• Title/Summary/Keyword: DCW

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Reduction of Hexavalent Chromium by Escherichia coli ATCC 33456 in Batch and Continuous Cultures

  • Bae, Woo-Chul;Kang, Tae-Gu;Kang, In-Kyong;Won, You-Jung;Jeong, Byeong-Chul
    • Journal of Microbiology
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    • v.38 no.1
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    • pp.36-39
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    • 2000
  • Toxic hexavalent chromium, Cr(VI), was reduced to a less toxic trivalent chromium form by E. coli ATCC 33456. The suitable electron donor for Cr(VI) reduction was glucose. E. coli ATCC 33456 was more resistant to metal cations than other reported Cr(VI) reducing microorganisms. Cell growth was inhibited by the presence of Cr(VI) in a liquid medium and Cr(VI) reduction accompanied cell growth. With a hydraulic retention time of 20 h, Cr(VI) reducing efficiency was 100% to 84% when Cr(VI) concentration in the influent was in the range of 10 to 40 mg L$\^$-1/. Specific rate of Cr(VI) reduction was 2.41 mg Cr(VI) g DCW$\^$-1/ h$\^$-1/ when 40 mg L$\^$-1/ of Cr(VI) influent was used. This result suggested the potential application of E. coli ATCC 33456 for the detoxification of Cr(VI) in Cr(VI) contaminated wastewater.

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Influence of Growth Conditions on Plasmid DNA Production

  • Silva, Filomena;Passarinha, Luis;Sousa, Fani;Queiroz, Joao A.;Domingues, Fernanda C.
    • Journal of Microbiology and Biotechnology
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    • v.19 no.11
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    • pp.1408-1414
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    • 2009
  • The obtention of high yields of purified plasmid DNA is viewed as an essential issue to be considered towards efficient production of DNA vaccines and therapeutic plasmids. In this work, Escherichia coli $DH5\alpha$. bearing the pVAXI-LacZ plasmid was grown in a developed semi-defined medium at different temperatures and tryptone concentrations. Analysis of pDNA yields and E. coli morphology revealed that at higher temperatures (37 and $40^{\circ}C$), higher specific yields and E. coli filamentation were obtained. However, the best results were achieved when a lower tryptone concentration was used. This approach was shown to be a powerful tool to promote plasmid amplification, keeping the desirable plasmid structure, and favoring the attainment of quality. Our results suggest that by using tryptone alone as an amino acid source, pDNA amplification was improved and a specific yield of 20.43 mg pDNA/g dcw was achieved, proving that this strategy can improve pDNA yield even at a small scale.

Cell Recycled Culture of Succinic Acid-Producing Anaerobiospirillum succiniciproducens Using an Internal Membrane Filtration System

  • Lee, Pyung-Cheon;Lee, Sang-Yup;Chang, Ho-Nam
    • Journal of Microbiology and Biotechnology
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    • v.18 no.7
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    • pp.1252-1256
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    • 2008
  • Cell recycled culture of succinic acid-producing Anaerobiospirillum succiniciproducens was anaerobically carried out using an internal membrane filter module in order to examine the physiological response of A. succiniciproducens to a high-cell-density environment. The optimal growth of A. succiniciproducens and its enhanced succinic acid productivity were observed under $CO_2$-rich conditions, established by adding $NaHCO_3$ and $Na_2CO_3$, in the cell recycled system. A. succiniciproducens grew up to 6.50 g-DCW/l, the highest cell concentration obtained so far, in cell recycled cultures. The cells did not change their morphology, which is known to be easily changed in unfavorable or stress environments. The maximum productivity of succinic acid was about 3.3 g/l/h, which is 3.3 times higher than those obtained in batch cultures. These results can serve as a guide for designing highly efficient cell recycled systems for succinic acid at a commercial level.

Improvement of Unsaturated Fatty Acid Production from Porphyridium cruentum Using a Two-Phase Culture System in a Photobioreactor with Light-Emitting Diodes (LEDs)

  • Kim, So Hee;Lee, Ui Hun;Lee, Sang Baek;Jeong, Gwi-Taek;Kim, Sung-Koo
    • Journal of Microbiology and Biotechnology
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    • v.31 no.3
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    • pp.456-463
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    • 2021
  • In this study, the culture conditions for Porphyridium cruentum were optimized to obtain the maximum biomass and lipid productions. The eicosapentaenoic acid content was increased by pH optimization. P. cruentum was cultured with modified F/2 medium in 14-L photobioreactors using a two-phase culture system, in which the green (520 nm) and red (625 nm) light-emitting diodes (LEDs) were used during the first and second phases for biomass production and lipid production, respectively. Various parameters, including aeration rate, light intensity, photoperiod, and pH were optimized. The maximum biomass concentration of 0.91 g dcw/l was obtained with an aeration rate of 0.75 vvm, a light intensity of 300 μmol m-2s-1, and a photoperiod of 24:0 h. The maximum lipid production of 51.8% (w/w) was obtained with a light intensity of 400 μmol m-2s-1 and a photoperiod of 18:6 h. Additionally, the eicosapentaenoic acid and unsaturated fatty acid contents reached 30.6% to 56.2% at pH 6.0.

Asymmetric resolution of racemic styrene oxide using recombinant Escherichia coli harboring epoxide hydrolase of Rhodotorula glutinis (Rhodotorula glutinis 유래의 고효율 재조합 Epoxide Hydrolase를 이용한 라세믹 Styrene Oxide의 비대칭 광학분할)

  • Park, Kyu-Deok;Choi, Sung-Hee;Kim, Hee-Sook;Lee, Eun-Yeol
    • KSBB Journal
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    • v.23 no.5
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    • pp.369-374
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    • 2008
  • The effects of reaction temperature and the addition of various detergents on the enantioselective hyrolysis activity of the recombinant Escherichia coli containing the epoxide hydrolase (EH) gene of Rhodotorula glutinis were investigated for the production of enantiopure styrene oxide. The recombinant E. coli harboring the EH gene from R. glutinis exhibited the enantiopreference toward (R)-styrene oxide with the maximum hydrolytic activity of $165.04{\mu}mol/min/mg$ of dry cell weight (dcw). The addition of 0.5% (w/v) Tween 20 at $10^{\circ}C$ increased the initial hydrolysis rate and enantioselectivity by 1.45-fold and 2.0-fold, respectively. Enantiopure (S)-styrene oxide was prepared with 99% ee enantiopurity and 46.0% yield (theoretical yield=50%) from 20 mM racemic styrene oxide.

Production of 10-deacetylbaccatin III in Taxus cuspidata Suspension Cell Culture (주목 현탁세포배양을 이용한 10-deacetylbaccatin III 생산)

  • Lee, Gue-Wha;Kim, Dong-Il
    • KSBB Journal
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    • v.14 no.6
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    • pp.665-671
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    • 1999
  • In this study, enhanced production of 10-deacetylbaccatin III(10-DAB), a precursor of taxol in semisynthesis, was investigated in Taxus cuspidata cell suspension cultures. The effects of initial inoculum size and sugar concentration were examined to prove the relationship between the production of 10-DAB and cell growth. The cell growth was found to be stimulated in Schenk and Hildebrandt(SH) medium. The lower the inoculum size as well as initial sugar concentration, the faster the cell growth rate. When the initial sugar concentration was dept low, the production of 10-DAB into medium was increased. By using perfusion culture, continuous cell growth was possible until the end of culture and more than 34.67 g/L of cell concentration could by obtained. This is about 2.5 times higher level than that of control batch culture.

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Production of Polyhydroxybutyrate from Crude Glycerol and Spent Coffee Grounds Extract by Bacillus cereus Isolated from Sewage Treatment Plant

  • Lee, Gi Na;Choi, So Young;Na, Jonguk;Youn, HaJin;Jang, Yu-Sin
    • KSBB Journal
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    • v.29 no.6
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    • pp.399-404
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    • 2014
  • Production of biodegradable polymer polyhydroxyalkanoates (PHAs) from industrial wastes exhibits several advantages such as recycle of waste and the production of high valuable products. To this end, this study aimed at isolating from the sewage treatment plant a PHA producing bacterium capable of utilizing wastes generated from biodiesel and food industries. A Bacillus cereus strain capable of producing poly(3-hydroxybutyrate) [P(3HB)] was isolated, which was followed by confirmation of P(3HB) accumulation by gas-chromatographic analyses. Then, the effects of nutrient limitation on P(3HB) production by B. cereus was first examined. Cells cultured in a minimal medium under the limitation of nitrogen, potassium and sulfur suggested that nitrogen limitation allows the highest P(3HB) accumulation. Next, production of P(3HB) was examined from both waste of biodiesel production (crude glycerol) and waste from food industry (spent coffee grounds). Cells cultured in nitrogen-limited minimal medium supplemented crude glycerol and waste spent coffee grounds extract accumulated P(3HB) to the contents of 2.4% and 1.0% of DCW. This is the first report demonstrating the capability of B. cereus to produce P(3HB) from waste raw materials such as crude glycerol and spent coffee grounds.

Production of Glutathione by the Yeast Mutant Saccharomyces cerevisiae Sa59 (효모변이주 Saccharomyces cerevisiae Sa59에 의한 glutathione 생성)

  • Jang, Hye-Yoon;Oh, Chul-Hwan;Oh, Nam-Soon
    • Korean Journal of Food Science and Technology
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    • v.45 no.6
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    • pp.801-804
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    • 2013
  • The glutathione contents of the selected mutants were investigated and found to be 6.1-15.8 mg/g-DCW. The glutathione content positively correlated with the antioxidant activity of the mutant strains ($R^2$=0.488). Furthermore, the glutathione content of the mutant S. cerevisiae Sa-59 was approximately 38% greater than that of the wild type strain and, therefore, this mutant strain was selected for glutathione production. The volumetric glutathione content in a shaking culture was increased by about 70% compared to the static culture. In addition, the specific glutathione content was increased by ~19%. The volumetric glutathione content and specific glutathione content were increased by approximately 16% and 66%, respectively, when 0.04% glutamate, 0.04% cysteine and 0.04% glycine were added. Furthermore, the highest antioxidant activity was 0.52 as absorbance unit at 700 nm.

A Fermentation Strategy for Anti-MUC1 C595 Diabody Expression in Recombinant Escherichia Coli

  • Lan, John Chi-Wei;Ling, Tau Chuan;Hamilton, Grant;Lyddiatt, Andrew
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.5
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    • pp.425-431
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    • 2006
  • The development of fermentation conditions for the production of C595 diabody fragment (dbFv) in E. coli HB2151 clone has been explored. Investigations were carried out to study the effect of carbon supplements over the expression period, the comparison of C595 dbfv production in synthetic and complex media, the influence of acetic acid upon antibody production, and comparison of one-stage and two-stage processes operated at batch or fed-batch modes in bioreactor. Yeast extract supplied during expression yielded more antibody fragment than any other carbon supply. The synthetic medium presented higher specific productivity (0.066 mg dbFv $g^{-1}$ dry cell weight) when compared to the complex medium (0.044 mg dbFv $g^{-1}$ DCW). The comparison of fermentation strategies demonstrated that (1) one-stage fed-batch fermentation performed higher C595 dbFv production than that operated in batch mode which was significantly affected by acetate concentration; (2) a two-stage batch operation could enhance C595 dbFv production. It was found that a concentration of 12.3 mg $L^{-1}$ broth of C595 dbFv and a cell concentration of 10.8g $L^{-1}$ broth were achieved at the end of two-stage operation in 5-L fermentation.

Enantioselective Kinetic Resolution of Racemic Styrene Oxide using Recombinant Marine Fish Epoxide Hydrolase of Mugil cephalus (해양 어류 Mugil cephalus 유래의 에폭사이드 가수분해효소를 이용한 라세믹 styrene oxide의 입체선택적 분할 반응)

  • Choi, Sung Hee;Kim, Hee Sook;Lee, Eun Yeol
    • Applied Chemistry for Engineering
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    • v.19 no.5
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    • pp.491-496
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    • 2008
  • The microsomal epoxide hydrolase gene (referred to as mMCEH) of Mugil cephalus was cloned by PCR, and then inserted to pColdI and pET-21b(+) vector, respectively. The recombinant E. coli possessing the recombinant plasmids exhibited the enantioperference toward (R)-styrene oxide. When enantioselective kinetic resolutions were conducted with 20 mM racemic styrene oxide, enantiopure (S)-styrene oxide was obtained with high enantiopurity more than 99% enantiomeric excess (ee) and 24.50% yield by using the recombinant E. coli harboring pET-21b(+)/mMCEH.