• Title/Summary/Keyword: NH_3$

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Biochemical Characterization of Recombinant L-Asparaginase (AnsA) from Rhizobium etli, a Member of an Increasing Rhizobial-Type Family of L-Asparaginases

  • Moreno-Enriquez, Angelica;Evangelista-Martinez, Zahaed;Gonzalez-Mondragon, Edith G.;Calderon-Flores, Arturo;Arreguin, Roberto;Perez-Rueda, Ernesto;Huerta-Saquero, Alejandro
    • Journal of Microbiology and Biotechnology
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    • v.22 no.3
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    • pp.292-300
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    • 2012
  • We report the expression, purification, and characterization of L-asparaginase (AnsA) from Rhizobium etli. The enzyme was purified to homogeneity in a single-step procedure involving affinity chromatography, and the kinetic parameters $K_m$, $V_{max}$, and $k_{cat}$ for L-asparagine were determined. The enzymatic activity in the presence of a number of substrates and metal ions was investigated. The molecular mass of the enzyme was 47 kDa by SDS-PAGE. The enzyme showed a maximal activity at $50^{\circ}C$, but the optimal temperature of activity was $37^{\circ}C$. It also showed maximal and optimal activities at pH 9.0. The values of $K_m$, $V_{max}$, $k_{cat}$, and $k_{cat}/K_m$ were $8.9{\pm}0.967{\times}10^{-3}$ M, $128{\pm}2.8$ U/mg protein, $106{\pm}2s^{-1}$, and $1.2{\pm}0.105{\times}10^4M^{-1}s^{-1}$, respectively. The L-asparaginase activity was reduced in the presence of $Mn^{2+}$, $Zn^{2+}$, $Ca^{2+}$, and $Mg^{2+}$ metal ions for about 52% to 31%. In addition, we found that $NH_4{^+}$, L-Asp, D-Asn, and ${\beta}$-aspartyl-hydroxamate in the reaction buffer reduced the activity of the enzyme, whereas L-Gln did not modify its enzymatic activity. This is the first report on the expression and characterization of the L-asparaginase (AnsA) from R. etli. Phylogenetic analysis of asparaginases reveals an increasing group of known sequences of the Rhizobial-type asparaginase II family.

Effect of Saline Concentrations on Biological Nitrification in Batch Reactor

  • Lee, Young Joon;Nguyen, Viet Hoang;Nguyen, Hong Khanh;Pham, Tuan Linh;Kim, Gi Youn
    • Journal of Integrative Natural Science
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    • v.4 no.2
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    • pp.103-112
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    • 2011
  • This study was carried out on 4 batch reactors to determine the specific ammonium oxidizing rate (SAOR), specific nitrate forming rate (SNFR) and inhibitory degree of nitrifying activities with saline concentrations. Under salt free condition ammonia was consumed during the reaction period within 200 min. When the salt level increased to 10, 20 and 30 g $NaClL^{-1}$ in reactor, ammonia depletion took 250, 300 and above 350 min, respectively. During concentration above 10 g $NaClL^{-1}$, there was nitrite accumulation. Also, at 30 g $NaClL^{-1}$ ammonia did not depleted and $NO_2{^-}$-N accumulated until the final reaction. Nitrate formation rates decreased with increasing salt concentration. SAOR and SNFR showed a decreasing trend as salinity concentrations were increased. The SAOR was reduced from 0.2 to 0.08 mg $NH_4{^+}$-N $g^{-1}VSS\;day^{-1}$ as the salt concentration increased from 0 to 30 g $NaClL^{-1}$. Similarly, the SNFR decreased from 0.26 kg $NO_3{^-}$-N $kg^{-1}VSS\;day^{-1}$ at saline free to 0.1 kg $NO_3{^-}$-N $kg^{-1}VSS\;day^{-1}$ at saline 30 g L-1. A severe inhibition of nitrifiers activity was observed at increased salt concentrations. The inhibition ratio of specific ammonium oxidation rates were 17, 47 and 60% on the reactor of 10, 20 and 30 g $NaClL^{-1}$ added, respectively. The inhibition ratio of specific nitrate forming rates also were inhibited 30, 53 and 62% on the reactor of 10, 20 and 30 g $NaClL^{-1}$ added, respectively. As the salinity concentrations increased from 0 to 30 mg $NaClL^{-1}$, the average MLSS concentration increased from 1,245 to 1,735 $mgL^{-1}$. The SS concentration of supernatant in reactor which settled about 30 minutes was not severely difference between concentration of salt free reactor and one of those high salt contained reactors.

Preparation of LiFe PO4 Using Chitosan and its Cathodic Properties for Rechargeable Li-ion Batteries

  • Hong, Kyong-Soo;Yu, Seong-Mi;Ha, Myoung-Gyu;Ahn, Chang-Won;Hong, Tae-Eun;Jin, Jong-Sung;Kim, Hyun-Gyu;Jeong, Euh-Duck;Kim, Yang-Soo;Kim, Hae-Jin;Doh, Chil-Hoon;Yang, Ho-Soon;Jung, Hee
    • Bulletin of the Korean Chemical Society
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    • v.30 no.8
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    • pp.1719-1723
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    • 2009
  • The LiFeP$O_4$ powder was synthesized by using the solid state reaction method with Fe($C_2O_4){\cdot}2H_2O,\;(NH_4)_2HPO_4,\;Li_2CO_3$, and chitosan as a carbon precursor material for a cathode of a lithium-ion battery. The chitosan added LiFePO4 powder was calcined at 350 ${^{\circ}C}$ for 5 hours and then 800 ${^{\circ}C}$ for 12 hours for the calcination. Then we calcined again at 800 ${^{\circ}C}$ for 12 hours. We characterized the synthesized compounds via the crystallinity, the valence states of iron ions, and their shapes using TGA, XRD, SEM, TEM, and XPS. We found that the synthesized powders were carbon-coated using TEM images and the iron ion is substituted from 3+ to 2+ through XPS measurements. We observed voltage characteristics and initial charge-discharge characteristics according to the C rate in LiFeP$O_4$ batteries. The obtained initial specific capacity of the chitosan added LiFeP$O_4$ powder is 110 mAh/g, which is much larger than that of LiFeP$O_4$ only powder.

Amperometric Determination of Histamine using Immobilized Enzyme Reactors with Different Carriers (담체 고정화 효소 반응기를 이용한 Histamine의 전기화학적 측정)

  • Ji, Jung-Youn;Jeon, Yeon-Hee;Kim, Mee-Ra
    • Journal of the East Asian Society of Dietary Life
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    • v.22 no.1
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    • pp.88-94
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    • 2012
  • Histamine is a kind of primary biogenic amine arising from the decarboxylation of the amino acid L-histidine. The toxicology of histamine and its occurrence and formation in foods are especially emphasized in fermented foods. In this study, the biosensor for detection of histamine with functionalized multi-walled carbon nanotubes (MWCNT) was developed. We also searched for an appropriate insoluble substrate to immobilize the enzyme. The developed biosensor showed a detection limit of $0.1{\mu}M$ hydrogen peroxide. The enzyme reactor was prepared with diamine oxidase immobilized on insoluble carriers including CNBr-activated sepharose 4B, calcium alginate, and controlled pore size glass beads. The coupling efficiency of CNBr-activated sepharose 4B, calcium alginate, and controlled pore size glass beads were 48.5%, 40.3%, and 51.0%, respectively. In addition, the response currents on histamine with each immobilized enzyme reactor prepared with CNBr-activated sepharose 4B, calcium alginate, and controlled pore size glass beads were 120 nA, 110 nA, and 140 nA at $100{\mu}M$ of histamine concentration, respectively. Therefore, it is suggested that controlled pore size glass beads are the best carriers for immobilizing diamine oxidase to detect histamine in this biosensor.

Advanced Wastewater Treatment on ship's sewage with new MARPOL73/78 ANNEX IV (MARPOL73/78 ANNEX IV의 개정발효에 따른 선박 오수의 고도처리)

  • Park, Sang-Ho;Lim, Jae-Dong;Park, Sung-Jeng;Kim, In-Soo
    • Proceedings of KOSOMES biannual meeting
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    • 2007.05a
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    • pp.91-96
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    • 2007
  • Lab scale experimental study was carried out for SBR process, to investigate the effects of influent ship sewage organic compound removal and Bacillus sp. state on design parameters. This process was able to remove nitrogen and phosphorus as well as organic matter efficiently. More than 95% of chemical oxygen demand(COD) were removed. In addition, about 97% of total nitrogen (T-N) was reduced. The total phosphorus(T-P) reduction averaged 93%. The performance load of SBR process was shown to be $0.095kg{\cdot}TOC/m^3{\cdot}day$. The pH was decreased from 8.1 to 7.0 within 30 min and increased to 7.3 at the end of anoxic stage, and these phenomena were explained. The sludge produced in the SBR process is characterized by low generation rate (about $0.36kg{\cdot}MLSS/kg{\cdot}TOC$) and excellent settleability. The number of Bacillus sp. in the SBR was 24.2%, indicating that Bacillus sp. was a predominant species in the reactor.

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Inhibitory Effect of Extract from Ostericum koreanum on LPS-induced Proinflammatory Cytokines Production in RAW264.7 Cells (LPS로 자극한 RAW264.7 세포에서 강활 추출물의 염증성세포활성물질의 억제효과)

  • Park, Hee-Je;Bae, Gi-Sang;Kim, Do-Yun;Seo, Sang-Wan;Park, Kyung-Bae;Kim, Byung-Jin;Song, Je-Moon;Lee, Kyung-Yong;Na, Chul;Shin, Byung-Chul;Park, Sung-Joo;Song, Ho-Joon;Hwang, Sung-Yeon
    • The Korea Journal of Herbology
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    • v.23 no.3
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    • pp.127-134
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    • 2008
  • Objectives : The present study was designed to investigate whether Ostericum koreanum (OK) could regulate lipopolysaccharide (LPS)-induced inflammatory response in vitro and in vivo. Methods : To evaluate of anti-inflammatory effect of OK, we examined Nitric oxide (NO), proinflammatory cytokines production in LPS-stimulated RAW264.7 cells. Furthermore, we checked molecular mechanism especially in the phosphorylation of mitogen-activated protein kinases (MAPKs) and the degradation of inhibitory kappa B a ($Ik-B{\alpha}$) using western blot and also investigated survival of mice in LPS-mediated endotoxin shock. Results : 1. Extract from OK itself have weak cytotoxic effect on RAW264.7 cells. Extract from OK inhibited LPS-induced NO, tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), interleukin $(IL)-1{\beta}$, IL-6 and IL-10 production in RAW264.7 cells. 2. OK inhibited the phosphorylation of MAPKs, such as p38, extracelluar signal-regulated kinase (ERK1/2) and c-Jun NH2-terminal kinase (JNK) and also the degradation of $I{\kappa}-B{\alpha}$ in the LPS-stimulated RAW264.7 cells 3. OK did not inhibit LPS-induced endotoxin shock. Conclusions : OK down-regulated LPS-induced NO and cytokines production through suppressing activation of MAPKs and degradation of $I{\kappa}-B{\alpha}$. Our results suggested that OK may be a beneficial drug against inflammatory diseases.

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Suitability evaluation of magnesium substituted biphasic calcium phosphates prepared by coprecipitation method (공침법을 이용한 마그네슘이 첨가된 biphasic calcium phosphate의 적합성 평가)

  • Lee, Hyoung-Sin;Kim, Tae-Wan;Kim, Dong-Hyun;Park, Hong-Chae;Yoon, Seog-Young
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.20 no.5
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    • pp.237-242
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    • 2010
  • Magnesium-substituted BCP (biphasic calcium phosphate) powders were prepared by incorporating small amounts of magnesium into the structure of different hydroxyapatite (HAp)/${\beta}$-tricalcium phosphate (${\beta}$-TCP) ratios through coprecipitation method. A series of magnesium substitutions ranging from 0, 0.5, and 1.0 wt%, which are comparable to the measured magnesium contents, were performed. The obtained powders were characterized by the following analytical techniques: X-ray diffraction analysis (XRD), Thermo Gravimetric Analyzer (TGA) and Fourier transform infrared spectroscopy (FT-IR). The results have shown that substitution of magnesium in the calcium-deficient apatites resulted in the formation of biphasic mixtures of different HAP/${\beta}$-TCP ratios after heating above $1000^{\circ}C$. The 1.0 wt% magnesiumsubstituted-BCP were soaked in Hank's solutions after 2 weeks to observe the morphology of the biocement, especially needle-like hydroxyapatite crystals and to estimate the length and diameter of nanoneedle crystals.

Comparison of Biochemical Characterization of Korean and Chinese Mung Bean Lectin (한국산 녹두와 중국산 녹두에 있어서 Lectin의 생화학적 특성 비교)

  • Roh, Kwang Soo
    • Journal of Life Science
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    • v.24 no.6
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    • pp.603-611
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    • 2014
  • The lectins were separated from Korean and Chinese mung bean seeds finally via chromatography using Sephadex G-100 and their biochemical features were studied and compared. They showed no hemagglutination with human red blood cells regardless of trypsin treatment and showed hemagglutination with only trypsin treated rabbit red blood cells. The molecular weights of two lectins were identified as 54 kDa and 28 kDa by SDS-PAGE. It was found that while the optimal reaction temperature of the lectin from Korean mung bean was $60^{\circ}C$, that of the lectin from Chinese mung bean seeds was $50^{\circ}C$. It was found also that the most thermal stable temperature of the seed lectin from Korean mung bean seeds was $50^{\circ}C$ and the lectin from Chinese mung bean was $40-50^{\circ}C$. The lectin from Korean mung bean seeds showed the highest activity at pH 3.2 and the lectin from Chinese mung bean showed the highest activity at pH 6.2. It was identified that when treating a denaturant, thiourea and guanidine-HCl resulted in no hemagglutination, so they induced denaturalization. It was identified also that there was no hemagglutination with urea, so it did not induced denaturalization. They showed no septicity to 6 types of carbohydrates including D-glucose. In addition, the lectins from the two mung bean seed had specificity to metal ions.

A Study on the Thermal Decomposition and Injection Direction of Urea Solution Used in DeNOx Process (탈질공정에 사용되는 우레아 수용액의 열분해와 분사방향에 관한 연구)

  • Moon, Seung-Hyun
    • Journal of Korean Society of Environmental Engineers
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    • v.31 no.7
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    • pp.531-540
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    • 2009
  • In this study, thermo-gravimetric analysis(TGA) was used to investigate the effect of urea concentration and heating rate on the ammonia($NH_3$) formation process from urea solution. A newly designed pipe nozzle was inserted through a 1,000 N${\ss}$(C)/h oil firing boiler to compare the DeNOx efficiencies between the upward and downward nozzle. This experiment reveals the effect of path which an urea droplet goes through. Urea solution showed the same TGA graph without regard to the presence of oxygen. Heating rate had a great influence on the weight loss trend. But the concentration of urea solution between 10% and 40% did not affect so much the thermal decomposition temperature. Therefore, heating rate is more important factor on the thermal decomposition of urea than the concentration of urea solution. Three nozzles located at different positions showed similar DeNOx efficiencies such as 68.1%, 71.8%, 70.8% at the same temperature. Even though urea solution was injected for the same zone, the injection direction made much difference in DeNOx efficiency. A upward nozzle showed 68.1% and downward nozzle 9.5%. This results illustrate the importance of heating rate.

Development and Application of Water Quality Level Model (WQLM) for the Small Streams of Rural Watersheds with Discriminant Analysis (판별분석을 통한 농촌유역 소하천의 수질등급모형(WQLM) 개발 및 적용)

  • Kim, Jin-Ho;Choi, Chul-Mann;Ryu, Jong-Soo;Jung, Goo-Bok;Shin, Joung-Du;Han, Kuk-Heon;Lee, Jung-Taek;Kwun, Soon-Kuk
    • Journal of Korean Society on Water Environment
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    • v.23 no.2
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    • pp.260-265
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    • 2007
  • This study was carried out to complement water quality standards and to establish new concept for water quality standards reflecting current state of water quality in small streams. By this purpose, discriminant analysis was performed and Water Quality Level Model (WQLM) was developed using the data such as EC, BOD, $COD_{Mn}$, SS, T-N, T-P, $NH_3-N$ in 224 agricultural streams. To give water quality level for water quality parameters, it divided into 20% respectively in the order of excellent water quality. On the basis of the lowest water quality level, water quality level of small streams is granted. As a result of it, number of stream corresponding to Level I was no, Level II was 2 streams, Level III was 22 streams, Level IV was 70 streams, and Level V was 130 streams. Average of water quality in each level was the highest in Level V. EC, SS, and T-N of 7 parameters were selected in variance concerned water quality level. By standardized canonical discriminant function coefficient, EC of three variances was the highest in 0.625 at the discriminant power. The next was T-N (0.509), SS (0.414). By discriminant function for water quality level, Level II was equal to $-2.973+19.376{\times}(EC)+0.647{\times}(T-N)+0.009{\times}(SS)$, Level III was equal to $-3.288+19.190{\times}(EC)+0.733{\times}(T-N)+0.041{\times}(SS)$, Level IV was equal to $-4.462+27.097{\times}(EC)+0.792{\times}(T-N)+0.053{\times}(SS)$, and Level V was equal to $-9.117+40.040{\times}(EC)+1.305{\times}(T-N)+0.111{\times}(SS)$. As a result of test at real agricultural watershed of Jeongan and Euidang in Gongju city, the fitness of WQLM was high to 88.78%. But, to get accomplished water quality assessment more exactly in agricultural streams, we had to concentrate and get vast data, and WQLM was modified and complemented continually.