• Title/Summary/Keyword: Carbohydrate column

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Extraction of Glycosaminoglycan from Sea Hare, Aplysia kurodai, and Its Functional Properties 2. Structural Properties of Purified Glycosaminoglycan (군소(Aplysia kurodai)에 분포하는 글루코사미노글리칸의 추출과 기능특성 2. 글루코사미노글리칸의 구조 특성)

  • Yoon, Bo-Yeong;Choi, Byeong-Dai;Bae, Dong-Won;Choi, Yeung-Joon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.11
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    • pp.1647-1653
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    • 2010
  • Glycosaminoglycan (GAG) was purified from polysaccharide extracted from sea hare muscle on DEAE-Sepharose column and investigated for the functional groups, distribution of sugars, composition of disaccharide and structure of GAG. Purified GAG was composed of disaccharide above 55% of total sugar. Purified GAG showed amide I peak in 1648/cm and C-O stretch peak as properties of carbohydrate, amino acid peak in 1457/cm, and peak in 866/cm as properties of monosaccharide by FT-IR. Fucose, N-acetylgalactosamine, N-acetylglucosamine, glucose, galactose, mannose and xylose were found in MALDI-TOF MS/MS spectra of hydrolysates by chondroitin sulfate ABC lyase and heparanase I. Purified GAG was expected to be heparan sulfate including N-acetylgalactosamine and N-acetylglucosamine above 70% of total sugar. The structure of GAG was supposed as GlyUA(2S)-GlcNS and GlyUA-GlcNS(6S) with O-linkage on protein core.

Production and Action of Microbial Piscicidal Substance (미생물에 의한 살어성물질의 생성 및 그 작용)

  • 도재호;서정훈
    • Microbiology and Biotechnology Letters
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    • v.6 no.1
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    • pp.41-46
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    • 1978
  • Piscicidal substance produced by Streptomyces sp. isolated from soil was toxic against various kinds of fish. After extraction with CH$Cl_3$ from the culture medium, the substance was purified by avicel column chromatography. In order to test toxicity, various kinds of fish were subjected to the acqueous solution of 100 us of the substance per liter of water. Generally, the substance was toxic to most fish, but Macropodus chinenes and Misgurnus mizolepis are resistant to the substance than Gobius similis and Pseudorasbora parva. The substance was stable at pH range, 3.0 to 7.0, but labile at alkaline pH, and to heat as well. Succinic dehydrogenase on most of tissue cell of Cyprinus carpio was inhibited by this substance strongly, but spinal cord was not inhibited. By addition of Cu and Pb salts to the culture medium, piscicidal substance producibility was activated.

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Inhibitory Activity of Garlic Fermented by Pediococcus pentosaceus KACC 91419 against Antibiotic-resistant Pathogens

  • Ham, Jun-Sang;Lee, Seung-Gyu;Kim, Min-Kyung;Oh, Mi-Hwa;Jeong, Seok-Geun;Kim, Dong-Hun;Lee, Se-Hyung;Chae, Jong-Pyo;Lee, Ji-Yoon;Kang, Dae-Kyung
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.9
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    • pp.1236-1243
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    • 2010
  • The aim of this study was to screen lactic acid bacteria for the fermentation of garlic and to assess the increase in inhibitory activity of garlic fermented against antibiotic-resistant pathogens for use as an animal feed supplement. We screened 45 strains of lactobacillus for the fermentation of garlic. Of these strains, 23 showed similar growth rates with or without allicin. Cultures of the 23 strains were mixed with an equivalent amount of garlic juice and incubated overnight at $37^{\circ}C$. The three strains with the lowest pH values were Lactobacillus paracasei KCTC 3169, L5 strain, and L. reuteri SW. Garlic juice fermented by the L5 strain more strongly inhibited antibiotic-resistant pathogenic bacteria than L. paracasei KCTC 3169, L. reuteri SW, or garlic juice itself. By examining carbohydrate utilization, morphologic properties and 16S rRNA gene sequences, we identified the L5 strain as Pediococcus pentosaceus and deposited it in the name of P. pentosaceus KACC 91419 into the Korea Agricultural Culture Collection. To identify the antimicrobial compound from the garlic filtrate fermented by P. pentosaceus KACC 91419, we fractionated P. pentosaceus KACC 91419 culture on a C18 column and checked the antimicrobial activity of fractions A6 to A10. Only fraction A9 showed inhibitory activity on Staphylococcus aureus. Comparing the mass spectra of the fractions with and without antimicrobial activity, we observed a single dominant product ion (m/z 157.99) from the fraction showing antimicrobial activity. Its molecular mass (157.99) was 2 atomic mass units less than that of allicin (162.02). This suggests that allicin might be converted to its derivative, which has antimicrobial activity, during fermentation by P. pentosaceus KACC 91419.

Cloning and Characterization of Xylanase 11B Gene from Paenibacillus woosongensis (Paenibacillus woosongensis의 Xylanase 11B 유전자 클로닝과 특성분석)

  • Yoon, Ki-Hong
    • Microbiology and Biotechnology Letters
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    • v.45 no.2
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    • pp.155-161
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    • 2017
  • A gene coding for the xylanase predicted from the partial genomic sequence of Paenibacillus woosongensis was cloned by PCR amplification and sequenced completely. This xylanase gene, designated xyn11B, consisted of 1,071 nucleotides encoding a polypeptide of 356 amino acid residues. Based on the deduced amino acid sequence, Xyn11B was identified to be a modular enzyme, including a single carbohydrate-binding module besides the catalytic domain, and was highly homologous to xylanases belonging to glycosyl hydrolase family 11. The SignalP4.1 server predicted a stretch of 26 residues in the N-terminus to be the signal peptide. Using DEAE-Sepharose and Phenyl-Sepharose column chromatography, Xyn11B was partially purified from the cell-free extract of recombinant Escherichia coli carrying a copy of the P. woosongensis xyn11B gene. The partially purified Xyn11B protein showed maximal activity at $50^{\circ}C$ and pH 6.5. The enzyme was more active on arabinoxylan than on oat spelt xylan and birchwood xylan, whereas it did not exhibit activity towards carboxymethylcellulose, mannan, and para-nitrophenyl-${\beta}$-xylopyranoside. The activity of Xyn11B was slightly increased by $Ca^{2+}$ and $Mg^{2+}$, but was significantly inhibited by $Cu^{2+}$, $Ni^{2+}$, $Fe^{3+}$, and $Mn^{2+}$, and completely inhibited by SDS.

Isolation of Mitogenic Glycophosphopeptides from Cheese Whey Protein Concentrate (유청 단백질에서 유도되는 생리활성 펩타이드에 관한 연구)

  • Yun, Sung-Seob
    • Journal of Dairy Science and Biotechnology
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    • v.15 no.1
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    • pp.33-44
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    • 1997
  • We investigated the immunological function of cheese whey protein concentrate (CWPC), which is a by-product of cheese production, using mitogenic activity in murine splenocytes as an index. A fraction isolated by gel filtration and anion exchange chromatography of CWPC showed high mitogenic activity, comparable to the activity of lipopolysaccharide (LPS). The fraction was detected as a single band on SDS-PAGE. It contained calcium, inorganic phosphorus, and carbo-hydrate, indicating the active component to be a glycophosphopeptide (GPP) Since pronase digestion of GPP did not reduce its mitogenic activity, carbohydrate rather than peptide may be important in the activity, When applied on an anti-${\beta}$-caseinophosphopeptide (${\beta}$-CPP ) antibody affinity column, the GPP was separated into two components, one with affinity to ${\beta}$-CPP and the other without such affinity. Both the components contained N-linked oligosaccharide chains and had the mitogenic activity. These results demonstrate that cheese whey contains a GPP having strong mitogenic activity

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High Performance Liquid Chromatographic Determination of Free Sugars in Ginseng and Its Products (고속액체(高速液體) 크로마토그래피에 의(依)한 인삼(人蔘) 및 인삼제품중(人蔘製品中)의 유리당(遊離糖)의 정량(定量))

  • Choi, Jin-Ho;Jang, Jin-Gyu;Park, Kil-Dong;Park, Myung-Han;Oh, Sung-Ki
    • Korean Journal of Food Science and Technology
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    • v.13 no.2
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    • pp.107-113
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    • 1981
  • Free sugars were isolated from ginseng root and its products and analyzed by using high performance liquid chromatogrphy. To isolate free sugars from aqueous sample solution fat-soluble components, crude saponin and protein were removed from the solution by extracting with benzene, water-saturated butanol and 80% ethanol, respectively. Free sugars found from both ginseng root and its products were fructose, glucose, sucrose and maltose, and the only sugar detected from red ginseng root and its products was rhamnose. Major sugar detected from fresh ginseng and white ginseng roots was sucrose, while sucrose and maltose were major sugars of red ginseng root.

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Determination of Isomaltooligosaccharides in Yoghurts by Using HPLC-ELSD (HPLC-ELSD를 이용한 발효유 제품 중의 Isomaltooligosaccharides 분석법 개발)

  • Ko, Jinhyouk;Lee, Moon-Seok;Kwak, Byung-Man;Ahn, Jang-Hyuk;Park, Jong-Su;Kwon, Joong-Ho
    • Food Science of Animal Resources
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    • v.33 no.3
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    • pp.417-424
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    • 2013
  • A rapid and simple analytical method for the determination of 9 isomaltooligosaccharides (IMO) species in yoghurts was developed using dispersive solid phase extraction (dSPE) clean-up technic and high performance liquid chromatography with evaporative light-scattering detector (HPLC-ELSD). In this study, 9 IMO were extracted from samples simply with chemical reagent using ISO22662 IDF198 method and additional dSPE clean-up. The optimum instrument conditions for the determination were used carbohydrate ES $5{\mu}$ column with gradient elution of water and acetonitrile and ELS detector. The linearity of this method was expressed as the correlation coefficient ($r^2$), the results of IMO 9 species were shown in 0.9999. LOD and LOQ were respectively 7.9-22.1 mg/kg, 25.9-72.8 mg/kg. The accuracy of intra- and inter-day measurements were in the range from $84.3{\pm}4.5$ to $104.9{\pm}6.5%$, and the preceision of the intra- and inter-day measurements were in the range from 0.8 to 7.7%. The recoveries were from $84.3{\pm}4.5$ to $104.9{\pm}6.5%$. The determination results of IMO 9 species for the 9 yoghurts circulated in the market were in the range from $0.317{\pm}0.007$ to $1.624{\pm}0.050$ g/100 g. The newly developed method is appropriate for the determination of IMO in yoghurts, is a rapid and simple method with excellent resolution in compared with previous method.

Chemical Composition and Biological Activities of Immunostimulants Purified from Alkali Extract of Poria cocos Sclerotium (복령 균핵의 알칼리추출물에서 정제한 면역활성 증강물질의 작용과 화학구성)

  • Rhee, Sang-Dal;Cho, Soo-Muk;Park, Jeong-Sik;Han, Sang-Bae;Jeon, Young-Jin;Kim, Hwan-Mook;Kim, Gwang-Po
    • The Korean Journal of Mycology
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    • v.27 no.4 s.91
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    • pp.293-298
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    • 1999
  • An one percent sodium carbonate extract prepared from sclerotia of Poria cocos activated the proliferation of the T lymphocytes as measured by mixed lymphocyte responses(MLR). The active fraction, PCSC22, was isolated from an one percent sodium carbonate extract by a combination of fractionation procedures, including ethanol precipitation and chromatographies on column of DEAE-cellulose and Sephadex G50. Carbohydrate and peptide contained in PCSC22 were 78 : 22% in ratio. On employing gel filtration high performance liquid chromatography, PCSC22 exhitited a homogeneous peak with an average molecular weight of 8 kDa. The sugar moiety of PCSC22 was composed with mannose (92%), galactose (6.2%) and arabinose (1.3%), which might be indicated as heteromannan. Fifteen amino acids were found in peptide moiety of the polysaccharide and aspartic acid, serine, and valine were major components. PCSC22 activated the primary proliferation of T lymphocytes measured by mixed lymphocyte responses, the antibody production of the B lymphocytes and the secretion of nitric oxide from macrophage cell line, RAW264.7.

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Purification of ${\alpha}-Amylase$ Inhibitor from White Kidney Bean(Phaseolus vulgaris) (White Kidney bean(Phaseolus vulgaris)로부터 ${\alpha}-Amylase$ 저해제의 분리.정제)

  • Chun, Seong-Ho;Ryu, Il-Hwan;Park, Seung-Taeck;Lee, Kap-Sang
    • Korean Journal of Food Science and Technology
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    • v.33 no.1
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    • pp.117-121
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    • 2001
  • White kidney bean, Phaseolus vulgaris, contains proteinaceous inhibitors of ${\alpha}-amylase$. Two inhibitors have been purified by conventional protein fractionation methods such as ethanol precipitation, ammonium sulfate fractionation, DEAE-Sephadex ion exchange chromatography and Sephadex G-100 gel chromatography. The inhibitors were purified as I-1 and I-2 based on their elution order from the DEAE-Sephadex column. The overall purification ratio were about 15.0 and 14.8 for I-1 and I-2, respectively. The molecular weights of purified ${\alpha}-amylase$ inhibitors were 50,000 and 45,000 determined by Sephadex G-100 gel filtration and SDS-polyacrylamide gel electrophoresis. They contain $17.6{\sim}17%$ of carbohydrate, $70{\sim}80%$ of protein. The carbohydrates were composed of glucose : xylose : mannose : N-acetylglucosamine (5 : 3 : 50 : 42).

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Changes of saccharide content in fruitbody composition of Flammulina velutipes during storage (팽이버섯의 저장기간 및 품종별 자실체내의 당 함량 변화)

  • Jhune, Chang-Sung;Yun, Hyung-Sik;Leem, Hoon-Tae;Kong, Won-Sik;Lee, Kang-Hyo;Sung, Gi-Ho;Cho, Jae-Han
    • Journal of Mushroom
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    • v.9 no.3
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    • pp.123-131
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    • 2011
  • Winter mushroom was monitored to investigate the influence of storage temperature on its quality during the storage and distribution phase. In measuring its quality, the contents of saccharides were quantified with its fruiting bodies using HPLC. Although it has been known to be difficult to separate saccharide isomers, our results indicated that Grace Prevail carbohydrate ES $5{\mu}column$ was the best in the separation to analyze the saccharide out of six columns used in this study. In our results, xylose was the main component of saccharide in the fruiting body of winter mushroom(White line mushroom:47.68mg/g, brown line mushroom: 63.28mg/g). In long-term storage, the total amount of saccharide tended to increase, but trehalose content of the disaccharide decreased. In comparison with the paramount amount of lactose and myo-inositol contents in long-term storage at $4^{\circ}C$, lactose wasn't detected when stored at $-1^{\circ}C$.