• Title/Summary/Keyword: ${\alpha}$-1,3-glucan

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Isolation of $\alpha$-1,3 Glucanase from Microorganism and the Prodution of High Activity $\alpha$-1,3 Glucanase for Hydrolysis of Dental Plaque (치면세균막 분해효소인 $\alpha$-1,3 glucanase를 생산하는 미생물의 분리 및 효소 특성)

  • 조효상;허태련;윤정원
    • Microbiology and Biotechnology Letters
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    • v.21 no.3
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    • pp.263-268
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    • 1993
  • Seventeen strains were isolated from soil, cattle rumen, cereal sewage dregs, insect on agar plate containing insoluble glucan as a sole carbon source from immobilized Streptococcus mutans, which produced alpha-1,3 glucanase for lysis of dental plaque. Among these strains isolated from soil, SW-522 and SW-713 that had appeared to produce the high level of alpha-1,3 glucanase, degraded insoluble glucan from S. mutans 97.6% and 49.4%, respectively in 5 hours. The activity of crude alpha-1,3 glucanase from SW-522 was 1.3mg insoluble glucan/min.mg protein. This enzyme was entirely degraded insoluble glucan on glass tube which produced by S. mutans in TH medium with 5% sucrose.

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(1-3, 1-4)-$\beta$-Glucan and Starch Contents and Their Hydrolytic Enzyme Activities in Developing Barley Kernels (등숙 중인 보리 종실중 (1-3, 1-4)-$\beta$-Glucan과 전분 함량 및 이들의 가수분해효소 활성)

  • 윤성중;박상래;유남희
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.42 no.4
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    • pp.403-409
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    • 1997
  • To obtain information on the accumulation of (1-3, 1-4)-$\beta$-glucans during kernel maturation, (1-3, 1-4)-$\beta$-glucan contents and (1-3, 1-4)-$\beta$-glucanase activities were determined in developing kernels of the two Korean cooking barley varieties, Neulssalbori and Saessalbori. (1-3, 1-4)-$\beta$-Glucan contents in kernels at 5 and 10 days after anthesis(DAA) were very low and the contents increased rapidly in kernels at 15 to 25 DAA. (1-3, 1-4)-$\beta$-Glucan content in kernels at harvest was about 3.5 to 4% of kernel dry matter. (1-3, 1-4)-$\beta$-Glucanase activities were relatively higher in younger kernels but the levels of the activity were very low compared with those in germinating kernels. A significant negative correlation was observed between (1-3, 1-4)-$\beta$-glucan contents and (1-3, 1-4)-$\beta$-glucanase activities. Low levels of (1-3, 1-4)-$\beta$-glucanase activites in kernels at 15 to 30 DAA, however, may indicate that (1-3, 1-4)-$\beta$-glucanases have little effect on the final content of (1-3, 1-4)-$\beta$-glucans in barley kernels. Starch contents and $\alpha$-amylase activities were also determined in developing barley kernels. Starch contents increased rapidly as kernels matured and the content at harvest was about 60% of kernel dry matter. Relativley higher levels of $\alpha$-amylase activities in kernels at the earlier developmental stage decreased rapidly as kernels matured.

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Induction of Dectin-1 Expression and Intracellular Signal Transduction by β-Glucan of Ganoderma lucidum (불로초의 β-Glucan에 의한 Dectin-1 발현 유도와 세포 내 신호전달)

  • Ryu, Han Wook;Kim, Ha Won
    • The Korean Journal of Mycology
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    • v.46 no.2
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    • pp.161-176
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    • 2018
  • Fungal ${\beta}$-glucan, known to have immunostimulatory and antitumor activities, can be recognized by host immune cells as one of the pathogen-associated molecular patterns (PAMPs). Although there are several reports on the diverse immunostimulatory activities of ${\beta}$-glucan, little is known about the intracellular signal transduction of ${\beta}$-glucan. Stimulation of RAW264.7 macrophage cells with ${\beta}$-glucan from Ganoderma lucidum induced the expressions of dectin-1, toll-like receptor 2 (TLR2), TLR4, and TLR6 at the transcription stage. Treatment with ${\beta}$-glucan also induced inflammatory mediators such as macrophage inflammatory proteins (MIP)-$1{\alpha}$, MIP-$1{\beta}$, MIP-$1{\gamma}$, interleukin (IL)-$1{\beta}$, and tumor necrosis factor (TNF)-${\alpha}$. Treatment of the cells with polymyxin B, an inhibitor of lipopolysaccharides (LPS), blocked the induction of inflammatory mediators in LPS- or ${\beta}$-glucan-stimulated systems. Pretreatment of the cells in our cell culture system with LY294002, a phosphoinositide 3-kinase (PI3K) inhibitor, or U0126, a mitogen-activated protein kinase/extracellular-signal-regulated kinase (MAPK/ERK) kinase (MEK)1/MEK2 inhibitor, led to a reduction in the induction of inflammatory mediators in a concentration-dependent manner. These results show that stimulation of the macrophage cells by ${\beta}$-glucan induced the expressions of both dectin-1 and TLRs. We also found that the PI3K/Akt and MEK pathways were involved in the induction of inflammatory mediators in macrophage cells during intracellular signal transduction of ${\beta}$-glucan.

Immunomodulating Activity of Fungal $\beta$-Glucan through Dectin-1 and Toll-like Receptor on Murine Macrophage

  • Kim, Ha-Won
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2006.11a
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    • pp.103-115
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    • 2006
  • $\beta$-Glucan is a glucose polymer that has linkage of $\beta$-(1,3), -(1,4) and -(1,6). As exclusively found in fungal and bacterial cell wall, not in animal, $\beta$-glucans are recognized by innate immune system. Dendritic cells (DC) or macrophages possesses pattern recognition molecule (PRM) for binding $\beta$-glucan as pathogen-associated molecular pattern (PAMP). Recently $\beta$-glucan receptor was cloned from DC and named as dectin-l which belongs to type II C-type lectin family. Human dectin-1 is consisted of 7 exons and 6 introns. The polypeptide of dectin-1 has 247 amino acids and has cytoplasmic, transmembrane, stalk and carbohydrate recognition domains. Dectin-1 could recognize variety of beta-1,3 and/or beta-1,6 glucan linkages, but not alpha-glucans. In our macrophage cell line culture system, dectin-1 mRNA was detected in RA W264.7 cells by reverse transcription-polymerase chain reaction (RT-PCR). Dectin-1 was also detected in the murine organs of spleen, thymus, lung and intestines. Treatment of RA W264.7 cells with $\beta$-glucans of Ganoderma lucidum (GLG) resulted in increased expression of IL-6 and TNF-$\alpha$ in the presence of LPS. However, GLG alone did not increase IL-6 nor TNF-$\alpha$. These results suggest that receptor dectin-1 cooperate with CD14 to activate signal transduction that is very critical in immunoresponse.

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Immunomodulating Activity of Fungal ${\beta}-Glucan$ through Dectin-1 and Toll-like Receptor on Murine Macrophage

  • Kim, Ha-Won
    • 한국약용작물학회:학술대회논문집
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    • 2006.11a
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    • pp.103-115
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    • 2006
  • [ ${\beta}-Glucan$ ] is a glucose polymer that has linkage of ${\beta}-(1,3)$, -(1,4) and -(1,6). As exclusively found in fungal and bacterial cell wall, not in animal, ${\beta}-glucans$ are recognized by innate immune system. Dendritic cells (DC) or macrophages possesses pattern recognition molecule (PRM) for binding ${\beta}-glucans$ as pathogen-associated molecular pattern (PAMP). Recently ${\beta}-glucans$ receptor was cloned from DC and named as dectin-l which belongs to type II C-type lectin family. Human dectin-l is consisted of 7 exons and 6 introns. The polypeptide of dectin-l has 247 amino acids and has cytoplasmic, transmembrane, stalk and carbohydrate recognition domains. Dectin-l could recognize variety of beta-l,3 and/or beta-l,6 glucan linkages, but not alpha-glucans. In our macrophage cell line culture system, dectin-l mRNA was detected in RA W264.7 cells by reverse transcription-polymerase chain reaction (RT-PCR). Dectin-l was also detected in the murine organs of spleen, thymus, lung and intestines. Treatment of RA W264.7 cells with ${\beta}-glucans$ of Ganoderma lucidum (GLG) resulted in increased expression of IL-6 and $TNF-{\alpha}$ in the presence of LPS. However, GLG alone did not increase IL-6 nor $TNF-{\alpha}$ These results suggest that receptor dectin-l cooperate with CD14 to activate signal transduction that is very critical in immunoresponse.

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A Study on the Thermal Characterization of Barley ${\beta}-Glucan$ [mixed-linked $(1-3),(1-4)-{\beta}-D-Glucan$] by Differential Scanning Calorimetry (DSC에 의한 보리 ${\beta}-Glucan$ [mixed-linked$(1-3),(1-4)-{\beta}-D-Glucan$의 열적 특성에 관한 연구)

  • Cha, Hee-Sook;Kim, Mi-Ok;Koo, Sung-Ja
    • Korean Journal of Food Science and Technology
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    • v.25 no.1
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    • pp.22-27
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    • 1993
  • Crude ${\beta}-glucan$ extracted from Barley was purified by stepwide enzyme treatment with thermostable ${\alpha}-amylase$, amyloglucosidase and protease. The thermal properties of Barley ${\beta}-glucan$ were investigated by Differential Scanning Calorimetry. Three endotherms have been observed on DSC thermograms of Barley ${\beta}-glucan$. The first endotherm which produced the gelatinization phenomena commonly observed in Barley ${\beta}-glucan$ became the focus of this study. The temperature range and the enthalpy of gelation exhibited maximum values with increasing concentration of Barley ${\beta}-glucan$. Gelating Barley ${\beta}-glucan$ registered an enthalpy of approximately 0.23 cal/g and exhibited onset temperature (To), peak temperature (Tp) and conclusion temperature (Tc) of $48.8^{\circ}C,\;61.2^{\circ}C\;and\;78.5^{\circ}C$ respectively. The temperature and enthalpy of gelatinizing Barley ${\beta}-glucan$ at both alkali and acid conditions were lower than those at pH 7. With salt present, the Tp and Tc of gelating Barley ${\beta}-glucan$ produced lower temperatures than in conditions where salt was absent, and the enthalpy abruptly decreased. However, increasing salt concentrations did not affect the gelation temperature and the enthalpy of Barley ${\beta}-glucan$. The 'true melting' temperature of Barley ${\beta}-glucan$ was near $184^{\circ}C$ and the melting enthalpy was approximately 34.6 cal/g. The Barley ${\beta}-glucan$ decomposition temperature was in the range of $316^{\circ}C{\sim}346^{\circ}C$.

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Immune-Enhancing Alkali-Soluble Glucans Produced by Wild-Type and Mutant Saccharomyces cerevisiae

  • Ha Chang-Hoon;Lim Ki-Hong;Jang Se-Hwan;Yun Cheol-Won;Paik Hyun-Dong;Kim Seung-Wook;Kang Chang-Won;Chang Hyo-Ihl
    • Journal of Microbiology and Biotechnology
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    • v.16 no.4
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    • pp.576-583
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    • 2006
  • The alkali-soluble glucan of the yeast cell wall contains $\beta-(1,3)-$ and (1,6)-D-linkages and is known to systemically enhance the immune system. In the previous study [6], in order to isolate cell wall mutants, a wild-type strain was mutagenized by exposure to ultraviolet light, and the mutants were then selected via treatment with laminarinase $(endo-\beta-(1,3)-D-glucanase)$. The mass of alkali- and water-soluble glucans produced by the mutant was measured to be 33.8 mg/g of the dry mass of the yeast cell. Our results showed that the mutants generated the amount of alkali-soluble glucan 10-fold higher than that generated by the wild-type. Structural analysis showed that the alkali-soluble glucan from the mutants was associated with a higher degree of $\beta-(1,6)-D-linkage$ than was observed in conjunction with the wild-type. Yeast cell wall $\beta-glucan$ was shown to interact with macrophages via receptors, thereby inducing the release of tumor necrosis factor alpha $(TNF-\alpha)$ and nitric oxide. Alkali-soluble $\beta-glucans$, both from water-soluble and water-insoluble glucan, exhibited a higher degree of macrophage activity with regard to both the secretion of tumor necrosis factor alpha $(TNF-\alpha)$ and nitric oxide and direct phagocytosis, than did the positive control ($1{\mu}g$ of lipopolysaccharide).

Purification of Oat ${\beta}-Glucan$ by ${\alpha}-Amlyase$ Treatment and Characterization of Its Physicochemical Properties (귀리 ${\beta}-glucan$${\alpha}-amlyase$를 이용한 정제와 이화학적 특성)

  • Park, Hee-Joeng;Kang, Tae-Su;Lee, Hee-Bong;Kim, Kwang-Yup;Jang, Keum-Il;Noh, Young-Hee;Jeong, Heon-Sang
    • Korean Journal of Food Science and Technology
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    • v.37 no.5
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    • pp.776-782
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    • 2005
  • The effects of purification using ${\alpha}-amlyase$ (Termamyl 120L) on physicochemical properties of ${\beta}-glucan$ from oat bran were studied. Four fractions were selected as fraction A ($55^{\circ}C$, 15%, pH 6), fraction B ($45^{\circ}C$, 15%, pH 6), fraction C ($50^{\circ}C$, 0%, pH 7), and fraction D ($50^{\circ}C$, 10%, pH 5) from the result of physiological test, and three consecutive subfractions were obtained by repeated ${\alpha}-amlyase$ treatments on the each fractions. The contents of ${\beta}-glucan$, protein, and ash after purification were in 81.4-88.2%, 4.1-6.3% and 2.6-6.2%, respectively. The apparent viscosities of purified ${\beta}-glucan$ aqueous solutions were similar to those of hydroxy methyl cellulose. Glucose was a major monosaccharide of ${\beta}-glucan$ extracts, and xylose and arabinose were also detected as minor constituents on TLC. The average molecular weight ranged $2.0{\times}10^6-5.1{\times}10^6$ and was decreased after purification. From the result of the differential scanning calorimetry, the melting point ranged $130-140^{\circ}C$ with purification step and thermal transition enthalpy was increased. The ratio of ${\beta}-(1{\rightarrow}3)\;to\;{\beta}-(1{\rightarrow}4) $ linkages were 1:2.22-1:2.52, and increased up to 1:5.50 after purification.

Analysis of Immunomodulating Gene Expression by cDNA Microarray in $\beta$-Glucan-treated Murine Macrophage

  • Sung, Su-Kyong;Kim, Ha-Won
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2003.11a
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    • pp.98-98
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    • 2003
  • ${\beta}$-(1,3)-D-Glucans have been known to exhibit antitumor and antimicrobial activities. The presence of dectin-1,${\alpha}$, ${\beta}$-glucan receptor of dendritic cell, on macrophage has been controvertial. RT-PCR analysis led to the detection of dectin-1${\alpha}$ and ${\beta}$ in murine macrophage Raw264.7 cell line. Among the various organs of mouse, dectin-1${\alpha}$ and ${\beta}$ were detected in the thymus, lung, spleen, stomach and intestine. To analyze gene expression modulated by ${\beta}$-glucan treated murine Raw264.7 macrophage, total mRNA was applied to cDNA microarray to interrogate the expression of 7,000 known genes. cDNA chip analysis showed that ${\beta}$-glucan of P. osteatus increased gene expressions of immunomodulating genes, membrane antigenic proteins, chemokine ligands, complements, cytokines, various kinases, lectin associated genes and oncogenes in Raw 264.7 cell line. When treated with ${\beta}$-glucan of P. osteatus and LPS, induction of gene expression of TNF-${\alpha}$ and IFN-R1 was confirmed by RT-PCR analysis. Induction of TNF-R type II expression was confirmed by FACS analysis. IL-6 expression was abolished by EDTA in ${\beta}$-glucan and LPS treated Raw264.7 cell line, indicating that ${\beta}$-glucan binds to dectin-l in a Ca$\^$++/ -dependent manner. To increase antitumor efficacy of ${\beta}$-glucan, ginsenoside Rh2 (GRh2) was co-treated with ${\beta}$-glucan in vivo and in vitro tests. IC$\sub$50/ values of GRh2 were 20 and 25 $\mu\textrm{g}$/$m\ell$ in SNU-1 and B16 melanoma F10 cell line, respectively. Co-treatment with ${\beta}$-glucan and GRh2 showed synergistic antitumor activity with cisplatin and mitomycin C both in vitro and in vivo. Single or co-treatment with ${\beta}$-glucan and GRh2 increased tumor bearing mouse life span. Co-treatment with ${\beta}$-glucan and GRh2 showed more increased life span with mitomycin C than that with cisplatin. Antitumor activities were 67% and 72 % by co-injection with ${\beta}$-glucan and GRh2 in the absence or presence of mitomycin C, respectively.

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Characteristics of Water-soluble Polysaccharide, Showing Inhibiting Activity on ${\alpha}-Glucosidase$, in Cordyceps militaris

  • Chung, Ha-Yull;Yoo, Mi-Kyong;Kawagishi, Hirokazu
    • Food Science and Biotechnology
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    • v.18 no.3
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    • pp.667-671
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    • 2009
  • Extract of water-soluble polysaccharide (CFWx), showing inhibiting activity on ${\alpha}-glucosidase$, was prepared from the fruiting bodies of Cordyceps militaris by hot-water extraction, and ethanol precipitation. Chemical characteristics of CFWx were as follows: carbohydrate content 30% including 16% of uronic acid; 51% protein content; monosaccharide composition, Man:Glu:Gal (30:43:27); molecular weight $3-5{\times}10^4$. CFWx was further purified by ion-exchange, gel-permeation, and affinity chromatography and $CFWx-AH-{\alpha}$ fraction was isolated. Fundamental structure of $CFWx-AH-{\alpha}$ was deduced as ${\alpha}-(1{\to}4$)-D-glucan with ${\alpha}-(1{\to}3$)- and/or ${\alpha}-(1{\to}6$)-D-glycosidic side chains based on methylation analysis.