• Title/Summary/Keyword: Shaking flask culture

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High-Yield Production of Astragalosides from Transgenic Hairy Root Cultures of Astragalus membranaceus (형질전환된 황기 모상근으로부터 Astragalosides의 생산을 위한 연구)

  • Hwang, Sung-Jin
    • KSBB Journal
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    • v.21 no.2
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    • pp.123-128
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    • 2006
  • A transgenic hairy root clone AG-04 of Astragalus membranaceus was obtained following co-cultivation of leaf explants with Agrobacterium rhizogenes ATCC15834. This clone was examined for its growth and production of cyclolanostane-type saponins, astragalosides I, II, and III, under various culture conditions. Among the five basal media tested, Shenk and Hildebrandt(SH)(18) medium was best for roots growth and astragalosides production. The maximum root biomass was obtained at inoculum size of 500 mg FRW per flask, initial sucrose concentration of 3%, and shaking speeds of 90 rpm. The astagalosides production was promoted when the hairy root clone AG-04 was cultured at shaking speeds of 120 rpm and light irradiation of 18 h. Astragaloside contents was also stimulated with high initial sucrose concentration, and the maximum astargalosides contents of 6.21 %/g DRW was obtained at initial sucrose concentration of 6%. The addition of chitosan(100 mg/L) to the culture medium was significantly increased astragalosides production. This was 2.1 times higher than that obtained in a control culture without chitosan.

Studies on the Hydrolysis of Inulin in Jerusalem Artichokes by Fungal Inulase (미생물(微生物) Inulase에 의(依)한 돼지감자 중의 Inulin분해(分解)에 관한 연구(硏究))

  • Kim, Ki-Choul
    • Applied Biological Chemistry
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    • v.18 no.3
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    • pp.177-182
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    • 1975
  • The analysis of Jerusalem artichoke showed that it contains 12.09% of Inulin. The results obtained from the examination of the conditions for fructose production by cultivating Pencillum sp 1 in the Jerusalem articoke medium were as follows: 1. The optimum amount of water added to Jerusalem artichoke was 2.5 $\ell$ of distilled water per ㎏ of fresh Jerusalem artichoke. It this case, the concentration of Inulin was 4% (w/v). 2. The optimum temperature was $30^{\circ}C$, the initial optimum pH was 5.0 and the optimum cultural period was 72 hours. 3. Shaking culture with 50 ml of the medium and 120 oscills/min in 500 ml shaking flask was most effective as the culture method. 4. 0.1% of $NH_4H_2PO_4$ as a nitrogen source, 0.001 of $FeSO_47H_20$ and 0.001% of $MgSO_47H_2$ as metal salts were most effective. 5. Fructose production continued to increase for 72 hours under the optimum conditions for cultivation and the highest production rate to the Inulin was 95.25%.

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Detection of Lignans from Transformed Root Cultures of Schisandra chinensis Baillon (오미자의 형질전환된 근으로부터 리그난 화합물의 검출)

  • Hwang, Sung-Jin;Pyo, Byoung-Sik;Hwang, Baik
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.6
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    • pp.448-453
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    • 2004
  • Transformed roots of Schisandra chinensis were obtained following co-cultivation of in vitro cultivated plantlet segments with Agrobaterium rhizogens ATCC15834. This root was examined for its growth and gomisin J contents under various culture conditions. Among the six basal culture media tested, WPM (Lloyd & McCown, 1980) medium supplemented with 5% sucrose was the best roots growth 6.2 (g D.W/flask) and gomisin J accumulation 1.56 $(X10^{-3}\;ug/g\;D.W)$. Initial inoculum size correlated with the yield of biomass while gomisin J contents was not affect. Gomisin J production was influenced by the initial sucrose concentration and the highest production yield was achieved at the concentration of 7%. The optimal shaking speeds for roots growth and gomisin J production was 120 and 140 rpm, respectively.

Degradation of Fats, Oils and Hydrocarbons by Acinetobacter calcoaceticus (Acinetobacter calcoaceticus에 의한 유지와 탄화수소의 분해)

  • 고정삼;고영환;김권수;양상호;강경수
    • Microbiology and Biotechnology Letters
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    • v.20 no.4
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    • pp.477-482
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    • 1992
  • A bacterial strain Acinetobacter calcoaceticus was examined for its ability to degrade fats, oils and hydrocarbons, and tested for the possibility of application in wastewater treatment. All fats and oils tested were degraded by the strain. About 60% of hexadecane, 26% of fish oiL and 40-54% of vegetable oils were consumed respectively in shaking-flask culture. Saturated fatty acid compositions were about 55% in fish oil and 6-12% in vegetable oils. Increases in cell mass were accompanied with decreases in the concentrations of carbon sources. When jar fermentor in place of shaking-flask was used as a culturing vessel. above 80% of all carbon sources was consumed and yield of cell mass was improved to nearly 1.00. Synthetic wastewaters containing 3% of fat, oil, or hydrocarbon as a sale ca,bon source were treated sequentially with A. calcoaceticus first and then exposed to activated sludge. The concentrations of carbon sources were decreased below 0.06% through the process, and the concentrations of suspended solids were lower than 53 mglml. The data imply the potential use of A. calcoaceticus in wastewater treatment.

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Distribution and Substrate Specificity of 5-fluorocytosine Deamiase in Bacteria (세균의 5-Fluorocytosine Deaminase의 분포와 기질 특이성)

  • 전홍기;김동완
    • Microbiology and Biotechnology Letters
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    • v.13 no.4
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    • pp.361-366
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    • 1985
  • Distribution and substrate specificity of 5-fluorocytosine deaminase were studied in various genera of bacteria. 5-Fluorocytosine deaminase was produced by various bacteria independent of genus and species and it catalyzed the deamination of cytosine, 5-fluorocytosine and 5-methylcytosine. Xanthomonas campestris IAM 1671 produced relatively large amount of 5-fluorocytosine deaminase. The composition of optimum culture medium for enzyme production wat glycerine 0.5%, peptone 1%, yeast extract 0.5%, NaCl 0.5% and the initial pH of the medium was 7.5. The highest enzyme formation was observed after 24 hours of cultivation In 500$m\ell$ shaking flask containing 90$m\ell$ of medium at 3$0^{\circ}C$ on a reciprocal shaker.

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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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Optimization of Culture Conditions for the [+]-Eudesmin Production in Magnolia Sieboldii Cells (함박꽃나무의 현탁배양세포로부터 [+]-Eudesmin의 생산을 위한 최적화)

  • Hwang Sung Jin
    • KSBB Journal
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    • v.20 no.1 s.90
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    • pp.34-39
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    • 2005
  • In order to product the furofuranoid lignans, (+)-eudesmin which is one of the secondary products from Magnolia sieboldii. through cell suspension cultures; various culture media, initial sucrose concentration, elicitations, shaking speeds, and inoculum sizes. Among the culture media tested, MS medium had a pronounced effect on suspension cell growth and (+)-eudesmin contents. The maximum dry cell weight (DCW) of 3.71 g per flask was obtained at inoculum size of 0.5 g and in MS medium supplemented with $3\%$ sucrose plus 0.5 mg/L 2,4-D after 8 weeks. (+)-Eudesmin biosynthesis was stimulated with high initial sucrose concentration ,and the maximum (+)-eudesmin production of $3.2{\mu}g/g$ DCW was achieved at 200mg/L chitosan and $5\%$ initial medium sucrose. The optimal shaking speeds for dry biomass accumulation and (+)-eudesmin contents was 130 rpm. This work is considered to be helpful for large-scale bioprocessing of Magnolia sieboldii suspension cell cultures in bioreactor.

Formation and Production of Ascorbate Oxidase by Cucumber Tissue Cultured Cells (오이조직 배양세포에 의한 Ascorbate Oxidase 생성 및 생산)

  • Lee, Jong-Hwa;Chung, Ho-Kwon;Shinmyo, Atsuhiko;Lim, Bun-Sam
    • Microbiology and Biotechnology Letters
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    • v.21 no.4
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    • pp.329-335
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    • 1993
  • Ascorbate oxidase activity in various cucumber tissue extracts was highest in young fruit peeling. Cucumber callus was induced from young fruit peeling and callus cell lines were selected for more than 7 months, which porduced high levels of ascorbate oxidase and had a high growth rate. Induction of callus was optimized with Linsmaier-Skoog(LS) medium at 25$^{\circ}C$ in dark phase. Ascorbate oxidase activity reached a maximum at 5 days after transfer to LS basal liquid-medium ant then declined. The enzyme activity in callus cells was stimulated by addition of 10${\mu}$M $CuSO_4$ in the early logarithmic phase of growth. And also, adding 10${\mu}$M $CuSO_4$ at 3rd day 7th day of culture period, ascorbate oxidase activity in callus cells was maintained to high level. Maximum yield of ascorbate oxidase was found at the 25th day by flask shaking culture, but three-fold of ascorbate oxidase activity was obtained at the 16th day by jar fermentation.

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Optimal Culture Conditions for Production of Environment-Friendly Biosurfactant by Pseudomonas sp. EL-G527 (Pseudomonas sp. EL-G527에 의한 환경친화성 생물계면활성제의 생산최적조건)

  • 차미선;임은경;이근희;조순자;손홍주;이상준
    • Journal of Environmental Science International
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    • v.11 no.3
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    • pp.177-182
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    • 2002
  • A biosurfactant-producing microorganism was isolated from activated sludge by enrichment culture when grown on a minimal salt medium containing n-hexadecane as a sole carbon source. This microorganism was identified as Pseudomonas sp. and it was named Pseudomonas sp. EL-G527. It's optimal culture condition is 2% n-hexadecane, 0.2% NH$_4$NO$_3$, 0.3% KH$_2$PO$_4$, 0.3% $K_2$HPO$_4$, 0.02% MgSO$_4$ㆍ7$H_2O$, 0.0025% CaCk$_2$ㆍ6$H_2O$, 0.0015% FeSO$_4$ㆍ7$H_2O$ in 1$\ell$ distilled water and initial pH 7.0. Cultivation was initiated with a 2% inoculum obtained from starter cultures grown in 30 $m\ell$ of the same medium in 250 $m\ell$ flask. They were cultivated at 3$0^{\circ}C$ in reciprocal shaking incubator and the highest biosurfactant production was observed after 4 days.

Isolation and Characterization of Phosphorus Accumulating Acinetobacter CW3 (인 축적균 Acinetobacter CW3의 분리 및 특성)

  • 심성훈;류원률;이영호;김정목;조무환
    • KSBB Journal
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    • v.14 no.1
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    • pp.71-75
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    • 1999
  • A highly effective phosphorus accumulating bacterium named Acinetobacter CW3 was isolated from the nature by using Winogradsky columns. The optimal cultivation conditions of Acinetobacter CW3 in shaking flask were determined as $20^{\circ}C$, pH 7, 200rpm, 18.5mg $PO_4$-P/L. Acientobacter CW3 could remove phosphorus completely in 12hours for a batch culture at optimal cultivation condition. This bacterium could uptake phosphorus on aerobic condition and release it on anaerobic condition.

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