• Title/Summary/Keyword: shake flask

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Non-invasive Methods for Determination of Cellular Growth in Podophyllum hexandrum Suspension Cultures

  • Chattopadhyay, Saurabh;Bisaria, V.S.;Scheper, T.;Srivastava, A.K.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.6
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    • pp.331-334
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    • 2002
  • Culture conductivity and on-line NADH fluorescence were used to measure cellular growth in plant cell suspension cultures of Podophyllum hexandrum. An inverse correlation between dry cell weight and medium conductivity was observed during shake flask cultivation. A linear relationship between dry cell weight and culture NADH fluorescence was obtained during the exponential phase of batch cultivation In a bioreactor under the pH stat (pH 6) conditions. It was observed that conductivity measurement were suitable for biomass characterisation under highly dynamic uncontrolled shake flask cultivation conditions. However, if the acid/alkali feeding is done for pH control the conductivity measurement could not be applied. On the other hand the NADH fluorescence measurement allowed online-in situ biomass monitoring of rather heterogenous plant cell suspension cultures in bioreactor even under the most desirable pH stat conditions.

A Study on the Antibacterial Activity of Chitosan

  • Chli, In-Ryu;Park, Jeong-Im
    • Proceedings of the Costume Culture Conference
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    • 2003.02a
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    • pp.24-25
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    • 2003
  • Water-soluble chitosan and water-insoluble chitosan with molecular weight of 2,000,000, 500,000, 80,000, and 40,000 and more than 90% degree of deacetylation were controlled to evaluate the antibacterial activity of chitosan against a pathogenic bacteria, methicillin resistant Staphylococcus aureus(MRSA), which is being issued in the world. The Shake Flask Method and Modified Shake Flask Method were used to find out the antibacterial activities of 5types of chitosan/acetic acid solution, and the other antibacterial activities test with the cotton filter treated with chitosan /acetic acid solution. Those test methods showed the great differences ,but the results of the antibacterial activites showed the same difference.

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Characterization of the nar Promoter of Escherichia coli to use as an inducible promoter in Wild-type host Agrobacterium.tumefaciens

  • Lee, Gil-Ho;Jo, Mu-Hwan;Lee, Jong-Won
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.758-761
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    • 2001
  • In this study, the nar promoter of E. coli was characterized to see whether the nar promoter cloned onto pBBR122 can be used as an expression promoter of gram negative microbes. For this purpose, a plasmid with lacZ gene expressing ${\beta}-galactosidase$ instead of the structural genes of nar operon in a gram negative host strain(Agrobacterium.tumefaciens) was used to simplify an assay of induction of the nar promoter. The following effects were investigated to find optimal conditions: methods of inducing the nar promoter, optimal nitrate concentration, maximally inducing the nar promoter, the amount of expressed ${\beta}-galactosidase$ and induction ratio(specific ${\beta}-galactosidase$ activity after maximal induction/specific ${\beta}-galactosidase$ activity before induction). The following results were obtained from the experiments: the growth of Agrobacterium with E.coli nar promoter was not much affected by nitrate concentration in the shake-flask; induction of nar promoter was optimal when Agrobacterium was grown in the presence of 1% nitrate ion at the beginning of culture and when overnight culture was completely grown in the shake-flask before being transferred to other shake-flask; the amount of ${\beta}-galactosidase$ per cell and per medium volume was maximal when Agrobacterium was grown under aerobic condition to $OD_{600}$ of 1.7; then the nar promoter was induced under microaerobic and anaerobic condition made by lowering dissolved oxygen level(DO). After 2-3h of induction in the YEP medium selected as a main culture medium, the specific ${\beta}-galactosidase$ activity became about 17,000 Miller units in the fermentor cluture.

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Growth Condition of Liquid Culture by Pleurotus ostreatus (액체배양에서 느타리버섯균의 적합한 생장조건 구명)

  • Sung, Jae-Mo;Moon, Hee-Woo;Park, Dong-Soo
    • The Korean Journal of Mycology
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    • v.27 no.1 s.88
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    • pp.1-9
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    • 1999
  • For the practical using of liquid spawn we carried out selection test of nutrient sources, cultural methods and cultural apparatus for liquid spawn production of oyster mushroom(ASTI 2001, ASTI 2018, ASTI 2072, ASTI 2016, ASTI 2070, ASTI 2180, ASTI 2183, ASTI 2042). The optimal temperature and pH range for mycelial growth of Pleurotus species were $25^{\circ}C\;to\;30^{\circ}C$ and 5.5 to 6.5, respectively. The effect of carbon sources, nitrogen sources and mineral salts on the mycelial growth was studied using petridish culture. Generally, the disaccharides and polysaccharides showed good effect for mycelial growth of Pleurotus species, and the polysaccharides were superior to the other classes of carbon sources for mycelial growth. For the mycelial growth of the 8 oyster mushroom stains, soybean flour was superior to the other kinds of nitrogen sources. On the other hand, addition of mineral salts did not affect, and even poor under certain mineral salts, the mycelial growth of the 8 oyster mushroom stains. The brown sugar selected out the carbon source of the agricultural medium. Also the soybean flour selected out the nitrogen source of agricultural medium. In the medium selection, we selected out agricultural optimum medium composed of brown sugar 3%, soybean flour 0.3%, potassium phosphate 0.05%, magnesium sulfate 0.05%. Under the 250 ml erlenmeyer culture, the effects of such factors as the inoculum rate, the working volume, cultural method and flask shapes on the mycelial growth were examined. The optimal inoculum rate and working volume on the mycelial growth of oyster mushroom was 2 mycelial disk (diameter 6 mm) and 100 ml, respectively. The shake flask culture was enhanced the mycelial growth than at the erlenmeyer flask. Pulp form growth of mycelium in the erlenmeyer flask culture was obtained in the culture with glass rod of length 50 mm, diameter 10 mm.

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Study on Polysaccharide Production with Paecilomyces japonica in Flask Culture

  • Park, Seok-Jae;Han, Dae-Seok;Hong, Eok-Gi
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.235-238
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    • 2000
  • The production of polysaccharide from Paecilomyces japonica was studied in the shake flask culture. For the cell growth and the polysaccharide production, the optimum synthetic medium was glucose peptone(YMP) medium. The flask culture conditions for the polysaccharide production were $27^{\circ}C$ and 200rpm with the initial pH 9 for 8days cultivation.

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Effects of Environmental and Nutritional Conditions on Fibrinolytic enzyme Production from Bacillus subtilis BK-17 in Flask Culture (플라스크 배양에서 Bacillus subtilis BK-17의 혈전용해효소 생산에 대한 환경 및 영양 조건의 영향)

  • 최원아;이진욱;이경희;박성훈
    • KSBB Journal
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    • v.13 no.5
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    • pp.491-496
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    • 1998
  • The production of fibrinolytic enzyme from Bacillus subtilis BK-17 was studied in the shake flask cultures. The important medium components studied include nitrogen source, carbon source and inorganic salts. The environmental conditions include initial pH, temperature, shaking speed and working volume. Among various N-sources, C-sources and inorganic salts tested, soybean flour, D-glucose and Na2HPO4 gave the best results, and their optimal concentrations were 1.5%, 0.5% and 0.05%, respectively. The optimal pH and temperature were 9.0 and 37$^{\circ}C$. With decreasing working volume in the range of 25∼100ml in the 250ml flask or increasing shaking speed in the range of 100∼300rpm, the enzyme production was greatly enhanced. The enzyme activity under the optimal conditions was about 1400I.U./ml with urokinase as a standard.

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Effect of Glycine on L-Ornithine Production by a Citrulline Auxotroph of Brevibacterium ketoglutamicum and Stoichiometric Analysis

  • Nam, Soo-Wan;Choi, Dae-Keon;Ryu, Wuk-Sang;Jang, Hyung-Wook;Chung, Bong-Hyun;Park, Young-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.4 no.2
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    • pp.95-101
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    • 1994
  • The effects of glycine on cell growth and L-omithine production were investigated in shake-flask and jar fermentor cultures of a citrulline auxotrophic mutant, Brevibacterium ketoglutamicum BK 1046. In the shake-flask culture, the optimal concentration of glycine for L-ornithine production was found to be 20 g/l. In the jar fermentor culture with the glycine at an initial concentration of 20 g/l, L-ornithine production increased by 28%, compared to that of the culture with no glycine added. 37 g/l of L-ornithine was produced when additional feeding of glycine (5 g/l) was made. This was a significant improvement in L-ornithine production compared to that (ca. 24 g/l) of the corresponding batch culture conducted without glycine. According to the stoichiometric analysis with the batch fermentation results, the experimental and theoretical L-ornithine yields based on the glucose consumption were 0.24 and 0.59, respectively. This indicates that the performance of L-ornithine fermentation can further be improved by the supplementation of glycine and the development of a mutant strain possessing a higher growth yield.

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Improved Astaxanthin Production of Xanthophyllomyces dendrorhous with Carotenogenesis Stimulating Factors (Carotenogenesis의 생성 자극 인자를 이용한 Xanthophyllomyces dendrorhous에 의한 astaxanthin 산물의 증가)

  • Xu, Bao-Jun;Sung, Chang-Keun;Li, Chang-Tian;Mo, Eun-Kyoung
    • Journal of Life Science
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    • v.14 no.3
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    • pp.472-477
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    • 2004
  • To improve biomass and astaxanthin production by wild-type Xanthophyllomyces dendrorhous simultaneously in shake flask culture, physical factors, nutritional factors and carotenogenesis stimulating factors affecting astaxanthin production were studied on base of HPLC quantitative analysis. The results suggested that carotenogenesis precursor composition acetic acid, mevalonic acid, tomato extract, and carrot extract could increase the productivity of astaxanthin markedly based on the optimized temperature, initial pH value, carbon and nitrogen sources conditions.

Antibacterial Activity by Chitosan Derivatives with Quaternary Ammonium Salt (4차 암모늄기를 가진 키토산 유도체의 항균성)

  • Kim, C.H.;Choi, Y.S.;Choi, K.S.
    • Applied Chemistry for Engineering
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    • v.7 no.5
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    • pp.1020-1026
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    • 1996
  • Chitosan is the deacetylation product of chitin. Chitosan was converted to N-alkyl chitosan derivatives using proper aldehyde having different alkyl chains, then chitosan derivatives with quaternary ammonium salt were producted by quaternization with methyl iodide. Their antibacterial activities aganst S. aureus(ATCC 6538P) and E. coli (ATCC 14339), gram-positive bacteria and gram-negative bacteria, respectively, as introduced alkyl length were evaluated by shake flask method using colony count. The antibacteiral activity was found to be increased as alkyl chains. hydrophobic goups. These antibacterial agents were more active against S. aureus than E. coli. It may be due to the different of structure of cytoplasmic membrane.

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Morphological and rheological properties of culture broth of Cephalosporium acremonium M25

  • Lim, Jung-Soo;Kim, Jin-Hee;Kim, Chongyoup;Kim, Seung-Wook
    • Korea-Australia Rheology Journal
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    • v.14 no.1
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    • pp.11-16
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    • 2002
  • Cephalosporium acremonium is a filamentous microorganism producing cephalosporin C. The morphological differentiation of C. acremonium in submerged culture is closely related with the rheological properties of culture broth and production of cephalosporin C. In this study, the rheological and morphological properties of culture broth of C. acremonium were investigated. In the seed broths of shake-flask and fermenter culture, the Herschel-Berkley equation was in excellent agreement with experimental results in the whole range of shear rate. In the seed broths of shake-flask culture, morphological differentiation into arthrospores affected to changes of apparent viscosity. But results in the fermenter culture, morphological factors such as mean hyphal thickness and the number of tips gave more effect on changes of apparent vitacosity than differentiation into arthrospores. Overall, it suggested that the morphological parameters measured by image analysis can be used as a good parameter to indicate the rheological properties of culture broth of C. acremonium M25.