• Title/Summary/Keyword: Medium Optimization

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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.

Optimization of Fermentation Conditions for Production of Recombinant Human Interleukin-2 in Escherichia coli (대장균에서의 재조합 인체 인터루킨-2 생산을 위한 발효조건 최적화)

  • Lee, In-Young;Kim, Myung-Kuk;Na, Doe-Sun;Hahm, Kyung-Soo;Moon H. Han;Lee, Sun-Bok
    • Microbiology and Biotechnology Letters
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    • v.16 no.4
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    • pp.327-333
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    • 1988
  • For optimal production of recombinant human interleukin-2 (IL-2) in E. coli the effect of fermentation conditions on cell growth, IL-2 production, and stability of recombinant cells were investigated. Among the complex nutrients tested in this work, yeast extract, peptone and corn steep liquor were found to be effective for recombinant cell growth. The recombinant cells were maintained stably under repression condition (3$0^{\circ}C$), but the stability of recombinant cells were drastically reduced upon induction of IL-2 expression (42$^{\circ}C$) even under the selection pressure. Addition of antibiotics to the culture medium resulted in the cell growth inhibition without significant improvement in recombinant stability. When the expression of IL-2 gene was induced at different growth phases, highest IL-2 production was achieved by the induction of IL-2 at the middle-exponential growth phase. It was found that the production of IL-2 significantly inhibited the cell growth and the ex-pression of other genes in the plasmid.

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Mass Production of Pullulan with Optimized Concentrations of Carbon and Nitrogen Sources by Aureobasidium pullulans HP-2001 in a 100-L Bioreactor with the Inner Pressure

  • Seo, Hyung-Pil;Chung, Chung-Han;Kim, Sung-Koo;Richard A. Gross;David L. Kaplan;Lee, Jin-Woo
    • Journal of Microbiology and Biotechnology
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    • v.14 no.2
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    • pp.237-242
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    • 2004
  • Cell growth and the production of pullulan by Aureobasidium pullulan HP-2001, the UV-induced mutant of A pullulans ATCC 42023, increased with increased concentration of glucose up to 15.0% (w/v). Maximal production of pullulan in the flask scale was 27.65 g/l, when concentrations of glucose and yeast extract were 15.0 and 0.25% (w/v), respectively. Maximal conversion rate of pullulan from glucose as the carbon source was 0.37, when those of glucose and yeast extract were 5.0 and 0.15% (w/v), respectively. On the basis of total amount of pullulan, the conversion rate of pullulan from glucose, and utilization rate of glucose to cell mass and pullulan by A. pullulans HP-2001, the optimal concentrations of glucose and yeast extract for the mass production of pullulan were determined to be 10.0 and 0.25% (w/v), respectively, at which concentrations the production of pullulan and its conversion rate were 27.14 g/l and 0.27, respectively. Maximal production of pullulan with optimized concentrations of carbon and nitrogen sources by A. pullulans HP-200l in a 7-1 bioreactor was 32.12 g/l for 72 h culture, and that in a 100-1 bioreactor with the inner pressure of $0.4 kgf/cm^2$ was 36.87 g/l. Increased inner pressure of a 100-1 bioreactor resulted in a higher concentration of dissolved oxygen in the medium, which might enhance the production of pullulan by A. pullulans HP-2001.

Characteristics of Culture Conditions for the Production of Crude Biosurfactant by Bacillus subtilis JK-1 (Bacillus subtilis JK-1의 생물계면활성제 생산을 위한 배양 특성)

  • Kim, Ji-Yeon
    • Journal of Applied Biological Chemistry
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    • v.54 no.3
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    • pp.153-158
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    • 2011
  • Optimal culture conditions were characterized for production of crude biosurfactant of Bacillus subtilis JK-1. During incubation of B. subtilis JK-1, the bacterial growth pattern, changes of the surface tension at variable temperatures, pH and NaCl concentrations in bacterial culture medium were studied. The strain was able to grow and produce biosurfactant at $15-45^{\circ}C$, in the pH range of 6-10, and at 0-10% (w/v) NaCl. In case, culture broth pH was gradually changed to neutral or weak alkaline. Optimal culture conditions for crude biosurfactant production were at $35^{\circ}C$ and pH 7.0 after 48 h incubation and the surface tension of biosurfactant was 24.0 mN/m. Besides, as the concentration of NaCl was increased from 0 to 10% (w/v), the growth was decreased, pH of the culture broth was converted from weak alkaline to acidic, and the surface tension rised.

Feasibility Study of High Strength Steel on Steel Bridge (고강도 강재의 강교량 적용성에 관한 연구)

  • Jeon, Jun Chang;Kim, Seok Tae;Kyung, Kab Soo;Lee, Hee Hyun
    • Journal of Korean Society of Steel Construction
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    • v.14 no.5 s.60
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    • pp.603-612
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    • 2002
  • Numerical analyses have been carried out in order to check the applicability of high-strength steel to a medium-sized steel bridge. Using the yield strength of steel, Average Daily Truck Traffic (ADTT), and fatigue grade of structural detail as analytical parameters, the minimum weight sections that satisfy the limit states of the AASHTO LRFD design specification were determined through an optimization scheme. Likewise, the effects of the number of girders and span length on the applicability of high-strength steel were evaluated. Results show that high-strength steel may be employed in the steel bridge, since steel weight decreases with increasing yield strength regardless of the fatigue effect. Nonetheless, appropriate countermeasures against fatigue should be determined since it is a major factor in the effective use of high-strength steel in steel bridges.

Effect of Light-Emitting Diodes on Cordycepin Production in Submerged Culture of Paecilomyces japonica

  • HA, Si Young;JUNG, Ji Young;YANG, Jae-Kyung
    • Journal of the Korean Wood Science and Technology
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    • v.48 no.4
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    • pp.548-561
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    • 2020
  • Paecilomyces japonica is widely cultured to produce mycelium for medicinal and health food use. Illumination is an important factor in the growth and production of mycelium in submerged culture. The effects of different light-emitting diode (LED) combinations on the growth and cordycepin content as bioactive substances of mycelium were investigated. The results showed that the mycelium dry weights were lower under dark condition and red LED treatments. Dark condition, fluorescent light, and ultraviolet-A failed to increase the cordycepin content. Blue light was necessary to increase the cordycepin content, and a red-to-blue ratio of 3:7 induced the highest cordycepin content. The cordycepin contents of mycelium in submerged culture were significantly higher in a 12 h/day illumination time under red and blue (red-to-blue ratio of 3:7) LED treatments, showing an increase of up to 38% compared with those under the fluorescent-light control condition. The results demonstrated the roles of light with different wavelengths on the biosynthesis of cordycepin as bioactive substances. The low-heat release and replacement of traditional fluorescent lights with low-energy-consuming LEDs could increase the contents of bioactive substances. After optimization of the cordycepin production using response surface methodology (Box-Behnken design) to its canonical form, the optimum combination was found to be as follows: illumination time = 17.7 h/day, sugar content in the medium = 9.7 g/50 mL, and incubation time = 61.2 h. The model predicted a maximum response of 3779.2 ㎍/mL cordycepin yield.

Optimization of bioactive isorhamnetin 3-O-glucoside production in Escherichia coli (대장균에서 isorhamnetin 3-O-glucoside의 생합성 최적화)

  • Kim, Bong-Gyu
    • Journal of Applied Biological Chemistry
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    • v.62 no.4
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    • pp.361-366
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    • 2019
  • Isorhamnetin 3-O-glucoside, a member of the flavonol group, has been reported to be effective for inflammatory and ulcer, as well as to alleviate diabetic complications such as neuropathy, nephropathy and retinopathy. Isorhamnetin 3-O-glucoside has been extracted from several plants. Biotransformation is a valuable tool, which is used to produce value-added chemicals with inexpensive compounds. To synthesis isorhamnetin 3-O-glucoside from quercetin, two genes (PGT E82L and ROMT-9) were introduced into Escherichia coli, respectively. In order to synthesis isorhamnetin 3-O-glucoside from quercetin, a co-culture fermentation system was developed by optimizing the medium and temperature for biotransformation, the cell mix ratio, Isopropyl-β-ᴅ-thiogalactoside induction time, and quercetin feed concentration. Finally, isorhamnetin 3-O-glucoside was biosynthesized up to 181.2 mg/L under the optimized biotransformation condition, which was higher 4.7 times than previously reported (39.6 mg/L).

The Secretion Optimization of Oligopeptide with His-Pro Repeats in Bacillus subtilis and Its Anti-Diabetic Effects (고초균에서 His-Pro 반복서열을 갖는 Oligopeptide의 분비 최적화 및 항당뇨 효과)

  • Jeong, Seon Hwa;Choi, Jang Won
    • KSBB Journal
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    • v.32 no.1
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    • pp.71-82
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    • 2017
  • To verify anti-diabetic effect of oligopeptide with His-Pro repeats (mHP peptide), the oligopeptide was first secreted and optimized using the secretion vector, pRBAS with alkaline protease gene promoter and the signal sequence in Bacillus subtilis and directly the anti-diabetic effect of the mHP peptide was investigated in insulinoma cell, RINm5F cell line. The oligopeptide gene was obtained by annealing oligonucleotides with repeated His-Pro sequence and finally was constructed as 18 dipeptides (108 bp and 4.0 kDa) coding gene, named oligopeptide with His-Pro repeats (mHP peptide) to make cyclo(His-Pro) known to be anti-diabetic effects. The region encoding the oligopeptide gene was subcloned into the pRBAS secretion vector (E.coli-Bacillus shuttle vector) after PCR amplification using the designed primers including initiation and termination codons and His tag, named pRBAS-mHP (6.56 kb). To optimize secretion of the oligopeptide, various culture conditions were investigated in Bacillus subtilis LKS. As a result, the secreted oligopeptide was maximally measured (approximately $59.6{\mu}g/mL$) in 3 L batch culture and the highest secretion was achieved at $30^{\circ}C$, PY medium, and carbon sources (particularly barley and glycerol). In the RINm5F cells treated with 2 mM STZ, the oligopeptide treatment (0.1 mg/mL) restored the cell viability (10%) and reduced the nitric oxide (NO) generation (35%) and DNA fragmentation (90%). And also, insulin secretion level was increased to 17% higher than in STZ-treated RINm5F cells. These results suggest that the oligopeptide with His-Pro repeats could be a candidate material for anti-diabetic agent against STZ-induced diabetes.

Biological characteristics of Paenibacillus polymyxa GBR-1 involved in root rot of stored Korean ginseng

  • Kim, Young Soo;Kotnala, Balaraju;Kim, Young Ho;Jeon, Yongho
    • Journal of Ginseng Research
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    • v.40 no.4
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    • pp.453-461
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    • 2016
  • Background: This study aims to describe the characterization of Paenibacillus polymyxa GBR-1 (GBR-1) with respect to its positive and negative effects on plants. Methods: The morphological characteristics of GBR-1 were identified with microscopy, and subjected to Biolog analysis for identification. Bacterial population and media optimization were determined by a growth curve. The potential for GBR-1 as a growth promoting agent, to have antagonistic activity, and to have hydrolytic activity at different temperatures was assessed. The coinoculation of GBR-1 with other microorganisms and its pathogenicity on various stored plants, including ginseng, were assessed. Results: Colony morphology, endospore-bearing cells, and cell division of GBR-1 were identified by microscopy; identification was performed by utilizing the Biolog system, gas chromatography of fatty acid methyl esters (GC-FAME). GBR-1 showed the strongest antagonistic activity against fungal and bacterial pathogens. GBR-1 cell numbers were relatively higher when the cells were cultured in brain heart infusion (BHI) medium when compared with other media. Furthermore, the starch-hydrolytic activity was influenced by GBR-1 at higher temperature compared to low temperatures. GBR-1 was pathogenic to some of the storage plants. Coinoculation of GBR-1 with other pathogens causes differences in rotting on ginseng roots. A significant growth promotion was observed in tobacco seedlings treated with GBR-1 suspensions under in vitro conditions, suggesting that its volatile organic compounds (VOCs) might play a role in growth promotion. Conclusion: The results of this study indicate that GBR-1 has both positive and negative effects on ginseng root and other stored plants as a potential biocontrol agent and eliciting in vitro growth promotion.

Optimization of Anticoagulant Production from Phellinus linteus Mycelia (상황버섯(Phellinus linteus) 균사체로부터 항혈전 생산의 최적화)

  • Seo, Ho-Chan
    • The Korean Journal of Mycology
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    • v.39 no.2
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    • pp.117-121
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    • 2011
  • To produce the functional food materials, 50 kinds of the mycelial extracts from edible mushroom were examined for anticoagulant activity and Phellinus linteus showed the highest activity through the activated partial thromboplastin test (aPTT). The maximum production of anticoagulant activity and the mycelial growth was observed in culture medium containing soluble starch 3.0%, peptone 0.1%, $MgSO_4{\cdot}7H_2O^{\circ}C$ 0.1%, $K_2HPO_4$ 0.1% and in the culture conditions controlled at initial pH 7.0, $30^{\circ}C$ and 150 rpm by the rotary shaker. In addition, the maximum production of mycelial dry weight was 7.5 mg/mL after 10 days under the optimal conditions, and anticoagulant activity was reached to 390 sec in 5 L-jar fermentor.