• Title/Summary/Keyword: orbital shaker

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해양 곰팡이 Thraustochytrium aureum의 형태학적 변화와 DHA 생산과의 특성 규명

  • Park, Gyeong-Won;Park, Cheon-Ik;Jo, Dae-Won;Heo, Byeong-Gi
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.273-276
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    • 2000
  • Marine fungus, Thraustochytrium aureum ATCC 34034, was incubated in artificial sea water media in order to produce docosahexaenoic acid(DHA). Cultures were performed at $24^{\circ}C$ in light for 13 days with orbital shaker at 100rpm. Maximum cell productivity of 1.34g/L and DHA yield of 41.4 mg/L were obtained by using this method, which is almost the twice level of DHA yield obtained for the strains reported previously. This strain did produce much more DHA after sporangium disruption.

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Combined TGE-SGE Expression of Novel PAI-1-Resistant t-PA in CHO DG44 Cells Using Orbitally Shaking Disposable Bioreactors

  • Davami, Fatemeh;Barkhordari, Farzaneh;Alebouyeh, Mahmoud;Adeli, Ahmad;Mahboudi, Fereidoun
    • Journal of Microbiology and Biotechnology
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    • v.21 no.12
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    • pp.1299-1305
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    • 2011
  • An important modification of thrombolytic agents is resistance to plasminogen activator inhibitor-1 (PAI-1). In previous studies, a new truncated PAI-1-resistant variant was developed based on deletion of the first three domains in t-PA and the substitution of KHRR 128-131 amino acids with AAAA in the truncated t-PA. The novel variant expressed in a static culture system of Chinese Hamster Ovary (CHO) DG44 cells exhibited a higher resistance to PAI-1 when compared with the full-length commercial drug; Actylase. In the present study, the truncated-mutant protein was expressed in CHO DG44 cells in 50 ml orbital shaking bioreactors. The final yield of the truncated-mutant in the culture was 752 IU/ml, representing a 63% increase compared with the static culture system. Therefore, these results suggest that using the combined features of a transient and stable expression system is feasible for the production of novel recombinant proteins in the quantities needed for preclinical studies.

고수율 DHA생산을 위한 Thraustochytrium aureum의 배지조성과 발효조건에 관한 연구

  • Park, Gyeong-Won;Kim, Jun-Sik;Heo, Byeong-Gi
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.291-294
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    • 2000
  • Marine fungus, Thraustochytrium aureum ATCC 34034, was incubated in artificial sea water media in order to produce docosahexaenoic acid(DHA). Cultures were performed at $24^{\circ}C$ in light for 3 days with orbital shaker at 200rpm. Maximum cell productivity of 1.34g/L and DHA yield of 41.4 mg/L were obtained by using this method, which is almost the twice level of DHA yield obtained for the strains reported previously. This strain did not produce DHA in YM media and YPG media. The results indicate that salinity influenced cell growth and production of DHA.

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Citric Acid Production from Glucose and Pumpkin by Using Immobilized Bead of Aspergillus niger (고정화 Aspergillus niger Bead를 이용한 포도당과 호박으로부터 구연산 생성)

  • 방병호
    • The Korean Journal of Food And Nutrition
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    • v.13 no.4
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    • pp.328-333
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    • 2000
  • The spore of Aspergillus niger KCTC-6144 was immobilized on alginate gel beads. When pumpkin powder was used with glucose for a medium of citric acid fermentation by Aspergillus niger beads, the beaded Aspergilus niger grew up inside the bead and mycelia penetrated through the pore of the bead membrane. The bead size became largely from 2.0∼2.5mm to 6∼8mm after growing at 30$\^{C}$ for 4 days. Studies of optimum culture conditions on citric acid fermentation using Aspergillus niger beads on pumpkin medium (pumpkin powder 1% +glucose 7%, pH 6.0) were carried out in submerged cultures on 250m1 Erlenmeyer flask. As a result, it was found that to reinforce 12% as carbon source was good for citric acid production and that 1% pumpkin powder was good as nitrogen and mineral source in orbital shaker (150rpm) at 30$\^{C}$ for 5 days. The optimum initial pH on citric acid production was pH 6.0 and it was found that 100 beads of immobilized Aspergillus niger was adequate for citric acid production in a 250ml Erlenmeyer flask containing 50m3 of pumpkin medium solution with orbital shaker at 30$\^{C}$ for 5 days. We also found that maximal production of citric acid was 23.5g/ℓ at optimal condition (at 30$\^{C}$ for 5 days, pH 6.0, and 100 beads and medium containing 1% pumpkin powder plus 12% glucose).

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Production of Indole-3-Acetic Acid by Enterobacter sp. DMKU-RP206 Using Sweet Whey as a Low-Cost Feed Stock

  • Srisuk, Nantana;Sakpuntoon, Varunya;Nutaratat, Pumin
    • Journal of Microbiology and Biotechnology
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    • v.28 no.9
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    • pp.1511-1516
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    • 2018
  • In this study, we investigated Indole-3-acetic acid (IAA) production by a rice phylloplane bacteria, Enterobacter sp. DMKU-RP206, using sweet whey as a feed stock instead of lactose. We succeeded in using sweet whey for Enterobacter sp. DMKU-RP206 to produce 3,963.0 mg IAA/l with the optimal medium containing 1.48% sweet whey, 1.42% yeast extract and 0.88% $\text\tiny{L}$-tryptophan. The medium pH was adjusted to 6 and the culture conditions were shaking at 200 rpm on an orbital shaker at $30^{\circ}C$ for 3 days. We also evaluated the effect of IAA in culture filtrates of Enterobacter sp. DMKU-RP206 on the promotion of jasmine rice growth in a pot experiment. Compared with the negative control (without IAA), the result showed that biosynthetic IAA produced by Enterobacter sp. DMKU-RP206 significantly increased the growth of jasmine rice (Oryza sativa L. cv. KDML105) in terms of length and dry weight of shoot. This work thus reveals the impact of IAA produced by Enterobacter sp. on the promotion of jasmine rice growth.

Enhanced pectinase and β-glucosidase enzyme production by a Bacillus subtilis strain under blue light-emitting diodes

  • Elumalai, Punniyakotti;Lim, Jeong-Muk;Oh, Byung-Teak
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.10a
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    • pp.109-109
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    • 2018
  • Bacillus subtilis B22, a chemotrophic and aerobic bacterial strain was isolated from homemade kimchi, identified by 16S rRNA gene sequencing. B22 was primarily screened by biochemical, carbon source utilization tests. B22 was used to produce pectinase and ${\beta}$-glucosidase by submerged fermentation under different light sources. B22 was incubated in pectin media and basal media (pH 7.0) under blue, green, red and white light-emitting diodes (LEDs), fluorescent white light, and in darkness at $37^{\circ}C$, orbital shaker 150 rpm for 24 hours. Fermentation under blue LEDs maximized pectinase production ($71.59{\pm}1.6U/mL$ at 24 h) and ${\beta}$-glucosidase production ($56.31{\pm}1.6U/mL$ at 24 h). Further, the production of enzyme increased to pectinase ($156{\pm}1.28U/mL$) and ${\beta}$-glucosidase ($172{\pm}1.28U/mL$) with 3% glucose as a carbon source. Activity and stability of the partially purified enzymes were higher at pH 6.0 to 8.0 and $25-55^{\circ}C$. The effect on the metal ions $Na^+$ and $K^+$ and (moderateactivity) $Mn^{2+}$ and $Ni^{2+}$ increased activity, while $Hg^{2+}$, $Cu^{2+}$, $Fe^{2+}$, and $Fe^{2+}$ inhibited activity. EDTA, phenylmethylsulfonyl fluoride and 5,5-dithiobis (2-nitrobenzoicacid) reduced activity, while tetrafluoroethylene and 1,10-phenanthroline inhibited activity. The amylase was highly tolerant of the surfactants TritonX-100, Tween-20, Tween-80 and compatible with organic solvents methanol, ethanol, isoamylalcohol, isopropanol, t-butylalcohol and the oxidizing agents hydrogen peroxide, sodium perborate and sodium hypochlorite, although potassium iodide and ammonium persulfate reduced activity. These properties suggest utility of pectinase and ${\beta}$-glucosidase produced by B. subtilis B22 under blue LED-mediated fermentation for industrial applications.

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Immune Response of Bacterial Proteins of Staphylococcus intermedius from Canine Atopic Dermatitis (개의 아토피성 피부염에서 분리한 Staphylococcus intermedius 균의 세균단백질의 면역반응)

  • Park, Hee-myung
    • Journal of Veterinary Clinics
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    • v.21 no.1
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    • pp.20-22
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    • 2004
  • Bacterial infection of canine atopic dermatitis is largely caused by Staphylococcus intermedius and may be a superficial or deep pyoderma. The Purpose of this study was to identify the major proteins of S. intermedius cell surface components in humoral immune response of atopic dermatitis dog. Sera samples were obtained from dogs with atopic dermatitis and superficial pyoderma referred to the Veterinary Medical Teaching Hospital, Konkuk University. An isolate of S. intermedius from a clinical case of canine atopic dermatitis was cultured in brain heart infusion broth overnight at $37^{\circ}C$ in aerobic conditions on an orbital shaker. Following culture, Staphylococci were harvested by centrifugation, washed in PBS, and resuspended in PBS containing lysostaphin. The soluble components were separated by centrifugation and were collected. The soluble extract of S. intermedius was separated by sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE). The proteins were electrophoretically transferred onto nitrocellulose membrane. Western blotting for the specificity of serum IgG antistaphylococcal antibody was performed with anti-dog-IgG and sera obtained from an atopic dermatitis case and a normal dog. The molecular masses of four major proteins of S. intermedius recognized by serum obtained from an atopic dermatitis case were 18, 31, 75, and 110 kDa as determined by Western blot analysis. The present study indicates that most dogs of S. intermedius infection with atopic dermatitis could have a significant humoral immune response to bacterial proteins of the causative organism.