• Title/Summary/Keyword: whole cell conversion

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Cofactor Regeneration Using Permeabilized Escherichia coli Expressing NAD(P)+-Dependent Glycerol-3-Phosphate Dehydrogenase

  • Rho, Ho Sik;Choi, Kyungoh
    • Journal of Microbiology and Biotechnology
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    • v.28 no.8
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    • pp.1346-1351
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    • 2018
  • Oxidoreductases are effective biocatalysts, but their practical use is limited by the need for large quantities of NAD(P)H. In this study, a whole-cell biocatalyst for NAD(P)H cofactor regeneration was developed using the economical substrate glycerol. This cofactor regeneration system employs permeabilized Escherichia coli cells in which the glpD and gldA genes were deleted and the gpsA gene, which encodes $NAD(P)^+-dependent$ glycerol-3-phosphate dehydrogenase, was overexpressed. These manipulations were applied to block a side reaction (i.e., the conversion of glycerol to dihydroxyacetone) and to switch the glpD-encoding enzyme reaction to a gpsA-encoding enzyme reaction that generates both NADH and NADPH. We demonstrated the performance of the cofactor regeneration system using a lactate dehydrogenase reaction as a coupling reaction model. The developed biocatalyst involves an economical substrate, bifunctional regeneration of NAD(P)H, and simple reaction conditions as well as a stable environment for enzymes, and is thus applicable to a variety of oxidoreductase reactions requiring NAD(P)H regeneration.

Production of propylene oxide from propene by a methanotroph, Methylosinus trichosporium OB3b (Methane 자화성 세균 Methylosinus trichosporium OB3b에 의한 propene으로부터 propylene oxide의 생산)

  • Chung, Dae-Seok;Peck, Un-Hwa;Bang, Wong-Gi
    • Applied Biological Chemistry
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    • v.34 no.4
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    • pp.386-392
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    • 1991
  • Whole cells of Methylosinus trichosporium OB3b, the obligate methylotroph, were used to produce propylene oxide from propane. This strain has methane monooxygenase, which catalyzes the conversion methane to methanol and can catalyze also the conversion propane to propylene oxide. Optimal condition for the production of propylene oxide was investigated in resting-whole cell system. The optimal pH and temperature was 7.0 and $35^{\circ}C$ respectively. The end product, propylene oxide, didn't inhibit the production of propylene oxide and was not further metabolized in reaction mixture. The addition of methane metabolites (methanol, formaldehde and formic acid) to the reaction mixture stimulated formation of propylene oxide by $3{\sim}4$ times, and methanol was the most effective especially. Under the optimal conditions, the 14.2 mM of propylene oxide was produced after incubation of 60 min. and the conversion ratio of propane to propylene oxide was approximately 8%.

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Synthesis and Photovoltaic Performance of Long Wavelength Absorption Dyes for the Dye Sensitized Solar Cell (장 파장 대 태양광을 흡수하는 염료감응형태양전지에 대한 염료와 합성)

  • Kim, Sangah;Yoon, Jooyoung;Kim, Jaehong
    • 한국신재생에너지학회:학술대회논문집
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    • 2010.06a
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    • pp.89.2-89.2
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    • 2010
  • The dye-sensitized solar cell (DSSC) is a device for the conversion of visible light into electricity, based on the sensitization of wide bandgap semiconductors. The performance of the cell mainly depends on a dye used as sensitizer. The absorption spectrum of the dye and the anchorage of the dye to the surface of $TiO_2$ are important parameters determining the efficiency of the cell. Generally, transition metal coordination compounds(ruthenium polypyridyl complexes) are used as the effective sensitizers, due to their intense charge-transfer absorption in the whole visible range and highly efficient metal-to ligand charge transfer. However, ruthenium polypyridyl complexes contain a heavy metal, which is undesirable from point of view of the environmental aspects. Moreover, the process to synthesize the complexes is complicated and costly. Alternatively, organic dyes can be used for the same purpose with an acceptable efficiency. The advantages of organic dyes include their availability and low cost. We designed and synthesized a series of organic sensitizers containing long wavelength absorption-chromophores for the dye sensitized solar cell. The DSSC composed of Blue-chromophores for the sensitization absorbed long wavelength region which is different also applied into the dye-cocktail (mixing) system. The photovoltaic property of DSSCs organic long wavelength absorption-chromophores were measured and evaluated by comparison with that of individual chromophores.

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Synthesis and Photovoltaic Performance of NIR Absorption Dyes for the Dye Sensitized Solar Cell (NIR 흡수 염료를 이용한 염료감응형 태양전지)

  • Kim, Sangah;Jung, Miran;Lee, Minkyung;Kim, Jaehong
    • 한국신재생에너지학회:학술대회논문집
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    • 2011.05a
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    • pp.118.1-118.1
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    • 2011
  • The dye-sensitized solar cell (DSSC) is a device for the conversion of visible light into electricity, based on the sensitization of wide bandgap semiconductors. The performance of the cell mainly depends on a dye used as sensitizer. The absorption spectrum of the dye and the anchorage of the dye to the surface of TiO2 are important parameters determining the efficiency of the cell. Generally, transition metal coordination compounds(ruthenium polypyridyl complexes) are used as the effective sensitizers, due to their intense charge-transfer absorption in the whole visible range and highly efficient metal-to ligand charge transfer. However, ruthenium polypyridyl complexes contain a heavy metal, which is undesirable from point of view of the environmental aspects. Moreover, the process to synthesize the complexes is complicated and costly. Alternatively, organic dyes can be used for the same purpose with an acceptable efficiency. The advantages of organic dyes include their availability and low cost. We designed and synthesized a series of organic sensitizers containing long wavelength absorption-chromophores for the dye sensitized solar cell. The DSSC composed of Blue-chromophores for the sensitization absorbed long wavelength region which is different also applied into the dye-cocktail (mixing) system. The photovoltaic property of DSSCs organic long wavelength absorption-chromophores were measured and evaluated by comparison with that of individual chromophores.

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Effect of the Molar H2O/ and the Molar O2/C Ratio on Long-Term Performance of Diesel Autothermal Reformer for Solid Oxide Fuel Cell (고체산화물 연료전지용 디젤 자열개질기의 장기성능에 미치는 H2O/C와 O2/C 몰 비의 영향)

  • Yoon, Sang-Ho;Kang, In-Yong;Bae, Gyu-Jong;Bae, Joong-Myeon
    • Journal of the Korean Electrochemical Society
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    • v.10 no.2
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    • pp.110-115
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    • 2007
  • Solid oxide fuel cell(SOFC) has high fuel flexibility due to its high operating temperatures. Hydrocarbonaceous fuels such as diesel has several advantages such as high energy density and established infrastructure for fuel cell applications. However diesel reforming has technical problems like coke formation in a reactor, which results in catastrophic failure of whole system. Performance degradation of diesel autothermal reforming (ATR) leads to increase of undesirable hydrocarbons at reformed gases and subsequently degrades SOFC performance. In this study, we investigate the degradation of SOFC performance(OCV, open circuit voltage) under hydrocarbon(n-Butane) feeds and characteristics of diesel performing under various ratios of reactants($H_2O/C,\;O_2/C$ molar ratios) for improvement of SOFC performance. Especially we achieved relatively high performance of diesel ATR under $H_2O/C=0.8,\;O_2/C=3$ condition.

Economic Production of $\gamma$-Interferon from Recombinant Human Cells in Serum Free Medium by a Moving Aeration Membrane Bioreactor (교반형 막 반응기를 이용한 재조합 인간 세포의 무혈청 배지에 의한 $\gamma$-Interferon의 생산)

  • Park, Young-Shik;Kim, Hyun-Kyu;Lim, Seo-Kyu;Park, Kyung-You;Lee, Hyeon-Yong
    • Microbiology and Biotechnology Letters
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    • v.22 no.4
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    • pp.389-394
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    • 1994
  • 8 X 10$^{6}$(viable cells/ml) of maximum cell density and 9000(IU/ml) of $\gamma$-IFN production were obtained at 55(ml/hr) of a perfusion rate by cultivating HSF cells using a moving membrane aeration bioreactor. This system proves to be an efficient culture process by maintaning 90% of viable cells during the whole cultivation periods. The metabolic molar quotient of glucose to lactate was 0.81 for overall ranges of glucose consumed while the evolution of ammonia was not linearly related to the consumption of glutamine. Low molar conversion ratio was observed in low consumptions of glutamine and high molar conversion ratio in high comsumptions. It also shows that the glutamolysis plays important role in the steady state conditions by evolving larger quantities of ammonia than lactate. At the above of 50 rpm, which is the optimum agitation speed for this bioreactor, the cell growth was severely affected while the IFN production was less decrea- sed, maintaing 1.5 X 10$^{-3}$(IU/cell/day) specific IFN production rate. The cumulatvie $\gamma$-IFN production was 7.2 X 10$^{8}$(IU) for 70 days of the cultivation, which yields 1 X 10$^{7}$ (IU/day) of IFN production rate. Therefore, a commercial production of $\gamma$-IFN by this culture process can be achievable by maintaining the above IFN productivity in a scaled-up culture system.

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Evaluation and cloning of a (R)-stereospecific esterase from Bacillus stearothermophilus JY144

  • Kim, Ji-Yeon;Kim, Yun-Jeong;Choe, Gi-Seop;Kim, Geun-Jung;Yu, Yeon-U
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.457-460
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    • 2002
  • In an effort to isolate novel strains expressing a thermostable esterase that hydrolyzed the rac-ketoprofen ethyl ester to ketoprofen in the stereospecific manner, we screened various soils and composts from broad ecological niches in which the activity was expected to be found. Three hundreds of microbial strains were tested to determine their ester-hydrolyzing activity by using an agar plate containing insoluble tributyrin as an indicative substrate, and then further screened by activity on the (R,S)-ketoprofen ethyl ester. Twenty-six strains were screened primarily at high growth and incubation temperature and further compared the ability to ethyl ester-hydrolyzing activity in terms of conversion yield and chiral specificity. Consequently, a strain JYl44 was isolated as a novel strain that produced a (R)-stereospecific esterase with high stability and systematically identified as a Bacillus stearothermophilus JY144. The enzyme indeed stables at a broad range of temperature, upto 65 $^{\circ}C$, and pH ranging from 6.0 to 10.0. The optimal temperature and pH for enzymatic conversion were 50 $^{\circ}C$ and 9.0, respectively. Based on the observations that resulted a poor cell growth, and enzyme expression in wild type strain, we further attempted the gene cloning into a general host Escherichia coli and determined its primary structure, concomitantly resulting a high level expression of the enzyme. The cloned gene had an open reading frame (250 amino acids) with a calculated molecular mass of 27.4 kDa, and its primary structure showed a relative high homology (45-52 %) to the esterases from Streptomyces and Bacillus strains. The recombinant whole cell enzyme could efficiently convert the rac-ketoprofen ethyl ester to (R)-ketoprofen, with optical purity of 99 % and yield of 49 %.

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A New Efficient Mppt Control Algorithm for Low Insolation Intensity

  • Yu, Gwon-Jong;Jung, Young-Seok;Park, Ju-Yeop
    • KIEE International Transaction on Electrical Machinery and Energy Conversion Systems
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    • v.2B no.4
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    • pp.214-218
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    • 2002
  • In this paper, the effectiveness of three different control algorithms are thoroughly investigated via simulation and a proposed efficiency evaluation method of experimentation. Both the steady state and transient characteristics of each control algorithm along with its measured efficiency are analyzed. Finally, a novel two-mode maximum power point tracking (MPPT) control algorithm combining the constant voltage control and the incremental conduction (IncCond) methods is proposed to improve the efficiency of the 3KW PV power generation system at different insolation conditions. Experimental results show that the proposed two-mode MPPT control provides excellent performance at less than 30% insolation intensity, covering the whole insolation area without additional hardware circuitry.

Menthol biosynthesis pathway in Mentha piperita suspension cells (박하(Mentha piperita) 세포 현탁배양에서 멘톨생합성 경로)

  • Park, Si-Hyung;Chae, Young-Am;Lee, Hyong-Joo;Kim, Soo-Un
    • Applied Biological Chemistry
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    • v.36 no.5
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    • pp.358-363
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    • 1993
  • The metabolic capability of the cultured cells of peppermint was tested with whole intact cells by feeding appropriate exogenous substrates to the suspension cultures. Conversion of (-)-limonene into any other monoterpenes was not observed with the suspension cultures. (+)-Pulegone was converted into (+)-isomenthone and (-)-menthone, and (-)-menthone into (-)-menthol. The experiments confirmed that the suspension retained most of the menthol biosynthesis pathway in the cell except for a few loci. (-)-Isopiperitenone was transformed into (+)-pulegone, piperitenone, (-)-7-hydroxyisopiperitenone and two unidentified products.

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Proteome Analysis for 3T3-L1 Adipocyte Differentiation

  • Rahman, Atiar;Kumar, Suresh G.;Lee, Sung-Hak;Hyun, Sun-Hwang;Kim, Hyun-Ah;Yun, Jong-Won
    • Journal of Microbiology and Biotechnology
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    • v.18 no.12
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    • pp.1895-1902
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    • 2008
  • Adipose tissue is an important endocrine organ involved in the control of whole body energy homeostasis and insulin sensitivity. Considering the increased incidence of obesity and obesity-related disorders, including diabetes, it is important to understand thoroughly the process of adipocyte differentiation and its control. Therefore, we performed a differential proteome mapping strategy using two-dimensional gel electrophoresis combined with peptide mass fingerprinting to identify intracellular proteins that are differentially expressed during adipose conversion of 3T3-L1 pre-adipocytes in response to an adipogenic cocktail. In the current study, we identified 46 differentially expressed proteins, 6 of which have not been addressed previously in 3T3-L1 cell differentiation. Notably, we found that phosphoribosyl pyrophosphate synthetase (PRPS), a regulator of cell proliferation, was preferentially expressed in pre-adipocytes than in fully differentiated adipocytes. In conclusion, our results provide valuable information for further understanding of the adipogenic process.