• Title/Summary/Keyword: GLC

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Comparison of Properties of Polyclonal Anti-N-Acetylchitooligosaccharides and Anti-Chitooligosaccharides Antibodies Produced for ELISA

  • Shim, Youn-Young;Shon, Dong-Hwa;Kwak, Bo-Yeon;Yu, Jae-Hoon;Chee, Kew-Mahn
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.686-692
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    • 2004
  • To develop the enzyme-linked immunosorbent assay (ELISA) for the analysis of N-acetylehitooligosaccharides (NACOS) and chitooligosaccharides (COS), specific antibodies (Abs) were produced, and their properties were compared. N-acetylehitohexaose (NACOS6), chitohexaose (COS6), and COS mixture (COSM) conjugated to bovine serum albumin (BSA) were used to immunize rabbits. By the use of specific Abs and NACOS6-horseradish peroxidase (HRP), COS6-HRP, and COSM-HRP conjugates, competitive direct ELISA (cdELISA) was developed. The detection limits of NACOS6 by the anti-NACOS6 Ab and COS6 by the anti-COS6 and the anti-COSM Abs in the cdELISAs were about 0.2, 2, ana 2 ng/ml (ppb), respectively. In the cdELISA, the anti-NACOS6 Ab was found to recognize NACOS3-NACOS6, but not N-acetyl-D-glucosamine (GlcNAc), NACOS2, and COSs; the anti-COS6 Ab recognized COS2-COS6 and COSM, but not glucosamine (GlcN) and NACOSs. The recognition pattern of the anti-COSM Ab was almost the same as that of the anti- COS6 Ab, except that the former recognized COS2 and COS3 slightly better than the latter.

Design, Synthesis and Preliminary Biological Evaluation of a Biotin-S-S-Phosphine Reagent

  • Kang, Dong W.;Kim, Eun J.
    • Bulletin of the Korean Chemical Society
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    • v.35 no.2
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    • pp.383-391
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    • 2014
  • Biotin-S-S-Phosphine was designed and synthesized as a potential tool for a proteomic study of O-GlcNAcmodified proteins. This reagent features a disulfide linker between a triarylphosphine moiety, which allows selective conjugation to azide-containing proteins, and a biotin moiety that can allow easy isolation through its strong affinity toward avidin-coated solid beads. The disulfide linkage within this reagent can allow the easy release of the bound molecules of interest, which is difficult to achieve when a biotin:avidin pair is used alone, by reducing the disulfide bond of the reagent with DTT. Preliminary in vitro biological assays with azidelabeled and unlabeled cell lysates and a pure protein Nup62 showed that the Biotin-S-S-Phosphine reagent is highly reactive toward the free thiol groups of proteins. When a molecular tool with a disulfide linker is applied to the enrichment of the molecules of interest from other species, it is important to block the free-thiols of the sample using exhaustive alkylation prior to the Staudinger ligation reactions to restore the bioorthogonal nature of this reaction.

Studies on the Residues of Chlorinated Organic Insecticides -3. Heptachlor residues in soil 15 years after yearly treatment of the soil insecticide in a tobacco field- (유기염소계 살충제의 잔류분에 관한 연구 -3. 담배연작토양의 Heptachlor 잔류분에 관하여-)

  • Park, Chang-Kyu
    • Applied Biological Chemistry
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    • v.18 no.2
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    • pp.61-64
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    • 1975
  • Soil residues in a tobacco field yearly treated with a soil insecticide, Heptachlor, was analyzed by GLC equipped with electron capture detector(ECD). In addition, translocation of the heptachlor residues into two staple vegetables, radish and chinese cabbage was also studied under field conditions. The results were summarized as follows; 1. The Heptachlor residues in the soil of tobacco plots were less than 0.010 ppm. 2. The Heptachlor residues in radish and chinese cabbages cultivated in the tobacco plot were also below 0.010 ppm. 3. Amount of Heptachlor residues translocated into the vegetables were not related to the concentrations of the residues in the soil.

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Molecular Simulations and Conformational Studies of Fucoseα1-3)Gal(β1-X)GlcNAc where X=3, 4, or 6 Oligosaccharides

  • Yoo, Eun-Sun;Yoon, In-Mo
    • Bulletin of the Korean Chemical Society
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    • v.29 no.9
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    • pp.1755-1760
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    • 2008
  • Energy minimization and conformational studies of molecular ions generated by ESI (electrospray ionization) tandem mass spectrometry (MS/MS) can be used for the discrimination of stereoisomeric permethylated and sodium cationized trisaccharides. Sets of fucose-containing trisaccharides having different internal and terminal linkages have been synthesized to analyze the reducing terminal linkage positions using BT and IT fusion approaches. A detailed investigation has been undertaken on the conformational behaviors of four trisaccharide fragments from human milk and blood group determinants of Type 1 and Type 2, namely Fuc($\alpha$1- 3)Gal($\beta$1-3)GalNAc and Fuc($\alpha$1-3)Gal($\beta$1-X)GlcNAc where X = 3, 4 and 6 using molecular modeling methods. Three dimensional rigid and adiabatic phi-psi-energy maps (Surfer program) describing the energy as a function of rotation around corresponding glycosidic linkages were calculated by SYBYL molecular modeling and MM4 force field programs conjunction with cleavage energies of ESI MS/MS for the side group orientations. This approach predicted conformational behaviors exhibited by isomer saccharides for future applications on biologically active glycoconjugates and to exploit a faster method of synthesizing a series of structural isomeric oligosaccharides.

Biochemical Characterization of a Glycosyltransferase Homolog from an Oral Pathogen Fusobacterium nucleatum as a Human Glycan-Modifying Enzyme

  • Kim, Seong-Hun;Oh, Doo-Byoung;Kwon, Oh-Suk;Jung, Jae-Kap;Lee, Yun-Mi;Ko, Ki-Sung;Ko, Jeong-Heon;Kang, Hyun-Ah
    • Journal of Microbiology and Biotechnology
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    • v.18 no.5
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    • pp.859-865
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    • 2008
  • Bacterial glycosyltransferases have drawn growing attention as economical enzymes for oligosaccharide synthesis, with their easy expression and relatively broad substrate specificity. Here, we characterized a glycosyltransferase homolog (Fnu_GT) from a human oral pathogen, Fusobacterium nucleatum. Bioinformatic analysis showed that Fnu_GT belongs to the glycosyltransferases family II. The recombinant Fnu_GT (rFnu_GT) expressed in Escherichia coli displayed the highest glycosylation activity when UDP-galactose (Gal) was used as a donor nucleotide-sugar with heptose or N-acetylglucosamine (GlcNAc) as an acceptor sugar. Interestingly, rFnu_GT transferred the galactose moiety of UDP-Gal to a nonreducing terminal GlcNAc attached to the trimannosyl core glycan, indicating its potential as an enzyme for human-type N-glycan synthesis.

An Adjustment for a Regional Incongruity in Global land Cover Map: case of Korea

  • Park Youn-Young;Han Kyung-Soo;Yeom Jong-Min;Suh Yong-Cheol
    • Korean Journal of Remote Sensing
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    • v.22 no.3
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    • pp.199-209
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    • 2006
  • The Global Land Cover 2000 (GLC 200) project, as a most recent issue, is to provide for the year 2000 a harmonized land cover database over the whole globe. The classifications were performed according to continental or regional scales by corresponding organization using the data of VEGETATION sensor onboard the SPOT4 Satellite. Even if the global land cover classification for Asia provided by Chiba University showed a good accuracy in whole Asian area, some problems were detected in Korean region. Therefore, the construction of new land cover database over Korea is strongly required using more recent data set. The present study focuses on the development of a new upgraded land cover map at 1 km resolution over Korea considering the widely used K-means clustering, which is one of unsupervised classification technique using distance function for land surface pattern classification, and the principal components transformation. It is based on data sets from the Earth observing system SPOT4/VEGETATION. Newly classified land cover was compared with GLC 2000 for Korean peninsula to access how well classification performed using confusion matrix.

Triglyceride Composition of Some Vegetable Oils 1. Triglyceride Composition of Sesame Oil (식물유의 Triglyceride 조성 1. 참기름의 Triglyceride 조성)

  • Park Yeung-Ho;WADA Shun;KOIZUMI Chiaki
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.14 no.1
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    • pp.1-6
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    • 1981
  • In this study, sesame oil was chosen as the experimental sample and analysed for its triglyceride composition by high-performance liquid chromatography(HPLC) in combination with gas liquid chromatography(GLC). The triglycerides were separated from sesame oil by liquid chromatographies on Bio-Beads SX-2 and on Sephadex LH-20, and fractionated into five groups on the basis of their partition numbers by reverse phase HPLC on a column packed with $\mu-Bondapak$ C18 using methanol-chloroform mix-ture as a solvent. Each of these collected fractions gave one to three peaks in the GLC chromatograms according to the acyl carbon number of the triglyceride, and fatty acid composition of the triglyceride was also analysed by GLC. From the results, it was found that the sesame oil consists with twenty one kinds of triglyceri-des, and the major triglycerides of sesame oil are those of $(2\;{\times}\;C18:1,\;C18:2\;;\;17.1\%),\;(C18:1,\;2{\times}C18:2\;;\;17.0\%),$ $(3\;{\times}\;C18:2\;;\;17.0\%),\;(3\;{\times}\;C18:1\;;\;10.9\%),$ $(3\;{\times}\;C18:2\;;\;9.6\%),\;(C16:0,\;C18:1,C18:2\;;\;7.9\%),$ $(C16:0,\;2\;{\times}\;C18:1\;;\;7.4\%),\;(C16:0,\;2\;{\times}\;C18:2\;;\;6.8\%),$ $(C18:0,\;C18:1,\;C18:2\;;\;3.1\%),\;(2\;{\times}\;C18:0,\;C18:2\;;\;1.5\%)$ $(C18:0,\;2\;{\times}\;C18:1\;;\;1.4\%),\;(C16:0,\;C18:0,\;C18:1\;;\;1.3\%),$ $(2\;{\times}\;C16:0,\;C18:1\;;\;1.2\%),\;and\;(C16:0,C18:0,\;C18:2\;;\;1.0\%)$.

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Characterization of a Mucolipidosis Type II Mouse Model and Therapeutic Implication of Lysosomal Enzyme Enriched Fraction Derived from Placenta (뮤코지방증 2형 마우스 모델의 특징과 태반에서 추출한 리소좀 효소 투여의 결과)

  • Cho, Sung Yoon;Kim, Ki-Yong;Kim, Su Jin;Sohn, Young Bae;Maeng, Se Hyun;Kim, Chi Hwa;Ko, Ah-Ra;Song, Junghan;Yeau, Sung-Hee;Kim, Kyung-Hyo;Jin, Dong-Kyu
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.12 no.1
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    • pp.5-13
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    • 2012
  • I-cell disease (mucolipidosis type II; MIM 252500) and pseudo-Hurler polydystrophy (mucolipidosis type III; MIM 252600) are disorders caused by abnormal lysosomal transport in cells. The presence of numerous inclusion bodies in the cytoplasm of fibroblasts, a lack of mucopolysacchariduria, increased lysosomal enzyme activity in serum, and decreased GlcNAc-phosphotransferase activity are hallmark. Here, we attempted to investigate phenotypical and biochemical characteristics of the knockoutmouse of GlcNAc-phosphotransferase ${\alpha}/{\beta}$ subunits; in addition, we also attempted to determine whether the lysosome enriched fraction derived from placenta can be beneficial to phenotype and biochemistry of the knockout mouse.We found that the knockout mouse failed to thrive and had low bone density, as is the case in human. In addition, skin fibroblasts from the animal had the same biochemical characteristics, including increased lysosomal enzyme activity in the culture media, in contrast to the relatively low enzyme activity within the cells. Intravenous injection of the lysosome rich fraction derived from placenta into the tail vein of the animal resulted in a gain of weight, while saline injected animals didn't.In conclusion, our study demonstrated the phenotypical and biochemical similarities of the knockout mouse to a mucolipidosis type II patient and showed the therapeutic potential of the lysosome enriched fraction. We admit that a larger scale animal study will be needed; however, the disease model and the therapeutic potential of the lysosome enriched fraction will highlight the hope for a novel treatment approach to mucopolipidosis type II, for which no therapeutic modality is available.

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Studies on the Triglyceride Composition of Some Vegetable Oils -IV. Triglyceride Composition of Rice Bran Oil- (식물유(植物油)의 Triglyceride조성(組成)에 관한 연구(硏究) -제(第) 4 보(報) : 쌀겨기름의 Triglyceride조성(組成)-)

  • Choi, Su-An;Park, Yeung-Ho
    • Korean Journal of Food Science and Technology
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    • v.15 no.2
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    • pp.108-111
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    • 1983
  • High performance liquid chromatography (HPLC) was applied to the analysis of triglycerides of rice bran oil. The triglycerides were clearly separated in five peaks by HPLC on a column packed with ${\mu}-Bondapack$ C18 using methanol-chloroform mixture as a solvent. Compositions of the triglyceride and fatty acid of the fraction was also analyzed by gas liquid chromatography (GLC). Each of these collected fractions gave three to four peaks in the GLC chromatograms according to the carbon number of the triglyceride. The fitty acid compositions of these triglycerides were mainly composed of C16:0, C18:1 and C18:2 fatty acids. The major triglycerides of the rice bran oil were found to be those of (C16:0, C18:1, C18:2;16.64%), $(2{\times}C18:1,\;C18:2;16.18%)$, $(3{\times}C18:1;13.7%)$, $(C16:0,\;2{\times}C18:1;12.77%)$,$(C18:1,\;2{\times}C18:2;9.16%)$ and $(C16:0,\;2{\times}C18:2;6.42%)$

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Polymorphism in the intron 20 of porcine O-linked N-acetylglucosamine transferase

  • Kim, Jong Gug;Nonneman, Dan;Kim, Doo-Wan;Shin, Sangsu;Rohrer, Gary A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.8
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    • pp.1086-1092
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    • 2017
  • Objective: O-linked N-acetylglucosamine (O-GlcNAc) transferase (OGT) catalyzes the addition of O-GlcNAc and GlcNAcylation has extensive crosstalk with phosphorylation to regulate signaling and transcription. Pig OGT is located near the region of chromosome X that affects follicle stimulating hormone level and testes size. The objective of this study was to find the variations of OGT between European and Chinese pigs. Methods: Pigs were tested initially for polymorphism in OGT among European and Chinese pigs by polymerase chain reaction and sequencing at the U.S. Meat Animal Research Center (USMARC). The polymorphism was also determined in an independent population of pigs including European and Chinese Meishan (ME) breeds at the National Institute of Animal Science (NIAS, RDA, Korea). Results: The intron 20 of OGT from European and Chinese pigs was 514 and 233 bp, respectively, in the pigs tested initially. They included 1 White composite (WC) boar and 7 sows ($2Minzu{\times}WC$, $2Duroc\;[DU]{\times}WC$, $2ME{\times}WC$, $1Fengzing{\times}WC$) at USMARC. The 281-bp difference was due to an inserted 276-bp element and GACTT in European pigs. When additional WC and ME boars, the grandparents that were used to generate the $1/2ME{\times}1/2WC$ parents, and the 84 boars of 16 litters from mating of $1/2ME{\times}1/2WC$ parents were analyzed, the breeds of origin of X chromosome quantitative trait locus (QTL) were confirmed. The polymorphism was determined in an independent population of pigs including DU, Landrace, Yorkshire, and ME breeds at NIAS. OGT was placed at position 67 cM on the chromosome X of the USMARC swine linkage map. Conclusion: There was complete concordance with the insertion in European pigs at USMARC and NIAS. This polymorphism could be a useful marker to identify the breed of origin of X chromosome QTL in pigs produced by crossbreeding Chinese and European pigs.