• 제목/요약/키워드: AGP

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A truncated form of human alpha 1-acid glycoprotein is useful as a molecular tool for insect glycobiology

  • Morokuma, Daisuke;Hino, Masato;Tsuchioka, Miho;Masuda, Akitsu;Mon, Hiroaki;Fujiyama, Kazuhito;Kajiura, Hiroyuki;Kusakabe, Takahiro;Lee, Jae Man
    • International Journal of Industrial Entomology and Biomaterials
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    • v.36 no.1
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    • pp.15-24
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    • 2018
  • N-glycosylation is an important posttranslational modification that results in a variety of biological activities, structural stability, and protein-protein interactions. There are still many mysteries in the structure and function of N-glycans, and detailed elucidation is necessary. Baculovirus expression system (BES) is widely used to produce recombinant glycoproteins, but it is not suitable for clinical use due to differences in N-glycan structure between insects and mammals. It is necessary to develop adequate model glycoproteins for analysis to efficiently alter the insect-type N-glycosylation pathway to human type. The previous research shows the recombinant alpha 1-acid glycoprotein (${\alpha}1AGP$) secreted from silkworm cultured cells or larvae is highly glycosylated and expected to be an excellent research candidate for the glycoprotein analysis expressed by BES. Therefore, we improved the ${\alpha}1AGP$ to be a better model for studying glycosylation. The modified ${\alpha}1AGP$ (${\alpha}1AGP{\Delta}$) recombinant protein was successfully expressed and purified by using BES, however, the expression level in silkworm cultured cells and larvae were lower than that of the ${\alpha}1AGP$. Subsequently, we confirmed the detailed profile of N-glycan on the ${\alpha}1AGP{\Delta}$ by LS/MS analysis the N-glycan structure at each glycosylation site. These results indicated that the recombinant ${\alpha}1AGP{\Delta}$ could be usable as a better model glycoprotein of N-glycosylation research in BES.

Molecular Modification of Perilla Lipid Composition

  • Hwang, Young-Soo;Kim, Kyung-Hwan;Hwang, Seon-Kap;Lee, Sun-Hwa;Lee, Seong-Kon;Kim, Jung-Bong;Park, Sang-Bong;Tom Okita;Kim, Donghern
    • Journal of Plant Biotechnology
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    • v.1 no.1
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    • pp.20-30
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    • 1999
  • In order to modify lipid production of Perilla qualitatively as well as quantitatively by genetic engineering, genes involved in carbon metabolism were isolated and characterized. These include acyl-ACP thioesterases from Perilla frutescens and Iris sp., four different $\beta$-ketoacyl- ACP synthases from Perilla frutescens, and two $\Delta$15 a-cyl-ACP desaturases(Pffad7, pffad3). Δ15 acyl-ACP desa turase (Δ15-DES) is responsible for the conversion of linoleic acid (18:2) to $\alpha$-linolenic acid (ALA, 18:3). pffad 3 encodes Δ15 acyl-desaturase which is localized in ER membrane. On the other hand, Pffad7 encodes a 50 kD plastid protein (438 residues), which showed highest sequence similarity to Sesamum indicum fad7 protein. Northern blot analysis revealed that the Pffad7 is highly expressed in leaves but not in roots and seeds. And Pffad3 is expressed throughout the seed developmental stage except very early and fully mature stage. We constructed Pffad7 gene under 355 promoter and Pffad3 gene under seed specific vicillin promoter. Using Pffad7 construct, Perilla, an oil seed crop in Korea, was transformed by Agrobacterium leaf disc method. $\alpha$-linolenic acid contents increased in leaves but decreased in seeds of transgenic Perilla. Currently, we are transforming Perilla with Pffad3 construct to change Perilla seed oil composition. We isolated three ADP-glucose pyrophosphorylase (AGP) genes from Perilla immature seed specific cDNA library. Nucleotide sequence analysis showed that two of three AGP (Psagpl, Psagp2) genes encode AGP small subunit polypeptides and the remaining (Plagp) encodes an AGP large subunit. PSAGPs, AGP small subunit peptide, form active heterotetramers with potato AGP large subunit in E. coli expressing plant AGP genes.

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Effects of Antibiotic Growth Promoter and Characterization of Ecological Succession in Swine Gut Microbiota

  • Unno, Tatsuya;Kim, Jungman;Guevarra, Robin B.;Nguyen, Son G.
    • Journal of Microbiology and Biotechnology
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    • v.25 no.4
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    • pp.431-438
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    • 2015
  • Ever since the ban on antibiotic growth promoters (AGPs), the livestock death rate has increased owing to pathogenic bacterial infections. There is a need of developing AGP alternatives; however, the mechanisms by which AGP enhances livestock growth performance are not clearly understood. In this study, we fed 3-week-old swine for 9 weeks with and without AGPs containing chlortetracycline, sulfathiazole, and penicillin to investigate the effects of AGPs on swine gut microbiota. Microbial community analysis was done based on bacterial 16S rRNA genes using MiSeq. The use of AGP showed no growth promoting effect, but inhibited the growth of potential pathogens during the early growth stage. Our results showed the significant increase in species richness after the stabilization of gut microbiota during the post-weaning period (4-week-old). Moreover, the swine gut microbiota was divided into four clusters based on the distribution of operational taxonomic units, which was significantly correlated to the swine weight regardless of AGP treatments. Taxonomic abundance analysis indicated a negative correlation between host weight and the abundance of the family Prevotellaceae species, but showed positive correlation to the abundance of the family Spirochaetaceae, Clostridiaceae_1, and Peptostreptococcaeae species. Although no growth performance enhancement was observed, the use of AGP inhibited the potential pathogens in the early growth stage of swine. In addition, our results indicated the ecological succession of swine gut microbiota according to swine weight. Here, we present a characterization of swine gut microbiota with respect to the effects of AGPs on growth performance.

Comparison of Algal Growth Potentials in the Large Reservoirs and River Mainstream of Naktong River Watershed (낙동강 수계 대형 인공호 및 하천본류의 조류성장 잠재력 비교)

  • You, Kyung-A;Shin, Jae-Ki;Hwang, Soon-Jin
    • Korean Journal of Ecology and Environment
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    • v.39 no.1 s.115
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    • pp.138-144
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    • 2006
  • Algal growth potential test (AGPT) has been used as a tool for assessing biological productivity potential in the aquatic ecosystems. This study was conducted to compare the productivity potentials of large reservoirs (Lakes Andong, Hapchon and Jinyang, and Naktong estuarine dam) and river sites (Sangju, Koryung, and Samlangjin) located in the Naktong River watershed. AGPT was conducted in both non-monsoon and mosoon season (February, April, July and September) of 2003, using Microcystis aeruginosa as a test alga. The AGPs in the reservoirs were relatively much lower than those of river sites. The river AGPs increased towards upstream close to the influent streams, while it generally decreased towards downstream. Immediately after the abrupt increase in influent discharge in summer, the AGP became similar between midstream and downstream sites. The water quality of river and reservoirs deteriorated during the drought period in accordance with AGP: it was the highest during this period. The AGPs showed the closest correlation with the P concentration, leading to the conclusion that bioavailable P is highly influential to the algal growth in both lentic and lotic ecosystems in the Naktong River watershed. Based on the AGPs, the water quality of tested sites was likely eutrophic. Our results suggest that AGPT be a useful tool in evaluating the productivity potential and trophic state of the water body as well as determining the nutrients that limit the growth of algae.

PRODUCT10N OF KSR-III AIRGLOW PHOTOMETERS TO MEASURE MUV AIRGLOWS OF THE UPPER ATMOSPHERE ABOVE THE KOREAN PENINSULAR (한반도 상공의 고층대기 중간 자외선 대기광 측정을 위한 KSR-III 대기광도계 제작)

  • Oh, T.H.;Park, K.C.;Kim, Y.H.;Yi, Y.;Kim, J.
    • Journal of Astronomy and Space Sciences
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    • v.19 no.4
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    • pp.305-318
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    • 2002
  • We have constructed two flight models of airglow photometer system (AGP) to be onboard Korea Sounding Rocket-III (KSR-III) for detection of MUV dayglow above the Korean peninsular. The AGP system is designed to detect dayglow emissions of OI 2972${\AA}$, $N_2$ VK(0,6) 2780${\AA}$, $N_2$ 2PG 3150${\AA}$ and background 3070${\AA}$ toward the horizon at altitudes between 100 km and 300 km. The AGP system consists of a photometer body, a baffle an electronic control unit and a battery unit. The MUV dayglow emissions enter through a narrow band interference filter and focusing lens of the photometer, which contains an ultraviolet sensitive photomultiplier tube. The photometer is equipped with an in-flight calibration light source on a circular plane that will rotate at the rocket's apogee. A bane tube is installed at the entry of the photometer in order to block strong scattering lights from the lower atmosphere. We have carried out laboratory measurements of sensitivity and in-flight calibration light source for the AGP flight models. Although absolute sensitivities of the AGP flight models could not be determined in the country, relative sensitivities among channels are well measured so that observation data during rocket flight in the future can be analyzed with confidence.

Effect of the Sewage and Wastewater Plant Effluent on the Algal Growth Potential in the Nakdong River Basin (낙동강 수계 하.폐수 처리시설의 방류수가 조류 성장 잠재력에 미치는 영향)

  • Seo,Jeong-Gwan;Lee,Jae-Jeong;Yang,Sang-Yong;Jeong,Ik-Gyo
    • ALGAE
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    • v.18 no.2
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    • pp.157-167
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    • 2003
  • Effect of the effluent of the sewage and wastewater plants on the algal growth was investigated from the 19 plants located in the Nakdong river basin. Most of the samples showed high values of the algal growth potential (AGP) when they were mixed with natural river water at 20% of final concentration. At 20% of the mixing ratio, the mixed effluents of sewage and wastewater showed 3.5 and 1.8 times higher AGP than those of the natural river water. The higher AGP values are attributable to the high contents of phosphorus and ammonium in the effluent. The mixing ratio of effluents of the discharge/river flow was highest in the Kumho River (42.8%) followed by the middle of Nakdong River (22.7%), Kam Stream (13.9%), Byungsung Stream (13.3%), Yangsan Stream (7.9%), and Young River (5.4%). Comparison of the trophic state of the effluents with natural river water indicated that the effluents showed higher trophic values than natural water. Concentrations of total phosphorus, total nitrogen and conductivity in the effluents were 12.3, 4.9 and 5.3 times higher than the those found in natural river water respectively. The AGP values were highly related with the trophicity of the water especially on the concentrations of phosphate and ammonium. Toxicities of the treated sewage water, wastewater and livestock waste water tested by the luminescent bacteria, Vibrio fischerii were generally low.

Evaluation of Water Quality after Rehabilitation of Cheonggye Stream using AGP Test (조류성장잠재력 조사를 이용한 청계천 복원 이후 수질 평가)

  • Park, Myung-Hwan;Hwang, Soon-Jin;Suh, Mi-Yeon;Kim, Yong-Jae;Kim, Baik-Ho
    • Korean Journal of Ecology and Environment
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    • v.40 no.2
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    • pp.234-243
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    • 2007
  • Algal growth potential (AGP) test was performed to evaluate the water quality and changes in phytoplankton communities before and after a heavy rain event at six sampling sites in Cheonggye Stream (St. 1 and 2), Jungnang Stream (St. 3 and 4), and Lower part of Han River System (St. 5 and 6) after rehabilitation of Cheonggye Stream, October 2005. To test AGP on each sampling site, cyanobacterium Microcystis aeruginosa was applied as a standard alga. Total nitrogen (TN) showed high values at Jungnang Stream, while St. 4 recorded highest values in this study. However, TN values of Cheonggye Stream and Lower Part of Han River showed similar levels. Total phosphate (TP) also showed high values at Jungnang Stream, while St. 4 recorded highest. However, TP in Cheonggye Stream were extremely low levels. Although chlorophyll-${\alpha}$ (chi-${\alpha}$) contents before the rain event were similar through the sampling sites, chl-${\alpha}$ after the rain increased dramatically at Jungnang Stream and Lower part of Ban River. In particular, after the rain, TP was the highest at St. 4, where Cyclotella sp. dominated the phytoplankton community. When compared with control, AGP values before the rain were comparatively low in all sites, while those after the rain highly increased with the dose-dependently of field water added, due perhaps to the increased nutrients by rainfall. Similar results were observed in Cheonggye Stream. Therefore, for the aesthetic fostering for the citizens, although Cheonggye Stream was Presently being sustained by treated water supply, they have a potential of outbreak of phytoplankton by the increased nutrients supply when a heavy rain comes.

Dynamic Patterns of Systemic Innate Immunity and Inflammatory Associated Factors in Experimental Caprine Coccidiosis

  • Tadayon, Shabnam;Razavi, Seyed Mostafa;Nazifi, Saeed
    • Parasites, Hosts and Diseases
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    • v.54 no.6
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    • pp.719-724
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    • 2016
  • The present study was designed to assess the dynamic patterns of pro-inflammatory cytokines, including $IFN-{\gamma}$, $TNF-{\alpha}$, IL-4, IL-6, acute phase protein (${\alpha}1$-acid-glycoprotein, AGP), and an inflammation associated factor (adenosine deaminase; ADA) following experimental caprine coccidiosis. Ten kids aging from 2 to 4 months were infected orally with $5{\times}10^4$ sporulated oocysts and 10 animals served as controls. Blood samples were collected in both groups before infection and at days 3, 7, 14, 21, 28, and 35 post-infection (PI), and the levels of above-mentioned factors were measured. $IFN-{\gamma}$, $TNF-{\alpha}$, IL-4, IL-6, AGP, and ADA activities were significantly higher in infected animals from day 7 PI (P<0.05). In conclusion, the circulatory levels of most systemic inflammatory markers, including pro-inflammatory cytokines ($IFN-{\gamma}$, $TNF-{\alpha}$, IL-4, IL-6), AGP, and ADA increased significantly starting from day 3 to day 7 PI in caprine coccidiosis.

Gene Cloning, Expression, and Characterization of Glucose-1-Phosphatase from Enterobacter cloacae B11

  • Kim, Young-Ok;Park, In-Suk;Nam, Bo-Hye;Kong, Hee-Jeong;Kim, Woo-Jin;Lee, Sang-Jun;Kim, Kyung-Kil
    • Fisheries and Aquatic Sciences
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    • v.13 no.1
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    • pp.49-55
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    • 2010
  • A bacterial strain with phytase and glucose-1-phosphatase activity was isolated from seawater. The colony was identified as an Enterobacter cloacae strain and named E. cloacae B11. A gene, agpEnB11, coding for an intracellular acid glucose phosphatase was cloned from the strain and sequenced. It comprised 1,242 nucleotides and encoded a polypeptide of 413 amino acids. Recombinant glucose-1-phosphatase (AgpEn) was overexpressed in Escherichia coli and purified using Ni-NTA column under native conditions. Purified protein displayed a single band of 47 kDa on SDS-PAGE. AgpEn hydrolyzed a wide variety of phosphorylated compounds, with high activity for glucose-1-phosphate and glucose-6-phosphate. Optimum pH and temperature for enzyme activity were pH 5.0 and $50^{\circ}C$, respectively. Enzyme activity was stimulated by $Ca^{2+}$ and $Co^{2+}$, and inhibited by $Cu^{2+}$.

Cyanobacterial Bioassay (AGP test) on the Water Fertility of Treated Wastewater Effluents Discharged into Euiam and Paldang Reservoirs, Korea (의암호 및 팔당호에 유입되는 주요 하수처리 방류수의 수질 비옥도 생물검정: Algal Growth Potential(AGP) Test)

  • Seo, Wanbum;Lee, Su-Woong;Kim, Keonhee;Park, Chaehong;Choi, Bong-Geun;Sim, Yeon Bo;Hwang, Soon-Jin;Shin, Jae-Ki
    • Korean Journal of Ecology and Environment
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    • v.50 no.1
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    • pp.79-95
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    • 2017
  • Euiam and Paldang Reservoirs have often been facing water quality problems, such as eutrophication, algal blooms and off-flavors by treated wastewater effluent (TWE) in the North-Han and the Han River basins, but little is examined on the direct biological effect of TWE. This study tested algal growth potential (AGP) of four TWEs discharged into Euiam and Paldang Reservoirs to evaluate water fertility in September 2014 and March and September 2015. Test alga was used Anabaena circinalis isolated from Paldang Reservoir. Mean concentration of T-N and T-P in TWEs was $3,956.7{\mu}g\;N\;L^{-1}$ and $50.8{\mu}g\;P\;L^{-1}$, and the proportion of $NO_3-N$ and $PO_4-P$ to the total fraction was 72.1% and 40.8%, respectively. Both N and P were high in TWEs, but much higher N than P concentration indicates strong P-limitation. As a consequence, the maximum AGP was determined by $PO_4-P$ concentration (r=0.998, p<0.01). Mean AGP value was $15.4mg\;dw\;L^{-1}$ among four effluents indicating its eutrophic condition. Due to the establishment of tertiary (advanced T-P) treatment method in the studied plants recently, P concentration was significantly decreased in TWEs compared to the years prior to 2012. However, P concentration seems to be still high enough to cause eutrophication and algal blooms. Therefore, wastewater treatment to P-free level needs to be considered if effluents are directly discharged into the drinking water resources.