• Title/Summary/Keyword: culture broth

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Production and Characterization of Extracellular Phospholipase D from Streptomyces sp. YU100

  • Lim, Si-Kyu;Choi, Jae-Woong;Chung, Min-Ho;Lee, Eun-Tae;Khang, Yong-Ho;Kim, Sang-Dal;Nam, Doo-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.12 no.2
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    • pp.189-195
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    • 2002
  • Using Streptomyces sp. YU100 isolated from Korean soil, the fermentative production of phospholipase D was attempted along with its purification and characterization studies. When different carbon and nitrogen sources were supplemented in the culture medium, glucose and yeast extract were found to be the best. By varying the concentration of nutrients and calcium carbonate, the optimal culture medium was determined as 2.0% glucose, 1.5% yeast extract, 0.5% tryptone 0.3% calcium carbonate. During cultivation, the strain secreted most of the phospholipase D in the early stage of growth within 24 h. The phospholipase D produced in the culture broth exhibited hydrolytic activity as well as transphosphatidylation activity on lecithin (phosphatidylcholine). In particular, the culture broth showed 8.7 units/ml of hydrolytic activity when cultivated at $28^{\circ}C$ for 1.5 days. The phospholipase D was purified using 80% ammonium sulfate precipitation and DEAE-Sepharose CL-6B column chromatography, which produced a major band of 57 kDa on a 10% SDS-polyacrylamide gel with purity higher than 80%. The enzyme showed an optimal pH of 7 in hydrolytic reaction, and at pH 4 in a transphosphatidylation reaction. The enzyme activity increased until the reaction temperature was elevated to $60^{\circ}C$. The enzyme was relatively stable at high temperatures and neutral pH, but significantly unstable in the alkaline range. Among the detergents tested as emulsifiers of phospholipids, the highest enzyme activity was observed when 1.5% Triton X-100 was employed. However, no inhibitory effect by metal ions was detected. Under optimized reaction conditions, the purified enzyme not only completely decomposed PC to phosphatidic acid within 1 h, but also exhibited higher than 80% conversion rate of PC to PS by transphosphatidylation within 4 h.

Measures to Improve Culinary Trends in Korean Court Food Based on the Perception of Korean Royal Court Cuisine (궁중음식 인식성향에 따른 궁중음식 메뉴개발 방향성에 대한 조사)

  • Koo, Ha Yeon;Choung, Seo Yeong;Jeong, Hee Sun
    • Korean journal of food and cookery science
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    • v.32 no.3
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    • pp.370-381
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    • 2016
  • Purpose: This study assesses Korean royal court cuisine as perceived by culinary professionals and students for the development of Korean dining. Methods: It was observed in a survey that Korean royal cuisine could be grouped into four classes represented by the following factors: popularity, standardization, tradition, and haute-cuisine. Results: From the analysis of the survey results, it was determined that the people surveyed could be grouped into the following three categories: those who prefer standardization/pursuit of haute-cuisine, traditionalists/popularizers, and those who are indifferent. The survey also assessed whether the ten most popular Korean dishes served to foreigners had variations in royal court cuisine and which food ingredients and combinations of dishes would be the most appropriate. It was determined that control over the sweetness when cooking Bulgogi was needed. For food usually consumed for invigoration, especially for the broth of soup dishes in summer, women preferred clear meat broth with soup than men. When preparing Japchae, it was found that control over the ratio between glass noodles and vegetables and control over the sweetness were needed with respect to the main dishes. Conclusion: The indicator 'education on Korean royal court cuisine culture' showed relatively low satisfaction compared to its high importance, implying that further improvement in these development measures is especially required.

Microbial Conversion of Ginsenoside $Rb_1$ to Minor Ginsenoside $F_2$ and Gypenoside XVII by Intrasporangium sp. GS603 Isolated from Soil

  • Cheng, Le-Qin;Na, Ju-Ryun;Kim, Myung-Kyum;Bang, Myun-Ho;Yang, Deok-Chun
    • Journal of Microbiology and Biotechnology
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    • v.17 no.12
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    • pp.1937-1943
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    • 2007
  • A new strain, GS603, having ${\beta}$-glucosidase activity was isolated from soil of a ginseng field, and its ability to convert major ginsenoside $Rb_1$ to minor ginsenoside or gypenoside was studied. Strain GS603 was identified as an Intrasporangium species by phylogenetic analysis and showed high ginsenoside-converting activity in LB and TSA broth but not in nutrient broth. The culture broth of the strain GS603 could convert ginsenoside $Rb_1$i into two metabolites, which were analyzed by TLC and HPLC and shown to be the minor ginsenoside $F_2$ and gypenoside XVII by NMR.

Isolation and Characterization of Bacillus cereus A-139 Producing Auxin from East Coast Sand Dunes (동해안 사구로부터 Auxin을 생산하는 Bacillus cereus A-139의 분리 및 그 특성)

  • So, Jai-Hyun;Kim, Duk-Jin;Shin, Jae-Ho;Yu, Choon-Bal;Rhee, In-Koo
    • Korean Journal of Environmental Agriculture
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    • v.28 no.4
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    • pp.447-452
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    • 2009
  • A bacterium, which was named to be Bacillus cereus A-139, secreting auxin was isolated from the east coast sand dunes in Korea. The secretion of auxin was confirmed by HPLC. When cultured in LB broth, Bacillus cereus A-139 produced $16.12\;{\mu}$g/mL auxin after 8 days in LB broth. Bacillus cereus A-139 produced $49\;{\mu}$g/mL auxin and $162.6\;{\mu}$g/mL by the addition of 2% tryptone and 0.1% tryptophan, respectively. The root growth of Arabidopsis thaliana was retarded by Bacillus cereus A-139 culture broth up to 57% but the formation of lateral roots was increased up to almost twice after 4 days incubation. Also the formation of lateral roots of mung bean was increased up to 57% after 10 days incubation.

Zinc Metal Solubilization by Gluconacetobacter diazotrophicus and Induction of Pleomorphic Cells

  • Saravanan, Venkatakrishnan Sivaraj;Osborne, Jabez;Madhaiyan, Munusamy;Mathew, Lazar;Chung, Jong-Bae;Ahn, Ki-Sup;Sa, Tong-Min
    • Journal of Microbiology and Biotechnology
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    • v.17 no.9
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    • pp.1477-1482
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    • 2007
  • Gluconacetobacter diazotrophicus strain PA15 exhibited a minimum inhibitory concentration value of 11 mM in an LGI medium amended with $ZnCl_2$. When an LGI medium was amended with Zn metal, solubilization halos were observed in a plate assay, and further solubilization was confirmed in a broth assay. The maximum solubilization was recorded after 120 h with a 0.1% Zn metal amendment. During solubilization, the culture growth and pH of the broth were indirectly correlated. Using a Fourier Transform Infrared Spectroscopy analysis, one of the agents solubilizing the Zn metal was identified as gluconic acid. When the Zn-amended broth was observed under a bright field microscope, long involution cells were observed, and further analysis with Atomic Force Microscopy revealed highly deformed, pleomorphic, aggregate-like cells.

Studies on the Conditions of Enzyme Production of Endocellular Cytosine Deaminase from Aspergillus fumigatus IFO 5840 (Aspergillus fumigatus IFO 5840의 균체내 Cytosine Deaminase의 생성에 관한 연구)

  • 김재근;하영득
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.20 no.2
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    • pp.179-186
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    • 1991
  • The nutritional requirement and cultural condition such as carbon and nitrogen sources, cultural temperature, initial pH, cultural time and aeration for the production of endocellular cytosine deaminase from Aspergillus fumigatus IFO 5840 were investigated. The cultural broth giving maximum cytosine deaminase yield was found to consist of 2% glucose as a carbon source and 1% yeast extract and 0.1% peptone as a nitrogen source. Optimal initial pH of the culture broth was pH 8.5 and the enzyme production in the cell usually reached a maximum after 28 hours of cultivation in the 500ml shaking flask containing 100ml broth at $30^{\circ}C$. The endoenzyme production of the used strain was inhibited by inorganic nitrogen, but activated by orgainc nitrogen, yeast extract.

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THE INHIBITORY EFFECT OF STREPTOCOCCUS SALIVARIUS 119 ON THE FORMATION OF ARTIFICIAL PLAQUE (Streptococcus salivarius 119의 인공치태 억제효과에 대한 연구)

  • Lee, Min-Ha;Yang, Kyu-Ho;Oh, Jong-Suk
    • Journal of the korean academy of Pediatric Dentistry
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    • v.27 no.1
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    • pp.15-23
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    • 2000
  • Streptococcus salivarius is a normal inhabitant in the human oral cavity. Streptococcus salivarius 119 in this study was isolated from the oral cavity of child and identified, and its action mechanism on the formation of denal plaque by Streptococcus matans was studied. 1. The optical density of absorbance at 550 nm was 0.327 in the culture of Streptococcus mutans in disposable cuvette, whereas being 0.119 in the combined culture of Streptococcus mutans and Streptococcus salivarius 119. 2. The mean weight of produced artificial plaque on the wires in the beaker was 84.7mg in culture of Streptococcus mutans only, whereas being reduced to 12.3mg in the combined culture of Streptococcus mutans and Streptococcus salivarius 119. 3. When Streptococcus mutans was cultured in the media containing culture supernatant of Streptococcus salivarius 119 in BHI broth, the mean weight of produced artificial plaque was 100.5mg on the wires, whereas being reduced to 20.4mg in the media containing culture supernatant of Streptococcus salivarius 119 in BHIS broth. 4. The viable cells of Streptococcus oralis and Streptococcus salivarius 119 were $4.8\times10^7\;and\;7.5\times10^8$ per ml respectively after each culture, wheras being $4.2\times10^7\;and\;5.8\times10^7$ per ml in the combined culture of Streptococcus oralis and Streptococcus salivarius 119. 5. The polymer produced by Streptococcus salivarius 119 was glucan on the thin layer chromatography. 6. The glucan produced by Streptococcus salivarius 119 was water-soluble glucan containing $1\rightarrow6$ linkages as the main linkage on the thin layer chromatography. These results suggested that isolated Streptococcus salivarius 119 inhibited the formation of artificial plaque by the production of water-soluble glucan.

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Development of an in vitro culture method for harvesting the free-living infective larvae of Strongyloides venezuelensis (베네수엘라분선충 (Strongyloides venezuelensis Brumpt, 1934) 자유생활형 유충의 시험관 내 배양 기술 개발)

  • ;M.
    • Parasites, Hosts and Diseases
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    • v.36 no.1
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    • pp.15-22
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    • 1998
  • An in uitro culture technique was established for harvesting Strongwloides venezuelensis free-living infective larvae using a nutrient broth medium as a substitute for rat-feces in polyvinyl culture bags ($10{\;}{\times}{\;}12{\;}cm$). The egg hatch rate V) in sterile saline at different incubation temperatures (X) was expressed as the quadratic function, Y = $-0.192X^2$ + 8.673x - 19.550 (r = 0.901). The highest (100%) egghatch rate was observed at $25^{\circ}C$. A significant difference (p<0.05) in development rate W) of free-living infective larvae was observed between different concentrations of nutrient broth (X) which was highest (20.6%1 in 0.12% nutrient broth concentrations, incubated at $20^{\circ}C$ for 5 days [Y = $-864.032X^2$ + 245.995X- 0.560 (r = 0.875)]. Yields (Y) of infective larvae were observed relatively high when the culture medium was incubated at higher temperatures (X) which peaked at $25^{\circ}C$ (20.0%) than at lower temperatures. $15^{\circ}C$M (10.9%) and $20^{\circ}C$ (18.1%) [Y = $-0.189X^2$ + 8.387x- 72.795 (r = 0.981)]. The period W) required for the development of infective larvae decreased with higher incubation temperatures (X) [Y = $0.035X^2$ - 2.025X + 32.375 (r = 0.995)] The highest yield (19.2%) of infective larvae was obtained from culture bag inoculated with 15.000 eggs than with below and over 15,000 eggs in 0.12% nutrient broth and incubated at $25^{\circ}C$ for 4 days. The newly adapted culture method (from egg to third-stage larva) may be useful as a bio-bar/bioassay system for screening new chemical products, anthelmintics and pesticides, as well as for parasito immunological studies with Strongwloides species.

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Effect of Culture Medium Containing Chicken Serum on Growth and Differentiation of QM7 Quail Muscle Cells (닭 혈청을 포함한 배양액 조성 변화가 QM7 메추리 근육세포의 성장 및 분화에 미치는 영향 분석)

  • Choi, Sarang;Lee, Sang In;Shin, Sangsu
    • Korean Journal of Poultry Science
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    • v.49 no.2
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    • pp.109-114
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    • 2022
  • QM7 cell, a quail muscle cell line, has been used in various studies. In cell culture, it is well known that the culture medium has a significant influence on cell growth and maintenance of cell characteristics. This study aimed to adjust the culture medium to make it more suitable for QM7 muscle cells. A culture medium was prepared by adding 2% chicken serum (CS) instead of 10% tryptose phosphate broth (TPB) to the conventional culture medium. In the culture medium prepared with CS, the QM7 muscle cells changed from a pointed, thin to a broader shape. In addition, they grew and divided more rapidly in the new culture medium than in the conventional culture medium. The number of cells increased faster in the CS-containing culture medium from day two after passaging, and significantly increased from day three. The muscle cells grown in the medium containing CS maintained their undifferentiated state prior to differentiation. Myotubes formed well when the cells were maintained in CS-containing medium, resulting in a longer length and uniformity. According to the above results, it is clear that the culture of QM7 myocytes using a medium containing CS rather than TPB helps obtain better results in cell maintenance and differentiation.

A simple culture technique of Rhodobacter azotoformans EBN-7 for public use: application to NH4+-N removal in shrimp aquaculture water

  • Cho, Kyoung Sook;Kim, Joong Kyun
    • Fisheries and Aquatic Sciences
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    • v.25 no.10
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    • pp.525-536
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    • 2022
  • Photosynthetic bacteria (PSB) attract considerable interest as useful microorganisms; nevertheless, a generalized culture technique has not been previously reported owing to difficulty in their cultivation. Therefore, a simple culture technique suitable for public use was investigated. Among the PSB tested, the strain Rhodobacter azotoformans EBN-7 was the most suitable for scale-up production because it showed the highest specific growth rate (0.20 h-1) on basal medium. In scale-up cultivation (500 L), R. azotoformans EBN-7 showed 4.50 × 1010 colony-forming units mL-1 (number of viable cells), dry cell weight of 26.8 g/L, and a specific growth rate of 0.15 h-1. Cultivation using this final culture broth (as seed culture) in a 15 L simple reactor was successful, with maintenance of cell activity evident. For use as seed culture, the maximum allowable preservation period of R. azotoformans EBN-7 at 4℃ was 3 months. When R. azotoformans EBN-7 cultivated in a simple technique was applied to shrimp aquaculture water, NH4+-N was reduced from 0.61 mg/L to 0.24 mg/L (by 60.7%) in 4 days in comparison with the control. Thus, this simple culture technique using R. azotoformans EBN-7 has the potential for a good removal efficiency of NH4+-N, making seed culture easier and suitable for public use.