• Title/Summary/Keyword: Concentration Gradient

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Two-Step Fed-Batch Culture of Recombinant Escherichia coli for Production of Bacillus licheniformis Maltogenic Amylase

  • Kim, Myoung-Dong;Lee, Woo-Jong;Park, Kwan-Hwa;Rhee, Ki-Hyeong;Seo, Jin-Ho
    • Journal of Microbiology and Biotechnology
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    • v.12 no.2
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    • pp.273-278
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    • 2002
  • Two-step fed-batch fermentations were carried out to overproduce Bacillus licheniformis maltogenic amylase (BLMA) in recombinant Escherichia coli. The first step was to increase the cell mass by controlling the feeding of a glucose solution, while the second step was designed to improve the amylase expression efficiency by supplementing organic nitrogen sources. The linear gradient feeding method was successfully adopted to maintain the glucose concentration below 0.2 g/l during the fed-batch mode, as effectively minimizing acetic acid formation. When the dissolved oxygen (DO) level became limiting, an accumulation of acetic acid and drastic decrease in specific BLMA productivity were observed. Glucose and organic nitrogen sources consisting of yeast extract and casein hydrolysate were simultaneously supplied in the pH-stat mode to further increase the specific BLMA expression efficiency. An organic nitrogen source consisting of 200 g/1 yeast extract and 100 g/1 casein hydrolysate was found to be the best among the various combinations tested. The feeding of an organic nitrogen source in the second-step fed-batch period was highly beneficial in enhancing the BLMA production. The optimized two-step fed-batch culture resulted in 78 g/l maximum dry cell mass and 443 U/ml maximum BLMA activity, corresponding to 1.5-fold increase in the dry cell mass and 3.7-fold enhancement in BLMA production, compared with the simple fed-batch fermentation.

Application of Biological Activated Carbon Process for Water Quality Improvement of Stagnant Stream Channels

  • Lee, Jae-Ho;Park, Jeung-Jin;Park, Tae-Joo;Byun, Im-Gyu
    • Environmental Engineering Research
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    • v.19 no.4
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    • pp.309-316
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    • 2014
  • The water quality improvement of golf course ponds, as representative stagnant stream channels, was evaluated by applying a biological activated carbon (BAC) process composed of four consecutive activated carbon reactors. The study was performed from autumn to winter in order to evaluate the feasibility of the BAC process under low temperature conditions. In the study, water quality of pond A (target pond) and pond B (reference pond) were monitored. Pond water was pumped into the BAC process, and was then returned to the pond after treatment. The optimal conditions were determined to be 2 hr of empty bed contact time (EBCT) at a temperature above $4^{\circ}C$, in which improvements of chemical oxygen demand (COD), total nitrogen (TN) and total phosphorus (TP) of pond A compared to pond B were 3.62%, 3.48% and 1.81%, respectively. On the other hand, as the temperature was below $4^{\circ}C$, some degree of water quality improvement was achieved even when EBCT were 1 or 0.5 hr, suggesting that the BAC process can be successfully applied for the improvement of pond water quality in winter months. The values of biomass concentration and microorganism activity in each condition were highest where 2 hr of EBCT was applied at a temperature above $4^{\circ}C$, but values were similar throughout all treatment conditions, and thus, adsorption is considered to be the dominant factor affecting process efficiency. From the denaturing gel gradient electrophoresis (DGGE) results, no significant differences were observed among the activated carbon reactors, suggesting that the number of reactors in the system could be decreased for a more compact application of the system.

ENHANCED BIOREMEDIATION AND MODIFIED BACTERIAL COMMUNITY STRUCTURE BY BARNYARD GRASS IN DIESEL-CONTAMINATED SOIL

  • Kim, Jai-Soo;Min, Kyung-Ah;Cho, Kyung-Suk;Lee, In-Sook
    • Environmental Engineering Research
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    • v.12 no.2
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    • pp.37-45
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    • 2007
  • Phytoremediation has been used effectively for the biodegradation of oil-based contaminants, including diesel, by the stimulation of soil microbes near plant roots (rhizosphere). However, the technique has rarely been assessed for itsinfluence on soil microbial properties such as population, community structure, and diversity. In this study, the removal efficiency and characteristics of rhizobacteria for phytoremediation of diesel-contaminated soils were assessed using barnyard grass (Echinochloa crusgalli). The concentration of spiked diesel for treatments was around $6000\;mg\;kg^{-1}$. Diesel removal efficiencies reached 100% in rhizosphere soils, 76% in planted bulk soils, and 62% in unplanted bulk soils after 3weeks stabilization and 2 months growth(control, no microbial activity: 32%). The highest populations of culturable soil bacteria ($5.89{\times}10^8$ per g soil) and culturable hydrocarbon-degraders($5.65{\times}10^6$ per g soil) were found in diesel-contaminated rhizosphere soil, also yielding the highest microbial dehydrogenase. This suggests that the populations of soil bacteria, including hydrocarbon-degraders, were significantly increased by a synergistic rhizosphere + diesel effect. The diesel treatment alone resulted in negative population growth. In addition, we investigated the bacterial community structures of each soil sample based on DGGE (Denaturing Gel Gradient Electrophoresis) band patterns. Bacterial community structure was most influenced by the presence of diesel contamination (76.92% dissimilarity to the control) and by a diesel + rhizosphere treatment (65.62% dissimilarity), and least influenced by the rhizosphere treatment alone (48.15% dissimilarity). Based on the number of distinct DGGE bands, the bacterial diversity decreased with diesel treatment, but kept constant in the rhizosphere treatment. The rhizosphere thus positively influenced bacterial population density in diesel-contaminated soil, resulting in high removal efficiency of diesel.

The application of DGTs for assessing the effectiveness of in situ management of Hg and heavy metal contaminated sediment

  • Bailon, Mark Xavier;Park, Minoh;Choi, Young-Gyun;Reible, Danny;Hong, Yongseok
    • Membrane and Water Treatment
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    • v.11 no.1
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    • pp.11-23
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    • 2020
  • The effectiveness of in situ sediment capping as a technique for heavy metal risk mitigation in Hyeongsan River estuary, South Korea was studied. Sites in the estuary were found previously to show moderate to high levels of contamination of mercury, methylmercury and other heavy metals. A 400 m × 50 m section of the river was selected for a thin layer capping demonstration, where the total area was divided into 4 sections capped with different combinations of capping materials (zeolite, AC/zeolite, AC/sand, zeolite/sand). Pore water concentrations in the different sites were studied using diffusive gradient in thin film (DGT) probes. All capping amendments showed reduction in the pore water concentration of the different heavy metals with top 5 cm showing %reduction greater than 90% for some heavy metals. The relative maxima for the different metals were found to be translated to lower depths with addition of the caps. For two-layered cap with AC, order of placement should be considered since AC can easily be displaced due to its relatively low density. Investigation of methylmercury (MeHg) in the site showed that MeHg and %MeHg in pore water corresponds well with maxima for sulfide, Fe and Mn suggesting mercury methylation as probably coupled with sulfate, Fe and Mn reduction in sediments. Our results showed that thin-layer capping of active sorbents AC and zeolite, in combination with passive sand caps, are potential remediation strategy for sediments contaminated with heavy metals.

Sizes and Structures of Micelles of Cationic Octadecyl Trimethyl Ammonium Chloride and Anionic Ammonium Dodecyl Sulfate Surfactants in Aqueous Solutions

  • Kim, Hong-Un;Lim, Kyung-Hee
    • Bulletin of the Korean Chemical Society
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    • v.25 no.3
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    • pp.382-388
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    • 2004
  • The sizes and structures of micelles formed in aqueous solutions of cationic octadecyl trimethyl ammonium chloride (OTAC) and anionic ammonium dodecyl sulfate (ADS) surfactants were investigated using smallangle neutron scattering (SANS), self-diffusion coefficients by pulsed-gradient spin-echo (PGSE) NMR, and dynamic light scattering (DLS) methods. SANS and DLS data indicate that their structures are spherical at concentrations as high as 300 mM. As the total surfactant concentration increases, the peaks of SANS spectra shift to higher scattering vector and become sharper, indicating that the intermicellar distance decreases and its distribution becomes narrower. This is due to more compact packing of surfactant molecules at high concentrations. The intermicellar distance of around 100 ${\AA}$ above 200 mM corresponds approximately to the diameter of one micelle. The sizes of spherical micelles are 61 ${\AA}$ and 41 ${\AA}$ for 9 mM OTAC and 10 mM ADS, respectively. Also the self-diffusion coefficients by PGSE-NMR yield the apparent sizes 96 ${\AA}$ and 31 ${\AA}$ for micelles of 1 mM OTAC and 10 mM ADS, respectively. For ADS solutions of high concentrations (100-300 mM), DLS data show that the micelle size remains constant at $25{\pm}2{\AA}$. This indicates that the transition in micellar shape does not take place up to 300 mM, which is consistent with the SANS results.

Mitigating $CH_4$ Emissions in Semi-Aerobic Landfills: Impacts of Operating Conditions on Abundance and Community Structure of Methanotrophs in Cover Soils

  • Li, Huai;Chi, Zi-Fang;Lu, Wen-Jing;Wang, Hong-Tao
    • Journal of Microbiology and Biotechnology
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    • v.23 no.7
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    • pp.993-1003
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    • 2013
  • Methanotrophs are the most important sink of $CH_4$, which is a more highly potent greenhouse gas than $CO_2$. Methanotrophic abundance and community diversity in cover soils from two typical semi-aerobic landfills (SALs) in China were detected using real-time polymerase chain reaction (real-time-PCR) and denaturing gradient gel electrophoresis (DGGE) based on 16S rRNA genes, respectively. Real time-PCR showed that Type I methanotrophs ranged from $1.07{\times}10^6$ to $2.34{\times}10^7$ copies/g soil and that of Type II methanotrophs from $1.51{\times}10^7$ to $1.83{\times}10^8$ copies/g soil. The ratio of Type II to Type I methanotrophic copy numbers ranged from 5.61 to 21.89, indicating that Type II methanotrophs dominated in SAL. DGGE revealed that Type I methanotrophs responded more sensitively to the environment, changing as the community structure varied with different soil types and locations. Methylobacter, Methylosarcina, and Methylomicrobium for Type I, and Methylocystis for Type II were most prevalent in the SAL cover layer. Abundant interflow $O_2$ with high $CH_4$ concentration in SALs is the reason for the higher population density of methanotrophs and the higher enrichment of Type II methanotrophs compared with anaerobic landfills and other ecosystems, which proved a conclusion that increasing the oxygen supply in a landfill cover layer would greatly improve $CH_4$ mitigation.

Retention Behavior of Lanthanide Complexes with $\alpha$ -hydroxyisobutyric Acid on Cation Exchanger (양이온 교환체에서 희토류원소와 $\alpha$-Hydroxyisobutyric Acid 착물들의 머무름 거동에 관한 연구)

  • Jo, Gi Su;Han, Seon Ho;Seo, Mu Yeol;Eom, Tae Yun;Kim, Yeon Du
    • Journal of the Korean Chemical Society
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    • v.34 no.6
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    • pp.582-592
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    • 1990
  • Retention behavior of lanthanide-$\alpha$HiBA complexes was studied on the cation exchanger (LC-18 coated with $C_{20}H_{41}SO_4^-$). An equation predicting retention of lanthanides in isocratic or gradient elution with sodium ion and $\alpha$-HiBA concentration was derived from ion exchange equilibria of metal-ligand complex system, respectively. The relations between log k' and log [Na$^+$] /log [$\alpha$-HiBA) showed non-linearity in isocratic elution. In gradient elution a good linearity between log k' vs log R was obtained. The values of slopes (log k / log R) gave good agreements between calculation and experiment. Individual capacity factors ($k'_{Ln}^{3+}, k'_{LnL}^{2+}, k'{LnL2+}) and stability constant (${\beta}_1$, ${\beta}_2$, ${\beta}_3$) of lanthanide-$\alpha$HiBA complexes were calculated by the non-linear least square fittings using the retention equation. The correlation coefficients of lanthanides were shown better than 0.9996 between experiment and calculation.

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Simultaneous Determination of Catecholamines, Serotonin and Their Metabolites in the Biological Sample Using HPLC/ECD (생체 시료 중 카테콜 아민류, 세로토닌 및 대사물질들의 HPLC/ECD 동시 정량분석)

  • Min, Ji-Hyun;Hahn, Young-Hee
    • YAKHAK HOEJI
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    • v.55 no.4
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    • pp.277-283
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    • 2011
  • Simultaneous monitoring of catecholamines and serotonin with their appropriate extraction from the biological samples is required in order to understand thoroughly the regulation of the central and peripheral nervous system. In the present research the segmented gradient elution with the solid phase extraction using a C18 cartridge rather than the previous isocratic elution with alumina extraction is successfully employed to determine norepinephrine (NE), epinephrine (E), dopamine (DA), serotonin (5HT), 3,4-dihydroxyphenylacetic acid (DOPAC) and 5-hydroxyindoleacetic acid (5HIAA) simultaneously within 20 minutes using 3,4-dihydroxyhydrocinnamic aicd as the internal standard (IS). Linearities were obtained in the concentration range between $5{\times}10^{-6}M$ and $1{\times}10^{-4}M$ for all 7 compounds with detection limits of 0.6~1.9 ${\mu}M$. The present HPLC/ECD method yielded reasonable accuracy (relative error; -1.4~1.1%) and precision (relative standard deviation; 0.4~1.9%) for 9 measurements of the standard solution consisting of NE, E, DA, 5HT, DOPAC and 5HIAA compounds. Recoveries of catecholamines, serotonin and their metabolites from human serum were in the range of 57%~86%. While the concentrations of NE and 5HT in the serum of normal Sprague-Dawley rat were found as $1.4{\times}10^{-6}M$ and $2.6{\times}10^{-6}M$, respectively, the contents of NE and 5HT in the serum of the stressed rat were increased 5.6 times and 1.4 times more, respectively.

Ultra-fast Generic LC-MS/MS Method for High-Throughput Quantification in Drug Discovery

  • Kim, So-Hee;Yoo, Hye Hyun;Cha, Eun-Ju;Jeong, Eun Sook;Kim, Ho Jun;Kim, Dong Hyun;Lee, Jaeick
    • Mass Spectrometry Letters
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    • v.4 no.3
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    • pp.47-50
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    • 2013
  • An ultra-fast generic LC-MS/MS method was developed for high-throughput quantification of discovery pharmacokinetic (PK) samples and its reliability was verified. The method involves a simple protein precipitation for sample preparation and the analysis by ultra-fast generic LC-MS/MS with the ballistic gradient program and selected reaction monitoring (SRM) mode. Approximately 290 new chemical entities (NCEs) (over 10,000 samples) from 5 therapeutic programs were analyzed. The calibration curves showed good linearity in the concentration range of 1, 2 or 5 to 2000 ng/mL. No significant ion suppression was observed in the elution region of all the NCEs. When approximately 300 plasma samples were continuously analyzed, the peak area of internal standard was constant and reproducible. In the repeated analysis of samples, the plasma concentrations and the area under the curve (AUC) were consistent with the results from the first analysis. These results showed that the present ultra-fast generic LC-MS/MS method is reliable in terms of selectivity, sensitivity, and reproducibility and could be useful for high-throughput quantification and other bioanalysis in drug discovery.

Mechanical and thermodynamic stability, structural, electronics and magnetic properties of new ternary thorium-phosphide silicides ThSixP1-x: First-principles investigation and prospects for clean nuclear energy applications

  • Siddique, Muhammad;Iqbal, Azmat;Rahman, Amin Ur;Azam, Sikander;Zada, Zeshan;Talat, Nazia
    • Nuclear Engineering and Technology
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    • v.53 no.2
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    • pp.592-602
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    • 2021
  • Thorium compounds have attracted immense scientific and technological attention with regard to both fundamental and practical implications, owing to unique chemical and physical properties like high melting point, high density and thermal conductivity. Hereby, we investigate the mechanical and thermodynamic stability and report on the structural, electronic and magnetic properties of new silicon-doped cubic ternary thorium phosphides ThSixP1-x (x = 0, 0.25, 0.5, 0.75 and 1). The first-principles density functional theory procedure was adopted within full-potential linearized augmented plane wave (FP-LAPW) method. The exchange and correlation potential terms were treated within Generalized-Gradient-Approximation functional modified by Perdew-Burke-Ernzerrhof parameterizations. The proposed compounds showed mechanical and thermodynamic stable structure and hence can be synthesized experimentally. The calculated lattice parameters, bulk modulus, total energy, density of states, electronic band structure and spin magnetic moments of the compounds revealed considerable correlation to the Si substitution for P and the relative Si/P doping concentration. The electronic and magnetic properties of the doped compounds rendered them non-magnetic but metallic in nature. The main orbital contribution to the Fermi level arises from the hybridization of Th(6d+5f) and (Si+P)3p states. Reported results may have potential implications with regard to both fundamental point of view and technological prospects such as fuel materials for clean nuclear energy.