• Title/Summary/Keyword: Carbohydrate column

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Immunostimulating Lectins from Marine Natural Products: Characteristics of the MLA-I, MLA-II and MLA-III (해양 천연물로부터 면역기능 조정제 렉틴 개발 : MLA-I, MLA-II, MLA-III의 특성)

  • 정시련;김장환;전경희
    • YAKHAK HOEJI
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    • v.39 no.3
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    • pp.243-251
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    • 1995
  • Three new lectins, MLA-I, MLA-II and MLA-III, have been isolatedand purified from the hemolymph of Meretrix lusoria and reported previously. Biophysicochemical characteristics were investigated with these three MLA lectins. The MLA lectins agglutinated human erythrocytes non specifically and proved as D-galactose group carbohydrate specific. Molecular weight of ML.A-I. II and III were estimated to be 330, 500 and 310KD, respectively, by gel filtration on Sepharose CL-6B column. On SDS-polyacrylamide gel electrophoresis, ML.A-I was dissociated into a single subunit of 42KD, MLA-II was into the twelve subunits of 46, 32, 30, 28, 25, 23. 22, 20. 19, 16, 15, and 14KD, and MLA-III was into the two subunits of 72 and 44KD. The pl of MLA-I, II, III were 4.0. 4.9 and 5.0. Amino acid analysis revealed a high contents of acidic and hydroxy amino acids, and a paucity of sulfur containing amino acids. Proline was not contained in MLA-II.

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Crystal Structure of a Maltogenic Amylase: Insights into a Catalytic Versatility

  • Oh, Sang-Taek;Cha, Sun-Shin;Kim, Hyun-Ju;Kim, Tae-Jip;Cho, Hyun-Soo;Park, Kwan-Hwa;Oh, Byung-Ha
    • Proceedings of the Korean Biophysical Society Conference
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    • 1999.06a
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    • pp.35-35
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    • 1999
  • Amylases catalyze the hydrolysis of starch material and play central roles in carbohydrate metabolism. The structure and a size exclusion column chromatography proved that the enzyme is a dimer in solution. The N -terminal segment of the enzyme folds into a distinct domain and comprises the enzyme active site together with the central (${\alpha}$/ ${\beta}$)$\sub$8/ barrel of the adjacent subunit.(omitted)

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Purification of $\alpha$-Amylase Inhibitor from Naked Barley in Korea (한국산 쌀보리 $\alpha$-Amylase 저해물질의 분리 및 정제)

  • 심기환;문주석;신창식;최진상;박석규
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.4
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    • pp.556-562
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    • 1995
  • The $\alpha$-amylase inhibitor from naked barley was purified by DEAE-cellulose, Concanavalin-A sepharose and superose 6 column chromatography, and confirmed by capillary electrophoresis. The purified $\alpha$-amylase inhibitor showed a single band of 29KD in molecular weight when estimated by the SDS-PAGE. Its purity was increased by 12-fold as compared to its crude extract, and its specific activity was found to be 336.7units/mg. The major amino acids of the $\alpha$-amylase inhibitor from naked barley was appeared to be glutamic acid, asparitic acid and arginine. The inhibitor from naked barley was glycoproteins and carbohydrate content of inhibitor was 1.0%.

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Effect of Feeding Frequency of Extruded Diets Containing Different Macro-nutrient Levels on Apparent Nutrient Digestibility in Grower Flounder Paralichthys olivaceus (영양소 함량이 다른 부상 배합사료의 공급횟수가 육성기 넙치의 영양소 소화율에 미치는 영향)

  • Seo, Joo-Young;Choi, Kyoung-Hyun;Choi, Jin;Lee, Sang-Min
    • Journal of Aquaculture
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    • v.18 no.3
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    • pp.160-166
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    • 2005
  • Two feeding trials were carried out to investigate apparent nutrient digestibility of flounder fed experimental diets containing different levels of macro-nutrients by satiation feeding rate (Exp-1) and feeding frequency (Exp-2). Triplicate groups of fish averaging 280 g were fed three experimental diets which contained different levels of carbohydrate, protein and lipid by two feeding regimes (satiation and 80% satiation) and four feeding frequencies (three meals a day, two meals a day, one meal a day and one meal every two days). Feces were collected using a fecal collection column attached to fish rearing tanks for 6 weeks. Apparent digestibilities of dry matter, protein, lipid, energy and carbohydrate were not affected by feeding satiation rate in Exp-1. Apparent protein digestibility was not affected by feeding frequency, whereas affected by dietary composition in Exp-2. Apparent protein digestibility of fish fed a high-protein diet showed a tendency to become higher compared to that of fish fed high-carbohydrate diet and high-lipid diet at the same feeding frequency. Apparent lipid digestibility was not affected by dietary composition, however, affected by feeding frequency. Apparent digestibilities of energy and carbohydrate were affected by both dietary composition and feeding frequency. Apparent digestibities of energy and carbohydrate in fish fed the high-protein diet showed a tendency to become higher compared to that of fish fed the high-carbohydrate diet and high-lipid diet at the same feeding frequency. Apparent digestibities of energy and carbohydrate tended to decrease with increasing of feeding frequency at the same dietary composition.

Analysis of Sugar Chain Structure of PAS-7 Glycoprotein from Bovine Milk Fat Globule Membrane by US RAAM 2000 (OGS RAAM2000을 이용한 유지방구막 PAS-7 당단백질의 당쇄구조 해석)

  • 석진석
    • Food Science of Animal Resources
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    • v.21 no.4
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    • pp.367-373
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    • 2001
  • Glycoproteins PAS-6(50 kDa) and -7(47 kDa) from the bovine milk fat globule membrane share a common protein core but differ in their carbohydrate moiety. We have analyzed and proposed the structures of the N-linked sugar chains of PAS-7 by Oxford Glyco System(OGS) RAAM2000. The N-linked sugar chains were liberated from PAS-7 by hydrazinolysis and, after modifying the reducing ends with 2-aminobenzamide(2-AB), were separated into one neutral(7N, 55%) and two acidic(7M, mono-, 43%; 7D, di-, 2%) sugar chain groups. 7N was finally separated into 5 chains(a, b, c, d, and e), respectively. The structure of this 2AB-neutral sugar chain was determined by sugar analysis, exoglycosidase digestion with OGS glycosidase Kit and OGS RAAM2000 system. The results show that fraction e was the same of reported 7N1A, the biantennary complex type with a fucose on reducing end and two N-acetyllactosamine branch on non-reducing end. Therefore, it was proved that OGS RAAM2000 method is in conformity with conventional analysis of sugar chain structure from bovine PAS-7.

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Studies on analysis of the Korean lager beers. (I) "On the estimation of total carbohydrates, amino acids and peptides" (한국산 맥주성분에 관한 연구 (제 1 ) "Total carbohydrates, amino acids 및 peptides 의 정 에 관하여")

  • 한홍의;김종협
    • Korean Journal of Microbiology
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    • v.6 no.2
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    • pp.63-67
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    • 1968
  • Total carbohydrates, amino acids and peptide-like substances in two kinds of Korean lager beers have been analyzed by the calorimetric method of Dreywood's anthrone reagent and Fowden's ninhydrin reagent. The samples were fractionated with column of ion-exchange resin. The experimental results are as follows; 1. Amounts of non-hydrolyzed carbohydrates in the part of column processed is 1. 82% and 1. 96 % (the value was measured by Bertrand's method). But the amounts of those measured by Dreywood's anthrone method are 5.57% and 4.25%, this values are much more than those of Bertrand's method. 2. It can be estimated the amounts of gum and dextrin are 3.75% and 3.30% in both two beers, by comparison of samples with the above mentioned two method. 3. The amounts of carbohydrates by anthrone reagent in acid-hydrolyzed beers are much increased than those of non-hydrolyzed, so it is suggested the presence of polymer carbohydrates which couldn't be detected by Bertrand's method. 4. Total amounts of amino acids are 0.015% and 0.025% (as glycine) in non-hydrolyzed beers measured by ninhydrine color reaction method, on the other hand the amount of amino acids in acid hydrolyzed beers are 0.06% and 0.056%, this is much more than those of nonhydrolysis. The different amounts means that of peptide-like substances. 5. It is necessary to determine the constituent of amino acid for the better taste of beer, and also it is desirable to check the role of carbohydrates in the course of fermentation, mashing and on lager beer for effective utilization of carbohydrate materials to eliminate the losses.

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Possible Mechanism Underlying the Antiherpetic Activity of a Proteoglycan Isolated from the Mycelia of Ganoderma lucidum in Vitro

  • Li, Zubing;Liu, Jing;Zhao, Yifang
    • BMB Reports
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    • v.38 no.1
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    • pp.34-40
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    • 2005
  • GLPG (Ganoderma lucidum proteoglycan) was a bioactive fraction obtained by the liquid fermentation of the mycelia of Ganoderma lucidum, EtOH precipitation, and DEAE-cellulose column chromatography. GLPG was a proteoglycan with a carbohydrate: protein ratio of 10.4: 1. Its antiviral activities against herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) were investigated using a cytopathic inhibition assay. GLPG inhibited cell death in a dose-dependent manner in HSV-infected cells. In addition, it had no cytotoxic effect even at 2 mg/ml. In order to study the mode of action of the antiviral activity of GLPG, cells were treated with GLPG before, during, and after infection, and viral titer in the supernatant of cell culture 48 h post-infection was determined using a $TCID_{50}$ assay. The antiviral effects of GLPG were more remarkable before viral treatment than after treatment. Although the precise mechanism has yet to be defined, our work suggests that GLPG inhibits viral replication by interfering with the early events of viral adsorption and entry into target cells. Thus, this proteoglycan appears to be a candidate anti-HSV agent.

Characterization of a Substance from Photobacterium damsela subsp. piscicida that Non-specifically Binds to Streptavidin

  • Jung Tae Sung;Kim D. Thompson;Adams Aelexandra;Oh Myung Joo
    • Fisheries and Aquatic Sciences
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    • v.3 no.1
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    • pp.52-63
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    • 2000
  • Non-specific reaction has been a problem in doing, especially, research and diagnosis for infectious agents. Avidin-biotin-peroxidase complex (ABC) techniques has widely been used to amplify a reaction. Photobacterium damse1a subsp. piscicdia (formerly Pasteurella piscicida) exhibited a capacity to bind with streptavidin non-specifically. The band, estimated 26 K Da in Western blotted paper, was blocked with biotin but incompletely. In an attempt to explore an involvement of the non-specific substance in attaching piscine cells, cell attachment test performed using anti- Ph. d. subsp piscicida sera raised mouse and rabbit exhibited slightly blocking effects for Mediterranean (1736) and significantly for Japanese (Sp 92144) isolate. Biotin decreased the attachment ability significantly for Sp92144 but it was not effective to 1736. Both isolates showed greatly enhanced attachment ability with poly-L-lysin. The non-specific binding substance was contained in bacterial extracellular products (ECPs). The substance was able to purified with 2-imminobiotin affinity column, the purified substance appeared to have 4 bands in silver staining, and had a carbohydrate branch. This purified substance showed cytotoxic effects selectively between 5 piscine cell lines. Moreover, it stimulated rainbow trout macrophage in terms of reduction of cytochrome cas well as yeast phagocytosis, significantly.

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Isolation and Characterization of Two Korean Mistletoe Lectins

  • Kang, Tae-Bong;Song, Seong-Kyu;Yoon, Taek-Joon;Yoo, Yung-Choon;Lee, Kwan-Hee;Her, Erk;Kim, Jong-Bae
    • BMB Reports
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    • v.40 no.6
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    • pp.959-965
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    • 2007
  • Two isolectins (KML-IIU and the KML-IIL) were individually isolated from the previously reported Korean mistletoe lectin, KML-C, by using an immunoaffinity column. Molecular weights of the KML-IIU and the KML-IIL were 64 kDa and 60 kDa respectively. Both of the lectins were composed of heterogeneous A and B subunits linked with a disulfide bond, and showed the same carbohydrate-binding specificities for Gal and GalNAc. However, they are different not only in biophysical properties (glycosylation and amino acid compositions) but also bioactivities (cell killing and cytokine induction). The KML-IIL showed 17-145 times stronger in cytotoxicities to various human and mouse cancer cell lines than the KML-IIU. The KML-IIL also induced TNF-$\alpha$ secretion from mouse peritoneal macrophages 4.5 times better than the KML-IIU. The results demonstrated isolectins in Korean mistletoe were varied in bioactivities and the KML-IIL may be developed as an anti-cancer agent.

Enzymatic N-glycan analysis of 31 kDa molecule in plerocercoid of Spirometra mansoni (sparganum) and its antigenicity after chemical oxidation

  • Chung, Young-Bae;Kong, Yoon;Yang, Hyun-Jong
    • Parasites, Hosts and Diseases
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    • v.42 no.2
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    • pp.57-60
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    • 2004
  • A highly specific antigenic protein of 31 kDa from plerocercoid of Spirometra mansoni (sparganum) was obtained by gelatin affinity and Mono Q anion-exchange column chromatography. The purified 31 kDa protein was subjected to N-glycan enzymatic digestion for structural analysis. The relative electrophoretic mobility was analyzed by SDS-PAGE, before and after digestion. On SDS-PAGE after enzymatic digestion, the 31 kDa protein showed a molecular shift of approximately 2 kDa, which indicated the possession of complex N-linked oligosaccharides (N-glycosidase F sensitive) but not of high-mannose oligosaccharides (endo-beta-N-acetylglucosaminidase H, non-sensitive). Chemically periodated 31 kDa protein showed statistically non-significant changes with human sparganosis sera by enzyme linked immunosorbent assay (ELISA). Therefore, the dominant epitopes of the 31 kDa molecule in human sparganosis were found to be mainly polypeptide, while N-glycans of the antigenic molecule in sparganum was minimal in anti-carbohydrate antibody production.