid media (pH 6) of tryptone-glucose-yeast extract or tryptic soy broth which contained 0.1% potassium sorbate and/or 0.03% sodium benzoate, equivalent to half of the maximum permissible levels, respectively. The growth of E. coli was more inhibited with one or both of the two preservatives by shaking culture than by non-shaking culture. For S. typhimurium the single treatment of the preservatives did not show inhibitory effect whereas the combined treatment of them showed bacteriostatic effect in shaking culture and a prolongation of lag phase in non-shaking culture. Addition of 2% sodium chloride to either potassium sorbate or potassium sorbate plus sodium benzoate remarkably increased the growth inhibition of E. coli for non-shaking cultivation but no effect observed for shaking cultivation. S. typhimurium was more sensitive to the addition of sodium chloride than E. coli in both shaking and non-shaking culture to show lower viable cell counts than initial numbers.

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Preparation of $Gal^3Man_4(6^3-mono-{\alpha}-D-galacto-pyranosyl-{\beta}-mannotetraose)$ by Bacillus sp. ${\beta}-mannanase$ and Growth Activity to Intestinal Bacteria (Bacillus sp.유래 ${\beta}-mannanase$에 의한 $Gal^3Man_4(6^3-mono-{\alpha}-D-galacto-pyranosyl-{\beta}-mannotetraose)$ 조제 및 장내세균에 대한 생육활성)

  • Kim, Sang-Woo;Park, Gwi-Gun
    • Applied Biological Chemistry
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    • v.47 no.4
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    • pp.379-383
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    • 2004
  • For the elucidation of substrate specificity to the brown copra meal by Bacillus sp. ${\beta}-mannanase.$, the enzymatic hydrolysate after 24 hr of reaction was heated in a boiling water bath for 10 min, and then centrifuged to remove the insoluble materials from hydrolysates. The major hydrolysates composed of D.P 5 and 7 galactosyl mannooligosaccharides. For the separate of galactosyl mannooligosaccharides, the supernatant solution of 150 ml was put on a first activated carbon column. The column was then washed with 5 l of water to remove mannose and salts. The oligosaccharides in the column were eluted by a liner gradient of $0{\sim}30%$ ethanol, at the flow rate of 250 ml per hour. The sugar composition in each fraction tubes was examined by TLC and FACE analysis. The combined fraction from F3 was concentrated to 30 ml by vacuum evaporator. Then put on a second activated carbon column. The oligosaccharides in the column were eluted by a liner gradient of $0{\sim}30%$ ethanol (total volume: 5 l), at the flow rate of 250 ml per hour. The eluent was collected in 8 ml fraction tubes, and the total sugar concentration was measured by method of phenol-sulfuric acid. The major component of F2 separated by 2nd activated carbon column chromatography were identified $Gal^3Man_4(6^3-mono-{\alpha}-D-galactopyranosyl-{\beta}-mannotetraose)$. To investigate the effects of brown copra meal galactomannooligosaccharides on growth of Bifidobacterium longum, B. bifidum were cultivated individually on the modified-MRS medium containing carbon source such as $Gal^3Man_4$, compared to those of standard MRS medium.

Characteristics of Soycurd-forming Lactic Acid Bacteria that Produce Gammaaminobutyric Acid (GABA) from Kimchi (김치로부터 GABA를 생산하는 커드 형성 젖산균의 특성)

  • Kim, Eun-Ah;Mann, So-Yon;Kim, Su-In;Lee, Ga-Young;Lee, Byong-Won;Kim, Dong-Seob
    • Journal of Life Science
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    • v.24 no.1
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    • pp.46-53
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    • 2014
  • Lactobacillus sakei 383, which showed the highest GABA content in fermented soycurd, survived in artificial gastric fluid (pH 3.0) up to 3 h, and the survival rate was 88%. L. sakei 383 was tolerant to bile juice during incubation in MRS broth with 0.3% oxgall, and the survival rate was 99%. The survival ratio of L. sakei 383 was high in media containing less than 6% NaCl. L. sakei 383 produced an antibacterial substance against various pathogens, including Listeria monocytogenes, Bacillus cereus, Staphylococcus aureus, Escherichia coli, and Salmonella typhi. The quality characteristics of soycurd fermented with L. sakei 383 were measured during the fermentation period. The viable cell number reached a peak ($10^{11}CFU/ml$) 36 h after fermentation and then slowly decreased. According to the fermentation time of L. sakei 383, the acidity of soycurd increased and the pH decreased until 12 h, and they were maintained thereafter. The moisture, crude ash, crude protein, crude fat, and crude fiber content was 94.88, 0.22, 2.38, 1.16, and 0.03%, respectively. The content of total and reducing sugar was comparatively higher in the soycurd fermented with L. sakei 383 than in nonfermented soycurd. The essential and nonessential amino acid content was 11.2 and 38.65 mg/100 g.

The Optimal Culture Conditions and Antifungal Activity of Culture Extract from Oudemansiella mucida (끈적긴뿌리버섯(Oudemansiella mucida)의 최적배양조건 및 배양 추출액의 항균작용에 관한 연구)

  • Choi, Mi-Ryue;Cho, Hae-Jin;Lee, Jae-Seong;Kim, Hye-Young;Lee, Tae-Soo
    • The Korean Journal of Mycology
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    • v.39 no.2
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    • pp.91-98
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    • 2011
  • Oudemansiella mucida, an edible and medicinal mushrooms belonging to Tricholomataceae of Basidiomycota, has been known to produce antifungal substances to inhibit the mycelial growth and spore germination of the plant pathogenic fungi. To produce good amount of antifungal substances from culture media, the optimal culture conditions of O. mucida were investigated. The most favorable conditions for the mycelial growth were $25^{\circ}C$ and pH 5 in potato dextrose agar. The most favorable carbon and nitrogen sources promoting mycelial growth were maltose and calcium nitrate, respectively. The optimum C/N ratio was about 20 : 1 in case that 3% glucose was supplemented to the basal medium as a carbon source. The optimal mycelial growth of O. mucida was found in the Hennerberg medium. The crude extract from submerged culture of potato dextrose broth exhibited inhibition of mycelial growth of Colletotrichum acutatum, Botrytis cinerea and Pyricularia oryzae but, fungicidal activity is not good enough to compared with commercially available fungicides tested. Therefore, the antifungal substances extracted from submerged culture of O. mucida might have a potential to be used for biocontrol agent of fungal diseases of plants.

Biological Control of Lettuce Sclerotinia Rot by Bacillus subtilis GG95 (길항미생물 Bacillus subtilis GG95를 이용한 상추 균핵병의 생물학적 방제)

  • Lee, Hyun-Ju;Kim, Jin-Young;Lee, Jin-Gu;Hong, Soon-Sung
    • The Korean Journal of Mycology
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    • v.42 no.3
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    • pp.225-230
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    • 2014
  • Sclerotinia sclerotiorum, a plant pathogenic fungus, can cause serious yield and quality losses in the winter lettuce field. For biological control of S. sclerotiorum, soil-born microorganisms that inhibit the mycelia growth of S. sclerotiorum and Fusarium oxysporum were isolated from diseased soil. Among the isolates, bacterial isolate, GG95, which was identified as Bacillus subtilis according to the morphological, physiological characteristics and by 16S rRNA similarity, showed the highest level of inhibitory activity. The growth conditions for B. subtilis GG95 were optimized in TSB media (pH 7) by culturing at $28^{\circ}C$ for 24 hrs. Maltose or fructose and peptone were selected as the best carbon and nitrogen sources, respectively. Greenhouse experiment was performed to test effectiveness of B. subtilis GG95 in the control sclerotinia rot. Drench application ($1{\times}10^8cfu/mL$, 3 times) of the bacterial culture broth to lettuce showed an effectiveness value of 88%, suggesting that B. subtilis GG95 would be a promising biocontrol agent for control of sclerotinia rot.