• Title/Summary/Keyword: p-NPP

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Expression and Purification of the Phosphatase-like Domain of a Voltage-Sensing Phosphatase, Ci-VSP (막 전위 감지 탈인산화 효소, Ci-VSP의 유사 탈인산화 효소 도메인의 발현과 정제)

  • Kim, Sung-Jae;Kim, Hae-Min;Choi, Hoon;Kim, Young-Jun
    • Journal of Life Science
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    • v.21 no.7
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    • pp.1032-1038
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    • 2011
  • Recently identified Ciona intestinalis voltage sensor-containing phosphatase (Ci-VSP) consists of an ion channel-like transmembrane domain (VSD) and a phosphatase-like domain. Ci-VSP senses the change of membrane potential by its VSD and works as a phosphoinositide phosphatase by its phosphatase domain. In this study, we present the construction of His-tagged phosphatase-like domain of Ci-VSP, its recombinant expression and purification, and its enzymatic activity behavior in order to examine the biochemical behavior of phosphatase domain of Ci-VSP without interference. We found that Ci-VSP(248-576)-His can be eluted with an elution buffer containing 25 mM NaCl and 100 mM imidazole during His-tag purification. In addition, we found the proper measurement condition for kinetics study of Ci-VSP(248-576)-His against p-nitrophenyl phosphate (pNPP). We measured the kinetic constant of Ci-VSP(248-576)-His at $37^{\circ}C$, pH 5.0 or 5.5, under 30 min of reaction time, and less than $2.0\;{\mu}g$ of protein amount. With these conditions, we acquired that Ci-VSP(248-576)-His has $K_m$ of $354{\pm}0.143\;{\mu}M$, $V_{max}$ of $0.0607{\pm}0.0137\;{\mu}mol$/min/mg and $k_{cat}$ of $0.359{\pm}0.009751\;min^{-1}$ for pNPP dephosphorylation. Therefore, we produced a pure form of Ci-VSP(248-576)-His, and this showed a higher activity against pNPP. This purified protein will provide the road to a structural investigation on an interesting protein, Ci-VSP.

A Study on Effect of Capture Volume in a Cavity on Direct Containment Heating Phenomena

  • Chung, C.Y.;Kim, M.H.;Lee, H.Y.;Kim, P.S.
    • Nuclear Engineering and Technology
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    • v.28 no.3
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    • pp.290-298
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    • 1996
  • Direct Containment Heating, DCH, is supposed to occur during a core melt-down accident if the primary system pressure is still high at the time of vessel breach in a Nuclear Power Plant (NPP). In this case, DCH is considered to be one of very important severe phenomena during postulated severe accident scenario because of the fast heat transfer rate to atmosphere and the sharp pressure increase in a containment. To reduce the effect of this DCH phenomena, the capture volume wes designed at Ulchin NPP units 3 and 4. But, the effect of this has not been studied extensively. This work consists of experimental and numerical analyses of the effects of capture volume in the cavity on DCH phenomena. The experimental model is a 1/30 scaled-down model of Ulchin NPP units 3 and 4. We used three types of capture volumes to investigate the effect of size. Numerical analysis using CONTAIN 1.2 is performed with the correlation for the dispersed fraction of molten corium from the cavity into the containment derived from the experimental data to examine the effect of capture volume on DCH phenomena in full scale of Ulchin NPP units 3 and 4.

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Study on Seismic Responses for Base Isolated Structure Using Linear 2 DOF System and Its Application for NPP (선형 2자유도계를 이용한 면진구조물의 지진응답 연구 및 원자력발전소 적용)

  • Yoo, Bong;Lee, Jae-Han
    • Proceedings of the Earthquake Engineering Society of Korea Conference
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    • 1997.04a
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    • pp.225-232
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    • 1997
  • A study of effects of design parameters on the seismic responses of base isolated structure is performed to reduce the seismic responses using a linear tw0-degree of freedom system and a lumped-mass model of a nuclear power p;ant(NPP). From the simplified 2 DOF system the optimal isolation frequency being less than 1/10th of the fundamental frequency of superstructure is obtained, and the isolator damping minimizing the peak acceleration depends on superstructure frequency. From the time history analyses for lumped mass model of NPP the optimal damping is calculated as 40% in containment building and 65% in reactor internal structure. Similar results are obtained in 2 DOF system

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Evaluation of Non-Point Pollution Loads in Corn-Autumn Kimchi Cabbage Cultivation Areas by Fertilizer Application Levels Using the APEX Model (APEX 모델을 이용한 옥수수-가을배추 재배지의 시비 수준별 비점오염 부하량 평가)

  • Lee, Jong-Mun;Yeob, So-Jin;Jun, Sang-Min;Lee, Byungmo;Yang, Yerin;Choi, Soon-Kun
    • Journal of The Korean Society of Agricultural Engineers
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    • v.66 no.5
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    • pp.15-27
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    • 2024
  • Agriculture is recognized as an important anthropogenic cause of non-point source loads. Improved understanding of non-point source loads according to fertilization practices can promote climate change and eutrophication mitigation. Thus, this study evaluated the impact of conventional and standard fertilization practices on non-point pollution (NPP) loads in a dual-cropping system, utilizing the Agricultural Policy/Environmental eXtender (APEX) model. Our research objectives were twofold: firstly, to calibrate and validate the APEX model with observed data through experiments from 2018 to 2023; and secondly, to compare the NPP loads under conventional and standard fertilization practices. The model calibration and validation showed satisfactory performance in simulating nitrogen (N) and phosphorus (P) loads, illustrating the model's applicability in a Korean agricultural setting. The simulation results under conventional fertilization practices revealed significantly higher NPP loads compared to the standard fertilization, with P loads under conventional practices being notably higher. Our findings emphasize the crucial role of recommended fertilization practices in reducing non-point source pollution. By providing a quantitative assessment of NPP loads under different fertilization practices, this study contributes valuable information to sustainable nutrient management in agricultural systems facing the dual challenges of climate change and environmental conservation.

Regionality and Variability of Net Primary Productivity and Rice Yield in Korea (우리 나라의 순1차생산력 및 벼 수량의 지역성과 변이성)

  • JUNG YEONG-SANG;BANG JUNG-HO;HAYASHI YOSEI
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.1 no.1
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    • pp.1-11
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    • 1999
  • Rice yield and primary productivity (NPP) are dependent upon the variability of climate and soil. The variability and regionality of the rice yield and net primary productivity were evaluated with the meteorological data collected from Korea Meteorology Administration and the actual rice yield data from the Ministration of Agriculture and Forestry, Korea. The estimated NPP using the three models, dependent upon temperature(NPP-T), precipitation(NPP-P) and net radiation(NPP-R), ranged from 10.87 to 17.52 Mg ha$^{-1}$ with average of 14.69 Mg ha$^{-1}$ in the South Korea and was ranged 6.47 to 15.58 Mg ha$^{-1}$ with average of 12.59 Mg ha$^{-1}$ in the North Korea. The primary limiting factor of NPP in Korea was net radiation, and the secondary limiting factor was temperature. Spectral analysis on the long term change in air temperature in July and August showed periodicity. The short periodicity was 3 to 7 years and the long periodicity was 15 to 43 years. The coefficient of variances, CV, of the rice yield from 1989 to 1998 ranged 3.23 percents to 12.37 percents which were lower than past decades. The CV's in Kangwon and Kyeongbuk were high while that in Chonbuk was the lowest. The prediction model based on th e yield index and yield response to temperature obtain ed from the field crop situation showed reasonable results and thus the spatial distributions of rice yield and predicted yield could be expressed in the maps. The predicted yields was well fitted with the actual yield except Kyungbuk. For better prediction, modification should be made considering radiation factor in further development.

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Evaluation of Corrosion Product Behavior in NPP Secondary System with Complex Amine (복합아민 적용에 따른 원전 2차 계통 부식생성물 거동평가)

  • JUNG, Hyunjun;RHEE, In Hyoung;Kim, Young In
    • Transactions of the Korean Society of Pressure Vessels and Piping
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    • v.10 no.1
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    • pp.96-99
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    • 2014
  • The aim of the study was to evaluate the water treatment of pressurized water reactor secondary side by the mixed amine of ammonia and ethanolamine, from the standpoint of corrosion control, as compared with all volatile treatment of ammonia. The pressurized water reactor systems have switched a secondary side pH control agent to minimize the corrosion in the moisture separator/reheater and feedwater heater systems and the transport of corrosion products into steam generator. As results of field test, pH was increased in the steam generator and the wet steam area of moisture separator/reheater and the concentration of Fe were decreased by more than 50% as compared with water treatment of ammonia.

Substrate Specificity of Alkaline Phosphatase (Alkaline phosphatase의 기질 특이성)

  • ;;E. Waelkens;W. Merlevede
    • YAKHAK HOEJI
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    • v.37 no.6
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    • pp.571-576
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    • 1993
  • The substrate specificity of the purified rabbit plasma alkaline phosphatase (ALPase) was determined towards a extended range of potential substrates including relatively simple phosphate derivatives as p-NPP and indolyl phosphate, and several synthetic peptides and phosphoproteins. These results further estabilish the broad substrate specificity of these circulating enzymes. Interestingly, the plasma ALPase preferentially dephosphorylates Thr over Ser residues, as demonstrated with a series of synthetic peptides. The latter result is in contradiction to the behaviour of the tissue ALPase, which is thought to the ultimate source of plasma ALPase, and open therefore new perspectives with respective to the origin and "solubilisation" processes of these enzymes. Dephsphrylation of protein substrates by endogenous and isolated plasma ALPases indicates that ALPase probably displays protein phosphatase activity in vivo.

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Efficacy of Supplemental Microbial Phytase on Laying Performance and Phosphorus Utilization II. Effect of Microbial Phytase at Different Phosphorus Levels and High Calcium Content on Laying Performance and Phosphorus Utilization (산란생산성과 인 이용성에 대한 Microbial Phytase의 첨가 효과 II. 무기태인 수주닝 다르고 칼슘수준이 높은 사료에 Microbial Phytase 첨가가 산란성 및 인 이용성에 미치는 영향)

  • 김상호;유동조;이상진;강보석;서옥석;최철환;이원준;류경선
    • Korean Journal of Poultry Science
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    • v.27 no.1
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    • pp.25-30
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    • 2000
  • Present study was conducted to investigate effects of microbial phytase in laying hen diets on utilization of non-phytate phosphorus(NPP) whose levels were adjusted to be adequate or lower than that of NRC requirements. Birds of control roup were fed a diet containing 0.275% NPP and 3.4% Ca, satisfying the NRC(1994) feeding standard. bird on T1, T2 and T3 were allowed to eat diets containing NPP at 100, 80 and 60%, respectively, of Control group, and 4.0% Ca level along with a microbial phytase added at a level of 300 DPU. Three hundred and sixty, ISA Brown layers, 23-week-old, divided into four treatment groups with three replications per treatment and 30 layers per replication were fed the diets for 12 weeks. Levels of feed intake were not different among the groups, The egg mass/feed intake ratio appeared better in T2 group by about 8%, though without a statistical significance, compared to that of control. Egg production fate tended to be improved over the control group by feeding the 100%(T1) and 80%(T2) NPP diets added with phytase, with a significant difference for T2(p<0.05). Mean egg weight and egg shell quality, measured by breaking strength and thickness of the egg shell, of the T2 group tended to show numerically better, without a significance than those of control. Furthermore, birds of the T2 group showed higher calcium and phosphorus contents in tibia by about 9%(p<0.05) than the control. Overall performances of birds in T1 appeared better than those of control, but tended to be lower than those of the T2. The birds in T3 performed similar to the those of the other dietary groups except the relatively low tibia calcium level. In conclusion, the results of this study suggest that supplementation of microbial phytase at a level of 300 DPU was effective to spare about 20% of NPP in laying hen diets without any adverse effects on production performances and bone quality.

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General Enzymatic Properties of Human Histidine Acid Phosphatase-Phytase (히스티딘 에시드 포스파테이즈(Histidine Acid Phosphatase) 계열 인간 파이테이즈(Phytase)의 일반적 특성규명)

  • Cho, Jaie-Soon
    • Journal of Animal Science and Technology
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    • v.51 no.2
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    • pp.177-182
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    • 2009
  • The glycosylated human MINPP (multiple inositol polyphosphate phosphatase), which was recombinantly over-expressed by using industrial host, Pichia pastoris, showed the phytase activity against phytate ($InsP_6$) and the enzyme activity of the unglycosylated counterpart was decreased to 30%. The optimal phytase activity occurred at pH 7.4. The human MINPP showed high substrate specificity for $InsP_6$ with little activity on other organic phosphate conjugates such as para-nitrophenylphosphate (pNPP), ATP, and ribose-1-phosphate (R-1-P). The phosphatase activity against 2,3-bisphosphoglycerate (2,3-BPG) by human MINPP was increased to 1.2-fold in the presence of stimulator, 1 mM 2-phosphoglycolate (2-PG) but the phytase activity against $InsP_6$ was not affected by addition of 1 mM 2-PG. The phosphatase activity against 2,3-BPG by human MINPP was not increased in the presence of 2 mM $Mg^{2+}$ or 100 mM $Cl^-$.

Purification and Characterization of Protein Phosphatase 2A from Petals of the Tulip Tulipa gesnerina

  • Azad, Md. Abul Kalam;Sawa, Yoshihiro;Ishikawa, Takahiro;Shibata, Hitoshi
    • BMB Reports
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    • v.39 no.6
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    • pp.671-676
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    • 2006
  • The holoenzyme of protein phosphatase (PP) from tulip petals was purified by using hydrophobic interaction, anion exchange and microcystin affinity chromatography to analyze activity towards p-nitrophenyl phosphate (p-NPP). The catalytic subunit of PP was released from its endogenous regulatory subunits by ethanol precipitation and further purified. Both preparations were characterized by immunological and biochemical approaches to be PP2A. On SDS-PAGE, the final purified holoenzyme preparation showed three protein bands estimated at 38, 65, and 75 kDa while the free catalytic subunit preparation showed only the 38 kDa protein. In both preparations, the 38 kDa protein was identified immunologically as the catalytic subunit of PP2A by using a monoclonal antibody against the PP2A catalytic subunit. The final 623- and 748-fold purified holoenzyme and the free catalytic preparations, respectively, exhibited high sensitivity to inhibition by 1 nM okadaic acid when activity was measured with p-NPP. The holoenzyme displayed higher stimulation in the presence of ammonium sulfate than the free catalytic subunit did by protamine, thereby suggesting different enzymatic behaviors.