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Isolation and Characterization of a Novel Calcium/Calmodulin-Dependent Protein Kinase, AtCK, from Arabidopsis

  • Jeong, Jae Cheol;Shin, Dongjin;Lee, Jiyoung;Kang, Chang Ho;Baek, Dongwon;Cho, Moo Je;Kim, Min Chul;Yun, Dae-Jin
    • Molecules and Cells
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    • v.24 no.2
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    • pp.276-282
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    • 2007
  • Protein phosphorylation is one of the major mechanisms by which eukaryotic cells transduce extracellular signals into intracellular responses. Calcium/calmodulin ($Ca^{2+}/CaM$)-dependent protein phosphorylation has been implicated in various cellular processes, yet little is known about $Ca^{2+}/CaM$-dependent protein kinases (CaMKs) in plants. From an Arabidopsis expression library screen using a horseradish peroxidase-conjugated soybean calmodulin isoform (SCaM-1) as a probe, we isolated a full-length cDNA clone that encodes AtCK (Arabidopsis thaliana calcium/calmodulin-dependent protein kinase). The predicted structure of AtCK contains a serine/threonine protein kinase catalytic domain followed by a putative calmodulin-binding domain and a putative $Ca^{2+}$-binding domain. Recombinant AtCK was expressed in E. coli and bound to calmodulin in a $Ca^{2+}$-dependent manner. The ability of CaM to bind to AtCK was confirmed by gel mobility shift and competition assays. AtCK exhibited its highest levels of autophosphorylation in the presence of 3 mM $Mn^{2+}$. The phosphorylation of myelin basic protein (MBP) by AtCK was enhanced when AtCK was under the control of calcium-bound CaM, as previously observed for other $Ca^{2+}/CaM$-dependent protein kinases. In contrast to maize and tobacco CCaMKs (calcium and $Ca^{2+}/CaM$-dependent protein kinase), increasing the concentration of calmodulin to more than $3{\mu}M$ suppressed the phosphorylation activity of AtCK. Taken together our results indicate that AtCK is a novel Arabidopsis $Ca^{2+}/CaM$-dependent protein kinase which is presumably involved in CaM-mediated signaling.

Nutritional Compositions and Physicochemical Properties of Two Domestic Aronia (A. melanocarpa) Varieties (국내산 아로니아 두 품종(네로, 바이킹)의 영양성분 및 생리활성 분석)

  • Jeong, Yeon-Jeong;Lee, A Reum;Park, Jae-Ho;Kim, Youngho;Kwon, Yeuseok;Hong, Eui Yon;Han, Nam Soo;Eom, Hyun-Ju
    • The Korean Journal of Food And Nutrition
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    • v.29 no.2
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    • pp.283-289
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    • 2016
  • The nutritional compositions and physicochemical properties of two aronia varieties (Viking and Nero) cultivated in Chungbuk were investigated. The proximate compositions of Nero and Viking powder were:13% moisture, 0.58% and 0.6% ash, 0.73% and 0.33% crude lipid, 5.47% and 6.51% crude protein, 15.65% and 16.15% crude dietary fiber, and 64.36 and 64.43% crude carbohydrate, respectively. The proximate contents of the aronia samples were not significantly different. Among the physicochemical properties, there was also no significant change in total anthocyanin and antioxidant activity. However, Viking had higher total polyphenol contents and ${\alpha}$-glucosidase inhibition activity than Nero. The mineral contents of Al, Fe, Mn, Mg, Ca, Na, K, Cu and Zn in the two aronias were determined. Cu and Zn were not detected in both varieties. With regard to Al, Na and K, Nero was higher than Viking, while Ca was higher in Viking than Nero. From the results, Viking could be suggested to be beneficial for food processing.

Preparation of PVdF Composite Nanofiber Membrane by Using Manganese-Iron Oxide and Characterization of its Arsenic Removal (망간-철 산화물을 이용한 PVdF 나노섬유복합막의 제조 및 비소 제거 특성 평가)

  • Yun, Jaehan;Jang, Wongi;Park, Yeji;Lee, Junghun;Byun, Hongsik
    • Membrane Journal
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    • v.26 no.2
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    • pp.116-125
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    • 2016
  • This study described a synthesis of MF having a arsenic removal characteristics and the fundamental research was performed about the simultaneous removal system of both As(III) and As(V) ions with the composite nanofiber membrane (PMF) based on PVdF and MF materials for the water-treatment application. From the TEM analysis, the shape and structure of MF materials was investigated. The mechanical strength, pore-size, contact angle and water-flux analysis for the PMF was performed to investigate the possibility of utilizing as a water treatment membrane. From these results, the PMF11 showed the highest value of mechanical strength ($232.7kgf/cm^2$) and the pore-diameter of composite membrane was reduced by introducing the MF materials. In particular, their pore diameter decreased with an increase of iron oxide composition ratio. The water flux value of PMF was improved about 10 to 60% compared with that of neat PVdF nanofiber membranes. From the arsenic removal characterization of prepared MF materials and PMF, it was shown the simultaneous removal characteristics of both As(III) and (V) ions, and the MF01, in particular, showed the highest adsorption-removal rate of 93% As(III) and 68% As(V), respectively. From these results, prepared MF materials and PMF have shown a great potential to be utilized for the fundamental study to improve the functionality of water treatment membrane.

Chemical Composition and Features of Asian Dust Observed in Korea (2000~2002) (2000~2002년 우리나라에서 관측된 황사의 화학 조성 및 특성)

  • Shin S.A;Han J.S;Hong Y.D;Ahn J.Y;Moon K.J;Lee S.J;Kim S.D
    • Journal of Korean Society for Atmospheric Environment
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    • v.21 no.1
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    • pp.119-129
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    • 2005
  • The ambient TSP data measured at Seoul, Incheon. Taean, Daegu, Busan in Korea were used to explain the chemical composition and general features of Asian Dust (AD) observed in Korea. 9 episodes out of 19 were sampled from 2000 through May 2002, and measurements were conducted covering ionic and metal components with mass concentration. The results showed that daily averaged mass concentration (TSP) during the AD episodes was 458 $\mu\textrm{g}$/㎥, and ionic and metal concentrations were 27.93 $\mu\textrm{g}$/㎥ and 71.7 $\mu\textrm{g}$/㎥, respectively, accounting for 6.1 % and 15.5% of the total aerosol mass. TSP concentrations during episodes were varied from 120 to 1742 $\mu\textrm{g}$/㎥ according to the impact of Asian Dusts and had a tendency of showing higher values at sites in the west side of Korea, which can be explained by the effect of diffusion and deposition. In this study, ionic components like Ca (NO$_3$)$_2$, CaSO$_4$, NaNO$_3$, Na$_2$SO$_4$ were prominent types in secondary aerosol during AD periods and also indicated that V, Co as well as soil elements such as Ca, Fe, Mg, Mn, K correlated well with Al, while Cu, Cd, Pb, Zn didn't agree well with it. In addition, enrichment factors (EFs) for each metal component were obtained to provide simple information about source contribution of Asian Dust, and the results were compared with those from other AD studies. In this study, the results showed that aerosol properties in Korea during the Asian Dust were considerably different from those of general atmospheric condition and specially varied from case to case rather than site to site, which implies that there are certain variations in the soil of source region, pathways of air mass, and meteorological condition. For the enhanced study, those factors should be combined with the features of Asian Dust resolved from this study.

Geochemical Characteristics of the Continental Shelf and Slope Sediments off the Southeastern Coast of Korea (한국 동남해역 대륙붕과 대륙사면 표층퇴적물의 지화학적 특성)

  • Lee, Chang-Bok;Park, Yong-Ahn;Kang, Hyo-Jin;Kim, Dae-Chul
    • The Korean Journal of Quaternary Research
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    • v.5 no.1
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    • pp.15-31
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    • 1991
  • A total of 90 surface sediment samples, collected from the continental margin area bordering east and southeast coast of Korea, were subject to the geochemical analyses with the aim of filling the gap in our knowledge of this environment. The analyzed items included the major elements (Al, Fe, Mg, Ca, K, Na, p and Mn), organic carbon, and some trace metals (Ba, Co, Cu, Sr and Zn). The sediment grain-size exerted a predominant influence on the contents of most elements, with the exceptions of Ca, K, Sr and Ba. The Ca and Sr contents, being closely interrelated each other, were mainly controlled by the calcium carbonate content. The K content, on the other hand, appeared to be influenced by both illite and feldspar. The Ba content showed a certain relationship with that of K, suggesting a common source of these two elements; potassium feldspar. The R-mode factor analysis result also reaffirmed the above-mentioned controlling factors on the sediment geochemistry. The grain-size dependency of trace metals obscures their areal distribution pattern from the total contents. However, with the metal/aluminum ratios we could differentiate the subtle difference in the metal enrichment. Hence, sediments of the southern coastal area appear to receive some anthropogenic inputs of metals, though the effect is still negligible.

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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.

Refractory Textile Wastewater Treatment Using Cell-Immobilized Polyethylene glycol Media (PEG 포괄고정화담체를 이용한 난분해성 염색폐수 처리)

  • Han, Duk-Gyu;Cho, Young-Jin;Bae, Woo-Keun;Hwang, Byung-Ho;Lee, Yong-Woo
    • Journal of Korean Society of Environmental Engineers
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    • v.28 no.3
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    • pp.345-350
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    • 2006
  • This study investigated the removal of recalcitrant organics in dyeing wastewater using a fluidized bed reactor(FBR) that contained cell-immobilized pellets. The pellets were manufactured and condensing the gel phase by mixing PEG-polymer and cells to form micro-porous PEG-polymer pellets whose size were ${\Phi}\;4mm{\times}H\;4mm$ on average. An industrial activated sludge without any pre-adaptation was used for the cell immobilization because it gave an equivalent removal efficiency to a pre-adapted sludges. The feed was obtained from an effluent of a biological treatment plant, which contained $SCOD_{Cr}$ of 330 mg/L and $SBOD_5$ of 20 mg/L. The $SCOD_{Cr}$ removal efficiency was over 45% and the effluent $COD_{Mn}$ concentration was less than 100 mg/L at HRTs from 6 to 24 hrs. The optimum HRT in the FBR was determined as 12 hrs considering the removal efficiency and cost. When a raw wastewater containing 768 mg/L of $COD_{Cr}$ was fed to the FBR, the effluent $COD_{Cr}$ concentration increased only slightly, giving a 70% of $COD_{Cr}$ removal or a 97% of $BCOD_5$ removal. This indicated that the FBR had an excellent capability of biodegradable organics removal also. In conclusion, the FBR could be applied to textile wastewater treatment in place of an activated sludge process.

Effect of the Physicochemical Properties of Soil on the Arsenic Bioaccessibility (비소용출에 대한 토양의 물리화학적 특성 영향)

  • Yang, Jae-Kyu;Chang, Yoon-Young
    • Journal of Korean Society of Environmental Engineers
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    • v.28 no.7
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    • pp.731-737
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    • 2006
  • Four well-characterized soils collected from A- and B-horizon in the Department of Energy Oak Ridge Reservation in USA, mainly distributed with Inceptisol(Inc) and Ultisol(Ult) soils, were used in this work. The bioaccessibility of arsenic as well as oxidation phenomena of As(III) was investigated with soils spiked with As(III) and As(V) using a physiologically based extraction test(PBET) at pH 1.5 and 1:100 soil to solution ratio. Also effect of aging time on the bioaccessibility of arsenic was investigated over the 6 months. After 48 hours(fresh) contacting As(V) solution with soils, all soils rapidly and strongly sequestrated As(V), especially Ult-B. However, little sequestration was observed after 3-months. When As(III) was spiked on the same soils, a great portion of As(III) was oxidized to As(V) after 48 hrs, especially Inc-A and Ult-A soils, which is strongly related with Mn content in soils. By using As(III)-spiked soils, much reduced bioaccessibility as total arsenic was observed from Inc-B and Ult-B soils over the 6 months aging time compared to that from Inc-A and Ult-A soils. This result can be explained by the continuous sequestration of As(V), produced from oxidation of As(III), onto Inc-B and Ult-B soils having much amount of iron. The trend of As(III) sequestration over six months aging time was quite similar with that of As(V) sequestration.

Step by Step Analysis of Root Canal Instrumentation with ProTaper® (ProTaper를 이용한 근관 형성의 단계별 분석)

  • Kim, Mi-Hee;Huh, Bock;Kim, Hyeon-Cheol;Park, Jeong-Kil
    • Restorative Dentistry and Endodontics
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    • v.31 no.1
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    • pp.50-57
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    • 2006
  • The purpose of this study was to investigate influence of each file step of $ProTaper^{(R)}$ system on canal transportation. Twenty simulated canals were prepared with either engine-driven $ProTaper^{(R)}$ or manual $ProTaper^{(R)}$, Group R-resin blocks were instrumented with rotary $ProTaper^{(R)}$ and group M-resin blocks were instrumented with manual $ProTaper^{(R)}$. Pre-operative resin blocks and post-operative resin blocks after each file step preparation were scanned. Original canal image and the image after using each file step were superimposed for calculation of centering ratio The image after using each file step alld image after using previous file step were superimposed for calculation of the amount of deviation. Measurements were taken horizontally at five different levels (1 2, 3, 4 and 5 mm) from the level of apical foramen. In rotary $ProTaper^{(R)}$ instrumentation group, centering ratio and the amount of deviation of each step at all levels were not significantly different (p>0.05). In manual $ProTaper^{(R)}$ instrumentation group, centering ratio and the amount of deviation of each step at all levels except of 1 mm were not significantly different (p>0.05). At the level of 1 mn, F2 file step had significantly large centering ratio and the amount of deviation (p<0.05). Under the condition of this study, F2 file step of manual ProTaper tended to transport the apical part of the canals than that of rotary $ProTaper^{(R)}$.

Shear bond strength of dentin bonding agents cured with a plasma arc curing light (전단접착강도와 관련된 Plasma Arc Curing Light의 중합효율평가)

  • Kwon, Young-Chul;Kim, Sun-Young;Chung, Sae-Joon;Han, Young-Chul;Lee, In-Bog;Son, Ho-Hyun;Um, Chung-Moon;Cho, Byeong-Hoon
    • Restorative Dentistry and Endodontics
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    • v.33 no.3
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    • pp.213-223
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    • 2008
  • The objective of this study was to compare dentin shear bond strength (DSBS) of dentin bonding agents (DBAs) cured with a plasma arc (PAC) light curing unit (LCU) and those cured with a light emitting diode (LED) LCU. Optical properties were also analyzed for Elipar freelight 2 (3M ESPE); LED LCU, Apollo 95E (DMT Systems); PAC LCU and VIP Junior (Bisco); Halogen LCU. The DBAs used for DSBS test were Scotchbond Multipurpose (3M ESPE), Singlebond 2 (3M ESPE) and Clearfil SE Bond (Kuraray). After DSBS testing, fractured specimens were analyzed for failure modes with SEM. The total irradiance and irradiance between 450 nm and 490 nm of the LCUs were different. LED LCU showed narrow spectral distribution around its peak at 462 nm whereas PAC and Halogen LCU showed a broad spectrum. There were no significant differences in mean shear bond strength among different LCUs (P > 0.05) but were significant differences among different DBAs (P < 0.001)