• Title/Summary/Keyword: Shake culture

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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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The Improvement of Cephalosporin C Production by Fed-batch Culture of Cephalosporium acremonium M25 Using Rice Oil

  • Kim Jin Hee;Lim Jung Soo;Kim Seung Wook
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.6
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    • pp.459-464
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    • 2004
  • The objective of this study is to improve cephalosporin C (CPC) production byoptimization of medium and culture conditions. A statistical method was introduced to optimize the main culture medium. The main medium for CPC production was optimized using a statistical method. Glucose and corn steep liquor (CSL) were found to be the most effective factors for CPC production. Glucose and CSL were optimized to 2.84 and $6.68\%$, respectively. CPC produc­tion was improved $50\%$ by feeding of $5\%$ rice oil at day 3rd and 5th day during the shake flask culture of C acremonium M25. The effect of agitation speeds on CPC production in a 2.5-L bio­reactor was also investigated with fed-batch mode. The maximum cell mass (54.5 g/L) was obtained at 600 rpm. However, the maximum CPC production (0.98 g/L) was obtained at 500 rpm. At this condition, the maximum CPC production was improved about $132\%$ compared to the re­sult with batch flask culture.

Production of Rosmarinic Acid, Lithospermic Acid B, and Tanshinones by Suspension Cultures of Ti-Transformed Salvia miltiorrhiza Cells in Bioreactors

  • Zhong, Jian-Jiang;Hui Chen;Feng Chen
    • Journal of Plant Biotechnology
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    • v.3 no.2
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    • pp.107-112
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    • 2001
  • The kinetics of Ti-transformed Salvia miltiorrhiza cell cultures was studied in 250-$m\ell$ shake flasks by using B5 medium with addition of 30 gfL of sucrose. In the cell cultures, the maximum cell mass obtained was 11.5 g DW/L on day 15. The highest amount of phenolic compounds - rosmarinic acid (RA) and lithospermic acid B (LAB) reached 871.3 mg/L (day 15) and 121.3 mg/L (day 13), respectively. The total tanshinone production, i.e., intracellular plus extracellular cryptotanshinone, tanshinone 1, and tanshinone IIA, was 5.3 mg/L on day 13. For the cultivations in 2.4-L stirred bioreactors, the residual sugar level and medium conductivity were a little higher in a small turbine impeller reactor ($T_s$) than those in a large turbine impeller reactor ($T_L$), while a higher cell density was obtained in the $T_L$. For the production of tanshinones and phenolics, better results were obtained in the $T_L$ than in the $T_s$. In the $T_L$, similar or even a little higher production titers of tanshinones and phenolic compounds were achieved compared to those in the flasks. The results suggest that the shake flask results could be successfully scaled up to the $T_L$ reactor. Such a large impeller reactor like $T_L$ may be better than a small impeller one for the large-scale production of the valuable metabolites by the suspension cultures of Ti transformed S.miltiorrhiza cells. This is considered due to the beneficial culture environment in the $T_L$, such as low shear rates as estimated theoretically.

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Spore Production of Entomopathogenic Fungus, Beauveria bassiana 726, Using Molasses (당밀을 이용한 곤충병원성 곰팡이 Beauveria bassiana 726의 포자생산)

  • 김병혁;강성우;윤철식;성재모;홍석인;김승욱
    • KSBB Journal
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    • v.14 no.3
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    • pp.365-370
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    • 1999
  • To optimize the culture conditions for Beauveria bassiana 726, the effects of culture medium, pH, and temperature on mycelium and spore production were investigated. The optimum temperature and pH for the cultivation of B. bassiana 726 were 28 $^{\circ}C$ and 5.0, respectively. The optimized medium was composed of 1.0~2.0% total sugar from molasses, 0.5% corn steep liquor and 0.05% KH$_2$PO$_4$. In the cultivation of B. bassiana 726 with the optimum medium, the specific growth rate and substrate utilization were well-fitted with the proposed kinetic model in the shake flask and stirred tank reactor. When the fed-batch cultivation using carbon suorce, nitrogen source, and mineral salt as a feeding medium was compared with batch cultivation in stirred tank reactor, mycelium (12.7 g/L) and spore production (5.4$\times$$10^8/mL$) were enhanced up to 110% and 85%, respectively.

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Bioflocculant production from Bacillus sp. A56 (Bacillus sp. A56을 이용한 응집제 생산)

  • 서현효;이문호;김희식;박찬선;윤병대
    • Microbiology and Biotechnology Letters
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    • v.21 no.5
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    • pp.486-493
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    • 1993
  • A gram(+) bacteria that produced microbial flocculant was isolated from soil and classified as a Bacillus species and named as Bacillus sp. A56. The culture conditions of the strain for fluocculant production were studied in a shake flask. Optimum temperature and initial pH for flcculant production were 30C and 6.5, respectively. The optimized medium has gollowing composition: glucose 20g/l, NH4NO3 0.5g/l, K2HPO4 1.0g/l, KH2PO4 0.8g/l, MgSO4.7H2O 0.2g/l, MnSO4.4-6H2O 0.3g/l, CaCO3 0.5g/l, yeast extract 0.3g/l, tryptone 0.3g/l in tap water.

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Studies on the Antibiotic Constituents of Korean Basidiomycetes(IV). Preliminary examination of the mycelial cultures of the 17 basidiornycetous strains

  • Chung, Kyeong-Soo;Chung, Soo-Hyun;Kim, Byong-Kak
    • Archives of Pharmacal Research
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    • v.11 no.1
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    • pp.61-64
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    • 1988
  • To investigate the antibiotic constituents of Korean basidiomycetes the carpophores of the wood-rotting basidiomycetes were collected from several locations of Korea, and from them 17 mycelial strains were isolated on potato-dextrose-agar plates supplemented with tetracycline ($20\;{\mu}g/m{\ell}$. The isolated strains were shake-cultured in glucosepeptone-yeast extract medium and then the antibacterial activities of the culture filtrates were assessed by disc-plate method. Among them, 12 strains (70.6%) were active, and basidiomycete strain LMCB-109 (Daedalea quercina) and LMCB-116 showed potent activities against all the six bacterial target organisms including Serratia marcescens.

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Optimal Culture Conditions for Mycelial Growth and Exo-polymer Production of Ganoderma applanatum

  • Jeong, Yong-Tae;Jeong, Sang-Chul;Yang, Byung-Keun;Islam, Rezuanul;Song, Chi-Hyun
    • Mycobiology
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    • v.37 no.2
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    • pp.89-93
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    • 2009
  • The effect of fermentation parameters and medium composition on the simultaneous mycelial growth and exo-polymer production from submerged cultures of Ganoderma applanatum was investigated in shake-flask cultures. The optimum initial pH for mycelial growth and exo-polymer production was 5.0 and 6.0, respectively. The optimum temperature was $25^{\circ}C$ and the optimum inoculum content was 3.0% (v/v). The optimal carbon and nitrogen sources were glucose and corn steep powder, respectively. After 12 days fermentation under these conditions, the highest mycelial growth was 18.0 g/l and the highest exo-polymer production was 3.9 g/l.

Effect of Levulinic Acid on the Production of Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) by Ralstonia eutropha KHB-8862

  • Chung, Sun-Ho;Park, Gang-Guk;Kim, Hyung-Woo;Rhee, Young-Ha
    • Journal of Microbiology
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    • v.39 no.1
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    • pp.79-82
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    • 2001
  • The influence of levulinic acid (LA) on the production of copolyester consisting of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV) by Ralstonia eutropha was investigated. Addition of LA into the culture medium greatly increased the molar fraction of 3HV in the copolyester, indicating that LA can be utilized as a precursor of 3HV. In shake flask culture, the 3HV content in the copolyester increased from 7 to 75 mol% by adding 0.5 to 4.0 g/L LA to the medium containing fructose syrup as a main carbon source. A maximal copolyester concentration of 3.6 g/L (69% of dry cell weight) was achieved with a 3HV content of 40 mo1% in a jar fermentor culture containing 4.0 g/L of LA. When LA (total concentration, 4 g/L) was added repeatedly into a fermentor culture to maintain its concentration at a low level, the copolyester content and the 3HV yield from LA reached up to 85% of dry cell weight and 5.0 g/g, respectively, which were significantly higher than those when the same concentration of the LA was supplied al1 at once. The present results indicated that LA is more effective than propionate or valerate as a cosubstrate fur the production of copolyesters with varying molar fractions of 3HV by R. eutropha.

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Expression and Secretion of Human Serum Albumin in the Yeast Saccharomyces cerevisae

  • Kang, Hyun-Ah;Jung, Moon-Soo;Hong, Won-Kyoung;Sohn, Jung-Hoon;Choi, Eui-Sung;Rhee, Sang-Ki
    • Journal of Microbiology and Biotechnology
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    • v.8 no.1
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    • pp.42-48
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    • 1998
  • In order to maximize the secretory expression of human serum albumin (HSA) in the yeast Saccharomyces cerevisiae, a series of HSA expression vectors were constructed with a combination of different promoters, 5' untranslated regions (5'UTR), and secretion signal sequences. The expression vector composed of the galactose-inducible promoter GALl0, the natural 5'UTR, and the natural signal sequence of HSA directed the most efficient expression and secretion of HSA among the constructed vectors when introduced into several S. cerevisiae strains. Although the major form of HSA expressed and secreted in the yeast transformants was the mature form of 66 kDa, the truncated form of 45 kDa was also detected both in the cell extract and in the culture supernatant. The level of the intact HSA protein in the culture supernatant reached up to 30 mg/l at 24 h of cultivation in a shake-flask culture but began to decrease afterwards, indicating that the secreted HSA protein was unstable in a prolonged culture of yeast.

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Effect of Galactose and Dextrose on Human Lipocortin I Expression in Recombinant Saccharomyces cerevisiae Carrying Galactose-Regulated Expression System

  • Nam, Soo-Wan;Seo, Dong-Jin;Rhee, Sang-Ki;Park, Young-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.3 no.3
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    • pp.168-173
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    • 1993
  • The expression kinetics of human lipocortin I (LCI), a potential anti-inflammatory agent, was studied in the shake-flask and fermenter cultures of Saccharomyces cerevisiae carrying a galactose-inducible expression system. The cell growth, expression level of LCI, and the plasmid stability were investigted under various galactose induction conditions. The expression of LCI was repressed by the presence of a very small amount of dextrose in the culture medium, but it was induced by galactose after dextrose became completely depleted. The optimal ratio of dextrose to galactose for lipocortin I production was found to be 1.0 (10 g/l dextrose and 10 g/l galactose). With optimal D/G ratio of 1.0 and the addition of galactose prior to dextrose depletion, LCI of about 100~130 mg/l was produced. LCI at a concentration of 174 mg/l was porduced in the fed-batch culture, which was nearly a twice as much of that produced in the batch culture. The plasmid stability was very high in all culture cases, and thus was considered to be not an important parameter in the expression of LCI.

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