• Title/Summary/Keyword: Immobilized cell

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Rapid Detection of Salmonella spp. by Antibody-Immobilized Piezoelectric Crystal Biosensor (고정화법을 달리하여 제조한 압전류적 항체 센서에 의한 Salmonella spp.의 신속 검출)

  • 박인선;김우연;김남수
    • Journal of Food Hygiene and Safety
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    • v.13 no.3
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    • pp.206-212
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    • 1998
  • An improved antibody-coated sensor system based on quartz crystal microbalance was developed for the detection of Salmonella spp. An antibody against Salmonella common structural antigen was immobilized onto one gold electrode of the piezoelectric quartz crystal surface by various immobilization procedures. The best results in sensitivity and stability were obtained with the thin layers of protein A and 3,3'-dithiopropionimidate.2HCI(DTBP), a homobifunctional thiol-cleavable crosslinker. After the addition of a S. typhimurium suspension into a reaction cell with 0.1 M sodium phosphate buffer, pH 7.2, the resonant frequency owing to S. typhimurium adsorption decreased conspicuously. The antibody-immobilized crystals prepared by the gold-protein A complex formation and DTBP thiolation showed the frequency shifts of 80 and 283 Hz, respectively. The time required for maximum frequency shift was about 30~60 min. The antibody-coated crystal could be reused for 6~8 consecutive assays.

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Long-term Repeated-Batch Operation of Immobilized Escherichia coli Cells to Synthesize Galactooligosaccharide

  • Lee, Sang-Eun;Yeon, Ji-Hyeon;Jung, Kyung-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.22 no.11
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    • pp.1486-1493
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    • 2012
  • In this study, we investigated whether galactooligosaccharide (GOS) can be stably and steadily synthesized using immobilized ${\beta}$-galactosidase (${\beta}$-gal) inclusion body (IB)-containing E. coli cells during long-term repeated-batch operation. To improve the operational stability of this enzyme reactor system, immobilized E. coli cells were crosslinked with glutaraldehyde (GA) after immobilization of the E. coli. When we treated with 2% GA for E. coli crosslinking, GOS production continued to an elapsed time of 576 h, in which seven batch runs were operated consecutively. GOS production ranged from 51.6 to 78.5 g/l ($71.2{\pm}10.5$ g/l, n = 7) during those batch operations. In contrast, when we crosslinked E. coli with 4% GA, GOS production ranged from 31.5 to 64.0 g/l ($52.3{\pm}10.8$, n = 4), and only four consecutive batch runs were operated. Although we did not use an industrial ${\beta}$-gal for GOS production, in which a thermophile is used routinely, this represents the longest operation time for GOS production using E. coli ${\beta}$-gal. Improved stability and durability of the cell immobilization system were achieved using the crosslinking protocol. This strategy could be directly applied to other microbial enzyme reactor systems using cell immobilization to extend the operation time and/or improve the reactor system stability.

Solvent effect on enzymatic steroid transformation

  • Kim, Doo-Ha;Lee, S.B.;Ryu, D.Y.
    • Archives of Pharmacal Research
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    • v.3 no.1
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    • pp.1-6
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    • 1980
  • As part of our endeavor to increase the productivity of steroid by enzymatic transformation of corticosteroids, attempts have been made to increase the solubility of steroids by using some organic solvents. When the solubility of steroids is the rate limiting factor in the steroid transformation, it was found that the use of solvents significantly improved the yield. Hydrocorisone as a substrate and 3-ketosteroid .DELTA.$^{1}$ dehydrogense as an immobilized whole cell enzyme were employed as the model system for this study. It was found that the yield of product, prodnisolone, goes through a maximum with an increase in the solvent concentration. At a high solvent concentration, the solvent showed a toxic effect and it causes a decrease in the product yield by the second order inhibition mechanism. Among the solvents evaluated, methanol and ethanol were found to be the best. These alcohols are not only good solvents but also showed minimal toxic effect. Based on the experimental results, it was concluded that the productivity of steroid can be increased by usign well selcted solvents systems for the enzymatic transformation of steroids.

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High Density Cell Cultivation of Escherichia coli in a Dual Hollow Fiber Bioreactor (이중실관 반응기에서 E. coli의 고농도 배양)

  • Chung, Bong-Hyun;Chang, Ho-Nam;Kim, In-Ho
    • Microbiology and Biotechnology Letters
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    • v.13 no.3
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    • pp.209-212
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    • 1985
  • The cell density and packing characteristics of Escherichia coli immobilized in a dual hollow fiber bioreactor consisting of outer silicone membrane for oxygen transport and three inner isotropic polypropylene hollow fibers for substrate transport were investigated. The cells have grown forming the layer like animal tissue in a nearly 100% packing density. The dry biomass density was 550g/liter of void volume for cell growth, which was the highest among the biomass densities ever reported.

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Algicidal Effect of Immobilized Bacteria against S. hantzschii in Microcosm (살조세균 Pseudomonas fluorescens HYK0210-SK09의 두 가지 담체 포집능과 이를 이용한 microcosm에서 Stephanodiscus hantzschii (Bacillariophyceae)의 살조능 연구)

  • Jung, Seung-Won;Kim, Young-Ok;Han, Myung-Soo
    • Korean Journal of Environmental Biology
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    • v.27 no.3
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    • pp.244-251
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    • 2009
  • To assess the algicidal effect of a bacterium, Pseudomonas fluorescens HYK0210-SK09 (SK09), attached to activated carbon polyvinyl alcohol (ACPA) and cellulose sponge (CS) carriers against Stephanodiscus hantzschii, the present study was carried out in an indoor-microcosm. As comparing immobilization effects of two carriers, the ACPA carrier allowed for higher packing cell density of SK09 compared to the CS carrier. In the microcosm, immobilized SK09 cells were applied to control S. hantzschii blooms. Immobilized SK09 cells exhibited a species-specific activity towards the diatom, showing an algicidal effect up to 72% attached by ACPA carriers and to 51% attached by CS carriers. In particular, a level of conductivity treated with ACPA carriers was decreased than that of CS carriers. The present study clearly demonstrates that ACPA-immobilized SK09 cells could effectively control S. hantzschii blooms and improve water quality in the microcosm ecosystem.

Performance Comparison of Continuous Reactors for Bioethanol Production Based on Glycerol (글리세롤 기반의 바이오에탄올 생산을 위한 연속생산반응기의 성능 비교)

  • Lee, Sang-Jun;Song, Yoon-Seok;Kim, Sung-Bong;Kang, Sung-Woo;Han, Sung-Ok;Park, Chul-Hwan;Kim, Seung-Wook
    • KSBB Journal
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    • v.26 no.4
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    • pp.328-332
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    • 2011
  • Ethanol production using glycerol as a carbon source was performed by Enterobacter aerogenes immobilized on calcium alginate beads. To improve the ethanol production, the optimal conditions such as loading amount of immobilized cells and glycerol concentration were investigated. The optimal loading amount of immobilized cells and glycerol concentration were 10 mL of calcium alginate bead and 10 g/L, respectively. Consequently, glycerol consumption rate, ethanol concentration and yield were 0.32 g/$L{\cdot}h$, 3.38 g/L and 0.43 g/g on the batch production, respectively. Continuous production of ethanol was successfully achieved using two types of immobilized cell reactors (continuous stirred tank reactor and packed bed reactor) from 10 g/L of glycerol. In the continuous stirred tank reactor, glycerol consumption, ethanol concentration, specific productivity and yield were 9.8 g, 4.67 g/L, 1.17 g/$L{\cdot}h$, 0.48 g/g, respectively. The concentration of produced ethanol was 38-44% higher comparison to batch fermentation, and continuous stirred tank reactor showed better performance than packed bed reactor.

Photocatalytic Cell Disruption of Giardia lamblia in a $UV/TiO_2$ Immobilized Optical-Fiber Reactor

  • YU , MI-JIN;KIM, BYUNG-WOO
    • Journal of Microbiology and Biotechnology
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    • v.14 no.6
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    • pp.1105-1113
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    • 2004
  • Disinfection of a waterborne pathogenic protozoa, Giardia lamblia, by the conventional chlorine method has been known to be difficult. An alternative disinfection has been carried out by using a UV -light illuminating optical­fiber photoreactor. Light intensity diffused from one piece of a clad-removed optical-fiber was $1- 1.5{\mu}Em^{-2}s^{-1}$. Disinfection capability in a UV -light irradiated optical-fiber reactor suspended with 0.01 g $TiO_{2}\;dm^{-3}$ was 1.4 times that in the same reactor without $TiO_{2}$ photocatalysts. To resolve the absorption and scattering of UV light by the particles themselves as well as the difficulty of recycling particles in the slurry­type reactor, $TiO_{2}$ which was obtained by a hydrothermal method, was immobilized on clad-removed optical fibers. Such pretreatment of fiber surface resulted in an excellent transparency, which enhanced the UV light to diffuse laterally from a fiber surface. Coating time of the prepared solution by the hydrothermal method was not effective after more than two times. Disinfection capability in the $TiO_{2}$-immobilized optical-fiber reactor was $83\%$ in 1 h at $40^{\circ}C$, which was slightly higher than $76\%$ at $22^{\circ}C$ and $68\%$ at $10^{\circ}C$. Disinfection capability at $22^{\circ}C$ increased from $74\%$ at an initial pH of 3.4, through $76\%$ at pH 6.5, to $87\%$ at an initial pH of 10. Oxygen supply with air-flow rate of 5 $cm^3\;min^{-1}$ did not seem to increase the disinfection capability with UV /immobilized $TiO_2$.

Production of ${\gamma}$-Glutamylcysteine by Immobilized Mixed Microbial System of Recombinant E. coli and Yeast (재조합 대장균과 효모의 고정화 혼합세포계에 의한 ${\gamma}$-Glutamylcysteine 생산)

  • 김원근;구윤모
    • KSBB Journal
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    • v.10 no.3
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    • pp.249-256
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    • 1995
  • ${\gamma}$-Glutamylcysteine production by the immobilized microbial system of recombinant Escherichia coli and yeast was investigated. ${\gamma}$-Glutamylcysteine was synthesized from L-glutamic acid and L-cysteine in the presence of ATP by the reaction catalyzed by ${\gamma}$-glutamylcysteine synthetase. An immobilized microbial cell system was developed for the efficient ${\gamma}$-glutamylcysteine production. Recombinant Escherichia coli and yeast were immobilized by alginate. Production of ${\gamma}$-glutamylcysteine was better with the recombinant Escherichia coli for both the synthesis of ${\gamma}$-glutamylcysteine and the ATP regeneration than the mixed system of recombinant Escherichia coli and yeast. The proper radio of recombinant Escherichia coli to yeast was experimentary observed to be 1:4 in the mixed system. Although the immobi1ized system had the slower reaction rate, its reaction stability was increased by 10%.

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Isolation, identification and immobilized-cell characteristics of a bacterium that produces $N_2$ from $NH{_4}{^+}$ under an aerobic condition

  • Park, Kyoung-Joo;Cho, Kyoung-Sook;Kim, Jeong-Bo;Lee, Min-Gyu;Lee, Byung-Hun;Hong, Young-Ki;Kim, Joong-Kyun
    • 한국생물공학회:학술대회논문집
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    • 2005.10a
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    • pp.450-455
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    • 2005
  • To treat wastewater efficiently by a one-step process of nitrogen removal, a new strain of $N_2-producing$ bacteria from $NH{_4}{^+}$ under an aerobic condition was isolated and identified. By 16S-rDNA analysis, the isolate was identified as Enterobacter asburiae with 96% similarity. The isolate shows that the capacity of $N_2$ production under an oxic condition was approximately three times higher than that under an anoxic condition. The optimal conditions (pH, temperature and C/N ratio) of the immobilized isolate for $N_2$ production were found to be 7.0, $30^{\circ}C$ and 5, respectively. Under all the optimum reaction conditions, the removal efficiency of $COD_{Cr}$ and TN reached 56.1 and 60.9%, respectively. The removal rates of $COD_{Cr}$ and TN were highest for the first 2.5 hrs (with the removal $COD_{Cr}$ ratios of 32.1), and afterwards the rates decreased as reaction proceeded. For application of the immobilized isolate to a practical process of ammonium removal, a continuous bioreactor system exhibited a satisfactory performance at HRT of 12.1 hr, in which the effluent concentrations of $NH{_4}{^+}-N$ was measured to be 15.4 mg/L with its removal efficiency of 56.0%. The maximum removal rate of $NH{_4}{^+}-N$ reached 1.6 mg $NH{_4}{^+}-N/L/hr$ at HRT of 12.1 hr (with N loading rate of 0.08 $Kg-N/m^3-carrier/d)$. As a result, the application of the immobilized isolate appears a viable alternative to the nitrification-denitrification processes.

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