• Title/Summary/Keyword: Exo-polysaccharide

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Structural elucidation of immuno-stimulating polysaccharide, galactomannan isolated from Colocasia esculenta (토란으로부터 분리한 면역활성 다당 galactomannan의 구조적 특성 규명)

  • Lee, Hee-Won;Shin, Kwang-Soon
    • Korean Journal of Food Science and Technology
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    • v.52 no.6
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    • pp.595-603
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    • 2020
  • To elucidate the structure-function relationship of polysaccharides obtained from Colocasia esculenta, the immuno-stimulating polysaccharide, CE-4a was purified to homogeneity from the crude polysaccharide (CE) extracted from the corms of C. esculenta by two subsequent column chromatographies using DEAE-Sepharose FF and Sephadex G-100, and analysis of their immuno-stimulatory activities and structure were conducted. CE-4a showed an increase in anti-complementary activity in a dose-dependent fashion. The molecular mass was estimated to be 182.4 kDa, which mainly consisted of galactose (43.5%) and mannose (18.2%). Methylation analysis indicated that CE-4a comprised at least 10 different glycosyl linkages, such as terminal Galp, 3-linked Galp, and 4-linked Manp, as well as a characteristic linkage, 2,4,6-branched Manp residue. To analyze the fine structure of CE-4a, it was sequentially digested using endo-α-(1→4)-polygalacturonase, exo-α-galactosidase and endo-β-1,4-D-mannanase. These analyses suggested that CE-4a is to be a highly branched galactomannan with a (1→4)-mannan backbone and galactopyranosyl oligosaccharide side chains.

Structural Analysis of Anti-metastatic Polysaccharides Isolated from Opuntia humifusa (천년초에서 분리한 항전이 다당의 구조 분석)

  • Choi, Jung-Ho;Shin, Kwang-Soon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.2
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    • pp.214-222
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    • 2011
  • To examine the new practical utilization of mucilages in Opuntia humifusa, polysaccharides were isolated from O. humifusa and their anti-metastatic activity and structural analysis were carried out. In experimental lung metastasis of B16BL6 melanoma cells, prophylactically intravenous (i.v.) administration of the crude polysaccharide (CNC-0) from O. humifusa significantly inhibited lung metastasis in a dose-dependant manner. The main polysaccharide, CNC-Ia was purified to homogeneity from CNC-0 by two successive column chromatographies using DEAE-Sepharose FF and Sephadex G-100 and its structure was characterized. Molecular mass of CNC-Ia was estimated to be 700 kDa and it mainly consisted of arabinose, galactose and xylose in addition to two minor sugars such as rhamnose and fucose. Methylation analysis indicated that CNC-Ia comprised at least 18 different glycosyl linkages such as terminal Araf, 5-linked Araf, 4-linked Galp and terminal Xylp in addition to three characteristic linkages such as full branched Araf, 3,4,6-branched Galp and full branched Galp. To analyze the fine structure of CNC-Ia, it was sequentially digested by exo-${\alpha}$-L-arabinofuranosidase and endo-${\beta}$-1,4-D-galactanase. These analyses suggested that CNC-Ia belongs to be a highly branched Type I arabinogalactan which has a ($1{\rightarrow}4$)-${\beta}$-galactan backbone with arabinosyl oligosaccharide side chains.

Intestinal Immune Modulating Polysaccharides of Atractylodes lancea DC. Rhizomes

  • Yu, Kwang-Won
    • Proceedings of the Korean Journal of Food and Nutrition Conference
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    • 2000.05a
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    • pp.1-3
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    • 2000
  • A kind of traditional herbal prescription, Sip-Jeon-Dae-Bo-Tang (TJ-48), has been reported to improve the general condition of cancer patients receiving chemotherapy and /or radiation therapy, and to accelerate hematopoietic recovery from bone marrow injury by mitomycin C. In the present studies, we found that hot-water extract from Atractylodes lancea DC. rhizomes contributed mainly to intestinal immune modulating activity of TJ-48 on Peyer's patch cells mediated-hematopoietic response. After the fractionation, ALR-5 II a-1-1, 5 II b-2-2 and 5 II c-3-1 were further purified from crude polysaccharide fraction. Chemical analyses of each fraction indicated that ALR-5 II a-1-1 mainly contained arabinogalactan fraction whereas ALR-5 II b-2-2 and 5 II c-3-1 mostly comprised pectic polysaccharide fractions as the active polysaccharide ingredients. In order to analyze the essential structure of the activity, ALR-5 II a-1-1 was treated by sequential enzymatic digestion using exo-${\alpha}$-L-arabinofuranosidase and exo-${\beta}$-D-(1\longrightarrow3)-galactanase. Based upon the results of chemical and MALDI-TOF-MS analyses and activity on the digested fractions, the galactosyl side chains consisting of 6-linked Galf and Galp over tetrasaccharide in ALR-5 II a-1-1 might be responsible for the potent intestinal immune modulating activity. To characterize moiety of ALR-5 II c-3-1 for the expression of activity, endo-${\alpha}$-D-(1\longrightarrow4)-polygal acturonase (GL-PGase) purified from dried leaves of Panax ginseng digested ALR-5 II c-3-1. The results of structural analyses and activity on the digested fractions showed that PG-2, which structurally resembles to rhamnogalacturonan II (RG II), and PG-3 (galacturono-oligosaccharides) contained potent intestinal immune modulating activity. Further purification of the other acidic fraction (ALR-5 II b-2-2) indicated that ALR-5 II b-2-2Bb showed that the most potent activity. ALR-5 II b-2-2Bb also contained the unusual component sugars characteristics in RG- II as well as PG-2 derived from ALR-5 II c-3-1, but it could not be digested with GL-PGase. The present studies of relationship between structures and intestinal immune modulating activity of the active polysaccharides purified from A. lancea DC. rhizomes suggested that neutral galactosyl chains consisting mainly of (1\longrightarrow6)-linked Galf and Galp, and RG- II -like moiety with unique component sugars, such as 2-Me-Xyl, 2-Me-Fuc, Api, AceA, Kdo and Dha should play an important role in the potent intestinal immune modulating action of A. lancea DC. rhizomes.

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Effects and Batch Kinetics of Agitation and Aeration on Submerged Cultivation of Ganoderma Iucidum (영지의 액체배양에 미치는 통기.교반의 효과와 동력학적 특성)

  • 이학수;정재현;이신영
    • KSBB Journal
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    • v.16 no.3
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    • pp.307-313
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    • 2001
  • The effects of agitaion and aeration on mycelial growth, exo-polysaccharide (EPS) production, and substrate consumption upon the submerged cultivation of G. lucidum were investigated, and the batch kinetics of the EPS fermentation of G. lucidum were interpreted as function of agitation speed and aeration rate. In a 2.6 L jar fermenter system, the optimum agitation speed and aeration rate for EPS production were determined to be 400 rpm and 1.0 vvm, respectively. The maximum production of EPS obtained was 15 g/L. The logistic model for mycelial growth fitted the experimental data better than that determined by the Monod and the two-thirds power models. The Luedeking-Piret equation adequately modelled the kinetic data obtained for product and substrate.

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Effect of Polyacrylic Acid Addition on Wall Growth in Submerged Cultivation of Ganoderma lucidum (영지 액체 배양의 Wall Growth에 미치는 Polyacrylic Acid의 첨가 효과)

  • Lee, Shin-Young;Lee, Hak-Su
    • Journal of Industrial Technology
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    • v.21 no.A
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    • pp.337-341
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    • 2001
  • This study was carried out to screen the effective polymeric additives preventing wall growth during mycelial submerged cultivation of Ganoderma. lucidum. Effects of additives on mycelial growth and exo-polysaccharide (EPS) production in flask culture and jar fermenter system under 3 different pH processes were investigated, and changes of mycelial morphology were also examined. From flask culture of G. lucidum with additives of different concentrations, 0.1%(w/v) polyacrylic acid was effective for EPS production. As the polyacrylic acid of 0.1%(w/v) was added in medium, wall growth of G. lucidum mycelium grown in jar fermenter system could be protected. The addition of 0.1%(w/v) polyacrylic acid to medium was also improved the mycelial growth and EPS production in the later of submerged culture G. lucidum and no changes of mycelial morphology were observed.

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Yogurt Production Using Exo-polysaccharide-producing Leuconostoc and Weissella Isolates from Kimchi (김치유래 exo-polysaccharide 생성능 Leuconostoc 및 Weissella균을 이용한 발효유 제조)

  • Min, Koung-Ah;Chung, Chang-Ho
    • Korean Journal of Food Science and Technology
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    • v.48 no.3
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    • pp.231-240
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    • 2016
  • The purpose of this study was to investigate the effect of exopolysaccharide (EPS)-producing Leuconostoc and Weissella isolates from kimchi as a probiotic starter and replacement for thickening agents such as pectin and gums in yogurt. Potential probiotic isolates were first screened for their acid and bile tolerance, and then evaluated for antimicrobial activity against Escherichia coli and Salmonella Typhimurium. When the selected Leuconostoc or Weissella isolates were co-inoculated in yogurt without a thickening agent, the yogurt with 4% sucrose produced lower syneresis values than the control and had higher EPS yields. The isolates were able to survive at a level of $10^6CFU/mL$ when incubated at $4^{\circ}C$ for 12 days. This study shows that EPS-producing Leuconostoc and Weissella strains have the potential to produce a synbiotic yogurt.

Cloning and Sequencing of a Gene Involved in the Biosynthesis of Exopolysaccharide in Zoogloea Ramigera 115SLR (Zoogloea Ramigera 115SLR로부터 다당류 생합성에 관여하는 유전자의 분리 및 염기서열 결정)

  • Sam-Pin Lee;Min Yoo
    • Biomedical Science Letters
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    • v.6 no.1
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    • pp.1-9
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    • 2000
  • To identity the genes responsible for the biosynthesis of exopolysaccharide, recombinant plasmids pUEX10 and pLEX10 were constructed from plasmid pLEX3 which was isolated from the recombinant cosmid library of Zoogloea ramigera 115. The complete nucleotide sequence of the 1.7 kb genomic DNA insert in plasmid pUEX10 was determined. Its analysis identified two open reading frames (ORF3 & ORF4) which could encode two proteins. The amino acid sequence derived from ORF3 showed the homology with gumC protein in Xanthomonas campestris as well as exoP protein in Rhizobium melizoti. The partial amino acid sequence of ORF4 showed the homology with polysaccharide export protein in Thermotoga maritima. Z. ramigera 115SLR and Z. ramigera 115SLR/pLEX10 showed the similar pattern for EPS production. Yield of exopolysaccharides produced by Z. ramigera 115SLR and Z. ramigera 115SLR/pLEX10 was 0.26% (w/v) and 0.16% (w/v), respectively.

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Roles of sugar chains in immunostimulatory activity of the polysaccharide isolated from Angelica gigas (참당귀에서 분리한 다당의 면역활성에 대한 당쇄의 역할)

  • Shin, Kwang-Soon
    • Korean Journal of Food Science and Technology
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    • v.51 no.4
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    • pp.336-342
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    • 2019
  • To elucidate structure-function relationship of polysaccharide from Angelica gigas, the AGE-2c-I was purified by two successive chromatography steps. AGE-2c-I showed a potent anti-complementary activity in a dose-dependent manner. AGE-2c-I with a molecular weight of 140 kDa comprised four monosaccharides and 13 glycosyl linkages, and strongly reacted with ${\beta}$-glucosyl Yariv reagent. For the fine structure analysis of AGE-2c-I, it was sequentially digested by exo-arabinofuranosidase and endo-galactanase. The results indicated that AGE-2c-I was a typical RG-I polysaccharide with side chains such as highly branched ${\alpha}$-arabinan, ${\beta}$-($1{\rightarrow}4$)-galactan and arabino-${\beta}$-3,6-galactan. To characterize the active moiety of AGE-2c-I, the anti-complementary activities of AGE-2c-I and its subfractions were assayed. It was observed that the anti-complementary activity of AGE-2c-I was due to the entire structure that resembled RG-I. In addition, arabino-${\beta}$-3,6-galactan side chain (GN-I) in AGE-2c-I probably plays a crucial role in the anti-complementary activity, whereas ${\alpha}$-arabinan side chain (AFN-I) consisting of 5-linked Araf and 3,5-branched Araf partially contributes to the activity.

Effect of Agitation, Aeration and Scale-up on Mycelial Morphology During Liquid Culture of Ganoderma lucidum (영지의 액체배양 중 균사형태에 미치는 통기.교반의 영향 및 Scale-up)

  • Lee, Hak-Su;Lee, Ki-Young;Choi, Sang-Yun;Lee, Shin-Young
    • KSBB Journal
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    • v.26 no.4
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    • pp.357-364
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    • 2011
  • This study was carried out to investigate the effects of agitation, aeration and scale-up on the mycelial growth, exo-polysaccharide (EPS) production, and mycelial morphology in the liquid culture of Ganoderma lucidum. A correlation between roughness and operating variables was also studied to scale-up the liquid culture of G. lucidum in a jar fermenter. When the agitation speed or aeration rate increased, the morphological form was changed from rough pellet to smooth pellet form. Increase of the agitation and aeration reduced the mycelial roughness. On the other hand, in the case of pellet size, it was not affected by aeration. The higher EPS production was obtained at approximately 17% of roughness and mycelial pellet size of 3~5 mm. The morphology at each fermenter was closely correlated with kLa value, and it was found that similarity of morphology would be used as a criteria of scale-up for liquid culture of G. lucidum.

Virulence Reduction and Differing Regulation of Virulence Genes in rpf Mutants of Xanthomonas oryzae pv. oryzae

  • Jeong, Kyu-Sik;Lee, Seung-Eun;Han, Jong-Woo;Yang, Seung-Up;Lee, Byoung-Moo;Noh, Tae-Hwan;Cha, Jae-Soon
    • The Plant Pathology Journal
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    • v.24 no.2
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    • pp.143-151
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    • 2008
  • To define the functions of the rpf genes in Xanthomonas oryzae pv. oryzae (Xoo), which regulates pathogenicity factors in Xanthomonas campestris pv. campestris (Xcc), marker-exchange mutants of each rpf gene were generated. When the mutants were inoculated on a susceptible cultivar, the lesion lengths caused by the rpfB, rpfC, rpfF, and rpfG mutants were significantly smaller than those caused by the wild type, whereas those caused by the rpfA, rpfD, and rpfI mutants were not. Several virulence determinants, including extracellular polysaccharide (EPS) production, xylanase production, and motility, were significantly decreased in the four mutants. However, the cellulase activity in the mutants was unchanged. Complementation of the rpfB and rpfC mutations restored the virulence and the expression of the virulence determinants. Expression analysis of 14 virulence genes revealed that the expression of genes related to EPS production (gumG and gumM), LPS (xanA, xanB, wxoD, and wxoC), phytase (phyA), xylanase (xynB), lipase (lipA), and motility (pitA) were reduced significantly in the mutants rpfB, rpfC, rpfF, and rpfG. In contrast, the expression of genes related to cellulase (eglxob, clsA), cellobiosidase (cbsA), and iron metabolism (fur) was unchanged. The results of this study clearly show that rpfB, rpfC, rpfF, and rpfG are important for the virulence of Xoo KACC10859, and that virulence genes are regulated differently by the Rpfs.