• Title/Summary/Keyword: Agitation culture

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Selection of organic Nitrogen Source and Optimization of Culture Conditions for the Production of Arachidonic Acid from Mortierella alpina (Mortierella alpina를 이용한 아라키돈산의 생산에서 유기질소원의 선정과 배양 조건의 최적화)

  • 유연우;하석진;박장서
    • KSBB Journal
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    • v.19 no.1
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    • pp.78-82
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    • 2004
  • Experiments were carried out to select an organic nitrogen source and optimize the culture conditions for the production of arachidonic acid by Mortierella alpina DSA-12. Corn steep powder(CSP) was selected as an organic nitrogen source based on arachidonic acid production and raw material price. The optimum C/N ratio was in the range of 15 to 17 with the medium containing glucose as carbon source and CSP as nitrogen source. The optimum culture conditions for arachidonic acid production showed 500 rpm agitation and 25$^{\circ}C$ culture temperature at 0.5 vvm aeration. Under the optimum conditions, the concentration of cell, total lipid and arachidonic acid were 21.8 g/L, 10.2 g/L and 3.70 g/L, respectively, from 50 g/L glucose and 18 g/L CSP. In the 500 L fermenter with 0.5 vvm aeration and 200 rpm agitation, the concentration of cell, total lipid and arachidonic acid were 19.8 g/L, 9.1 g/L and 3.67 g/L, respectively, from 50 g/L glucose and 18 g/L CSP. This result showed that an arachidonic acid production could be possible with a bench-scale fermenter using corn steep powder as a nitrogen source.

Control of Both Foam and Dissolved Oxygen in the Presence of a Surfactant for Production of $\beta$-Carotene in Blakeslea trispora

  • Kim, Seon-Won;Lee, In-Young;Jeong, Jae-Cheol;Lee, Jung-Heon;Park, Young-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.548-553
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    • 1999
  • A production of $\beta-Carotene$was attempted in a fed-batch culture of Blakeslea trispora by controlling both foam and dissolved oxygen in the presence of surfactant, Span 20. Results obtained from the shake flask cultures indicated that a high concentration of dissolved oxygen was needed for both cell growth and $\beta-Carotene$ synthesis, and the optimal concentration of glucose was found to be in the range of 50-100 g/l. In order to maintain the dissolved oxygen concentration level at higher than 50% of air saturation, pure oxygen was automatically sparged into the medium with air. Foam was controlled by bypassing air from the submerged aeration to the headspace in response to the foam that was caused by Span 20. High agitation speed was found to be detrimental to the cell growth due to shear damage, even though it provided sufficient dissolved oxygen. On the other hand, a low aeration speed caused stagnant regions in the fermentor because of improper mixing. Thus, for the fed-batch operation, agitation speed was increased gradually from 300 to 700 rpm to prevent cell damage at the initial stage of fermentation and to give efficient mixing for a viscous culture broth as the culture proceeded. By controlling dissolved oxygen and foam, a high concentration of $\beta-Carotene$otene (1,190 mg/l) was obtained in 6 days of the fed-batch culture of B. trispora with 2.5% of the dry cell weight, which was approximately 5 times higher than that of the batch cultures.

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Effects of Culture Conditions of Rhizopus sp. ZB9 on the Production of Protease during Preparation of Rice Koji (쌀 입국 제조시 Rhizopus sp. ZB9의 배양 조건이 프로테아제 생성에 미치는 영향)

  • So, Myung-Hwan;Lee, Young-Sook
    • The Korean Journal of Food And Nutrition
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    • v.23 no.3
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    • pp.399-404
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    • 2010
  • This study was conducted to determine the influence of culture conditions such as temperature, time, water content, koji-thickness, and agitation on the production of protease by Rhizopus sp. ZB9, isolated from Korean Nuruk, during the preparation of rice koji, which is used in brewing the Korean rice wines, Takju and Yakju. Rice koji was made under different culture conditions, and the proteolytic activity of each koji was tested. The temperature range suitable for the production of protease was $28~32^{\circ}C$. Based on the protease and color, 60 hours of cultivation at $28^{\circ}C$ was shown to produce optimum results. The production of protease increased in proportion to the increase in water content of steamed rice from 25% to 35%. An increase in koji-thickness induced no adverse effects on the production of protease, and agitation during cultivation showed beneficial effects.

Effect of Culture Conditions on Microbial Cellulose Production by Acetobacter sp. A9 in Shaking Cultures

  • Son, Hong-Joo;Kim, Han-Soo
    • Journal of Life Science
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    • v.11 no.1
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    • pp.11-13
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    • 2001
  • Several culture conditions affecting cellulose production by a newly isolated Acetobacter sp. A9 were examined by cultivating cells under shaking cultures. The inoculum size in the range of 1-10% (v/v) did not influence cellulose production. Maximum cellulose production was obtained with 200 rpm of agitation speed. The cells grown in the 75 ml of medium in a 250-ml conical flask produced the highest level of cellulose. The strain was able to produce cellulose at 25-3$0^{\circ}C$ with a maximum at 3$0^{\circ}C$. Cellulose production occurred at pH 4.5-7.5 with a maximum at pH6.5.

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Amylase Production by Continuous Cultures of Aspergillus oryzae and its Mutants (Aspergllus oryza와 그 변이주의 연적배양에 의한 amylase 생성에 관한 연구)

  • Han, Hong-Eui
    • Korean Journal of Microbiology
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    • v.15 no.2
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    • pp.63-76
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    • 1977
  • Irradiation with high doses of gamma rays induced the reduction of mycelial weight and anaylase activity, and increased relative amylase activity in surface cultures. Biphase in growth curves was shown in aeration-agitation cultures but the behavior of the first phase of growth could be eliminated by replacing the amylasehydrolysed starch substrates, so that enzyme production was shortened ca. 40 hours and relative amylase activity was increased about 3 times higher before onset of autolysis. In the effect of gibberellin on amylase production, the positive stimulation was appeared to only surface culturs of the liquid medium and the negative effect to shake-cultures in a mutant. Trials of various continuous culture were resulted not only the approalch to the value of amylase activity in surface cultures of liquid medium, but also higher productivity than in batch cultures. The culture-degeneration was observed in two-stage continuous culture, but did not appear in continuous elevation culture.

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In Vitro Proliferation Model of Helicobacter pylori Required for Large-Scale Cultivation

  • Oh, Heung-Il;Lee, Heung-Shick;Kim, Kyung-Hyun;Paek, Se-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.10 no.3
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    • pp.367-374
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    • 2000
  • The composition of dissolved gases and nutrients in a liquid medium were determined for establishment of the optimum conditions for in vitro culture of Helicobacter pylori. A microaerobic condition facored by the organism was prepared by adjusting the partial pressure of the gas, agitation speed, and viscosity of the medium. The gaseous concentrations were controlled by utilizing CampyPak Plus that reduced oxygen while augmenting carbon dioxide. Agitation of the broth facilitated the oxygen transfer to the cells, yet inhibited the growth at high rates. An increase of viscosity in the medium repressed the culture although this variable was relatively insignificant. The chemical constituents of the liquid broth were examined to establish an economic model for H. pylori cultivation. The microbe required a neutral pH for optimum growth, and yet was also able to proliferate in an acidic condition, presumably by releasing the acidity-modulating enzyme, urease. Cyclodextrin and casamino acid were investigated as growth enhancers in place of serum, while yeast extract unexpectedly inhibited the cells. A low concentration of glucose, the unique carbon source for the organism, increased the cell density, yet high concentrations resulted in an adverse effect. Under optimally dissolved gas conditions, the cell concentration in brucella broth supplemented with serum substitutes and glucose reached $1.6{\times}10^8$ viable cells/ml which was approximately 50% higher than that obtained in the liquid medium added with only cyclodextrin or serum.

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Liquid culture condition of Tremella fuciformis mycelia (흰목이 균사 액체배양 조건)

  • Chang, Hyun-You;Lee, Chan;Choi, Sung-Woo;Yun, Jong Won
    • Journal of Mushroom
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    • v.6 no.1
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    • pp.27-31
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    • 2008
  • The optimization of submerged culture conditions for mycelial growth and exopolysaccharide (EPS) production in an edible mushroom Tremella fuciformis were studied in shake flasks and bioreactors. The temperature of $28^{\circ}C$ and pH 8 in the beginning of fermentation in agitated flasks was the most efficient condition to obtain maximum mycelial biomass and EPS. The optimal medium constituents were as follows (g l-1): glucose 20, tryptone 2, $KH_2PO_4$ 0.46, $K_2HPO_4$ 1 and $MgSO_4H_2O$ 0.5. The fungus was cultivated under various agitation and aeration conditions in a 5L stirred-tank bioreactor. The maximum cell mass and EPS production were obtained at a relatively high agitation speed of 200 rpm and at an aeration rate of 2 vvm. The flow behavior of the fermentation broth was Newtonian and the maximum apparent viscosity (35 cP) was observed at a highly aerated condition (2 vvm). The EPS productivity in an airlift reactor was higher than that in the stirred-tank reactor. The EPS was protein-bound polysaccharides consisted of mainly mannose, xylose, and fructose. The molecular weights of EPS were determined to be $1.3{\sim}1.5{\times}10^6$.

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High Cell Density Culture of Micro-algal Dunaliella bardawil (미세조류 Dunaliella bardawil의 고농도 세포배양)

  • 정욱진;왕만식;최승인;정병철;김주곤
    • KSBB Journal
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    • v.14 no.2
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    • pp.160-166
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    • 1999
  • High cell density cultivation of microalga Dunaliella bardawil using nitrogen fed-batch cultures was studied in batch flask. Optimum environmental conditions include concentrated nutrients except NaCl and carbon sources, carbon sources, pH, light, agitation, nitrate and phosphate ions. Cell growth, consumption rates of nitrate and phosphate ions were monitored. Optimal conditions for higher cell density were found to be(in the range tested): 5 times concentrated media(1 times-10 times concentrated media) pH 8.0 (7.0-9.0) white light(blue and red light) 15mM of nitrate (0.94-15mM) 250mM $NaHCO_3$ and $CO_2$ gas. However, the addition of phosphate ions did not enhance the algal maximum cell density and specific growth rate. Nitrate was found to be effective for the cell growth. The maximum cell density of fed-batch culture using nitrate ions in $8.955{\times}106$cells/ml after 189hr incubation.

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Effect of Agitation and Aeration Rate on Nicotiana tabacum Suspension Cell Culture in Bioreactors (Bioreactor를 이용한 담배세포 현탁배양에서 교반형태와 통기량이 미치는 영향)

  • Lee, Sang-Yun;Kim, Dong-Il
    • KSBB Journal
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    • v.14 no.5
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    • pp.534-538
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    • 1999
  • For the optimization of operating conditions for plant cell suspension culture in bioreactors, effects of bioreactor types, various kinds of impellers, and aeration rates were examined using Nicotiana tabacum cells as a model system. Stirred tank bioreactor and airlift bioreactor were used for the comparison of bioreactor type. Growth rates in both bioreactors were lower than in shake flasks. In terms of final cell concentration, stirred tank bioreactor supported a little bit better growth compared to airlift bioreactor. Impeller type did not affect cell growth significantly, but it was apparent that cell size index decreased in the case of using hollowed paddle impeller. When the aeration rate was maintained at 0.3 vvm, cell growth was the best. At above 1.0 vvm, growth inhibition as well a browning was noticed. In addition, it was found that cell size index reduced proportionally to the increased of aeration rate.

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Effects of Dissolved Oxygen Level on Avermectin $B_{1a}$ Production by Streptomyces avermitilis in Computer-Controlled Bioreactor Cultures

  • Song, Sung-Ki;Jeong, Yong-Seob;Kim, Pyeung-Hyeun;Chun, Gie-Taek
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1690-1698
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    • 2006
  • In order to investigate the effect of dissolved oxygen (DO) level on AVM $B_{1a}$ production by a high yielding mutant of Streptomyces avermitilis, five sets of bioreactor cultures were performed under variously controlled DO levels. Using an online computer control system, the agitation speed and aeration rate were automatically controlled in an adaptive manner, responding timely to the oxygen requirement of the producer microorganism. In the two cultures of DO limitation, the onset of AVM $B_{1a}$ biosynthesis was observed to casually coincide with the fermentation time when oxygen-limited conditions were overcome by the producing microorganism. In contrast, this phenomenon did not occur in the parallel fermentations with DO levels controlled at around 30% and 40% throughout the entire fermentation period, showing an almost growth-associated mode of AVM $B_{1a}$ production: AVM $B_{1a}$ biosynthesis under the environments of high DO levels started much earlier than the corresponding oxygen-limited cultures, leading to a significant enhancement of AVM $B_{1a}$ production during the exponential stage. Consequently, approximately 6-fold and 9-fold increases in the final AVM $B_{1a}$ production were obtained in 30% and 40% DO-controlled fermentations, respectively, especially when compared with the culture of severe DO limitation (the culture with 0% DO level during the exponential phase). The production yield ($Y_{p/x}$), volumetric production rate (Qp), and specific production rate (${\bar{q}}_p$) of the 40% DO-controlled culture were observed to be 14%, 15%, and 15% higher, respectively, than those of the parallel cultures that were performed under an excessive agitation speed (350 rpm) and aeration rate (1 vvm) to maintain sufficiently high DO levels throughout the entire fermentation period. These results suggest that high shear damage of the high-yielding strain due to an excessive agitation speed is the primary reason for the reduction of the AVM $B_{1a}$ biosynthetic capability of the producer. As for the cell growth, exponential growth patterns during the initial 3 days were observed in the fermentations of sufficient DO levels, whereas almost linear patterns of cell growth were observed in the other two cultures of DO limitation during the identical period, resulting in apparently lower amounts of DCW. These results led us to conclude that maintenance of optimum DO levels, but not too high to cause potential shear damage on the producer, was crucial not only for the cell growth, but also for the enhanced production of AVM $B_{1a}$ by the filamentous mycelial cells of Streptomyces avermitilis.